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Abstract

Objectives

This study investigated the Catharanthus roseus phytochemicals by chromatography-mass spectrometry (GC-MS), and in vitro effects on diabetic wound healing based bioassays, molecular docking, and dynamics.

Methods

Fresh C. roseus leaves were extracted using methanol and subjected to preliminary phytochemical screening and GC-MS analysis. The extract was evaluated for antioxidant, anti-inflammatory, antidiabetic, and antimicrobial activities using in vitro assays. Molecular docking and dynamics simulations were performed to analyse interactions of key bioactive compounds with microbial virulence proteins and wound-healing targets.

Results

Phytochemical screening revealed the presence of flavonoids, alkaloids, tannins, saponins, phenols, steroids, carbohydrates, and glycosides. GC-MS identified 63 bioactive compounds, including D-sorbitol, myo-inositol, and palmitic acid etc The extract showed potent antioxidant activity comparable to Vitamin C and significant anti-inflammatory effects via protein denaturation inhibition (89 ± 5.4%, similar to Diclofenac at 91 ± 6.1%). It also inhibited α-amylase and α-glucosidase (69 ± 3.9% and 75 ± 5.4%) respectively, confirming antidiabetic potential. Antimicrobial assays revealed strong inhibition of Streptococcus mutans (MIC: 0.312 mg/mL) and Enterococcus faecalis (0.62 mg/mL). Molecular docking and dynamics revealed that vincristine and vinblastine bind strongly to both microbial virulence proteins and wound-healing targets (VEGF, NF-κB, IL-10), suggesting dual antimicrobial and pro-healing potential. Vincristine showed the highest affinity (–8.9 kcal/mol), indicating stronger therapeutic relevance in diabetic wound repair.

Conclusion

These findings highlight C. roseus bioactive compounds as a promising therapeutic candidate for diabetic wound healing via antioxidant, anti-inflammatory, antidiabetic and antimicrobial activities.

Graphical abstract

This is a visual representation of the abstract.

Keywords

diabetic wound vincristine vinblastine inflammation and angiogenesis

Introduction

Permanent hyperglycaemia is a hallmark of diabetes mellitus, a chronic metabolic condition that can cause serious side effects, such as poor wound healing, neuropathy, nephropathy, and cardiovascular disorders, if untreateted.¹ The incidence of diabetes is on the rise worldwide, both developing and developed countries, where a growing number of cases are linked to high mortality rates and vascular complications.^2,3 About 10% of people with diabetes develop diabetic foot ulcers (DFUs) at some point during their illness, making them a serious concern among these complications.^4,5 People with diabetes type 1 and type 2 are more likely to sustain chronic wounds because of immunological dysregulation, prolonged inflammation, and poor angiogenesis, which all contribute to delayed tissue repair.^6,7

Homeostasis, inflammation, proliferation, and remodelling are all components of the intricately coordinated process of wound healing.⁸ A successful wound healing response requires these stages to occur in the correct sequence and within an appropriate timeframe.⁹ In patients with diabetes there is severe disruption of this process, resulting in prolonged inflammation, oxidative stress, and inadequate collagen deposition, ultimately leading to chronic wounds.¹⁰ Vascular endothelial growth factor (VEGF) plays a crucial role in angiogenesis, promoting new blood vessel formation and tissue repair.¹¹ However, diabetes is associated with reduced VEGF expression, impairing neovascularization and contributing to chronic wound pathology.^12,13 Recent studies highlight that modulating the HIF-1α/VEGF pathway, by inhibiting miR-217 can enhance angiogenesis and accelerate wound healing in diabetic ulcers.¹⁴

Bioactive phytochemicals with antibacterial, anti-inflammatory, wound healing, and antioxidant qualities are abundant in medicinal plants. Plant based sources are account for more than 25% of contemporary medications, demonstrating their potential to provide accessible and affordable treatments.¹⁵ Catharanthus roseus L. (Apocynaceae), also referred to as Vinca rosea or Madagascar periwinkle, is a medicinal subshrub that is well-known for its pharmacological qualities, especially in the treatment of wounds and diabetes.^16,17 There are two flower-coloured variants of this plant, C. roseus (pink) and C. alba (white), identified in Mesopotamian records. C. roseus, which is well-known for their high alkaloid content, especially staphyloxanthin, vinblastine and vincristine, and essential to worldwide medical care.¹⁸ The glossy leaves, pink-to-white flowers, and paired follicle fruits of C. roseus, an evergreen subshrub, grow up to 1 meter tall.¹⁹ The plant's leaf extracts have shown high antibacterial action, especially against Salmonella typhimurium, Pseudomonas aeruginosa, and Staphylococcus aureus, indicating that they may help prevent several infections.²⁰ Compared to C. alba (white flower), an ethanolic root extract of C. roseus showed a higher antioxidant capacity, with benefits growing with concentration.²¹ It is used to treat cancer, particularly childhood leukaemia, because of alkaloids like vincristine and vinblastine.²² This plant also helps heal wounds and treats skin disorders,^23,24 insulin resistance, and high cholesterol levels, tissue regeneration and tissue repair.¹⁰ Also, lowers blood glucose, improves wound healing, and increases tensile strength and hydroxyproline.²⁵ By lowering triglycerides, total cholesterol, VLDL-c, and LDL-c and enhancing the structural characteristics of the heart, liver, and kidney, C. roseus extract from leaves exhibited strong anti-atherosclerotic properties.²⁶ Flower extract greatly increased tissue weight, may be used topically to treat wounds.²⁷ Our study aims to identify the key phytochemicals present in C. roseus extract, and to evaluate its antioxidant, anti-inflammatory, and antimicrobial activities, and to analyse molecular interactions with target proteins involved in antimicrobial and wound healing. The findings may contribute to the development of optimized, plant-based wound care treatments, bridging the gap between traditional medicine and modern therapeutic applications.

Materials and Methods

Chemicals and Reagents

All chemicals and reagents were of analytical grade. Methanol (HPLC grade) was obtained from Merck (USA), and standard phytochemical reagents, including Mayer's, Dragendorff's, Wagner's reagents, ferric chloride, lead acetate, sodium hydroxide, and concentrated sulfuric acid, and Mueller–Hinton agar and broth were from Hi-Media Laboratories (Mumbai, India). DPPH was purchased from Nanjing Duly Biotech Co., Ltd (Nanjing, China). BSA, α-amylase, α-glucosidase, p-nitrophenyl-α-D-glucopyranoside, diclofenac sodium, and ascorbic acid were purchased from Sigma-Aldrich (USA). Streptococcus mutans ATCC-25175, Enterococcus faecalis ATCC-14506, Pseudomonas aeruginosa ATCC- 15442, and Staphylococcus aureus ATCC – 25923, were used as bacterial strains. Protein structures for docking were retrieved from the Protein Data Bank, ligands from PubChem, and analyses were performed using AutoDock Vina, GROMACS, and related bioinformatics tools.

Extract Preparation

Fresh leaves of C. roseus were collected, washed, and shade-dried. The dried material was powdered and subjected to extraction using methanol by maceration extraction. The extract was concentrated under reduced pressure using a rotary evaporator and stored at −20 °C for further analysis (Figure 1).

Figure 1.

Sequential process of Catharanthus roseus plant extraction.

Phytochemical Screening

Phytochemical investigations were performed to verify the existence of different chemicals in plant extracts. Whereas flavonoids created a green precipitation with 10% ferric chloride, alkaloids produced a reddish-brown precipitate with Wagner's reagent. 5% ferric chloride caused phenols to become blue-black or dark green.²⁸ A brown ring at the contact was used to identify glycosides using glacial acetic acid, FeCl₃, and H₂SO₄.^29,30 With chloroform and H₂SO₄, terpenoids and steroids produced a reddish-brown hue.³¹ When combined with ferric chloride, tannins were seen as either blue (gallic) or greenish-black.³² After shaking with water, saponins created a 1 cm layer of foam.^29,33 Proteins were coloured purple with ninhydrin, whereas carbohydrates created a violet ring with sulphuric acid and α-naphthol.^34–36

GC-MS Analysis

GC/MS analysis was performed using an Agilent 5977B system equipped with a DB5-MS column (29 m × 0.25 mm × 0.25 µm) and a 10 m DuraGard column. The oven temperature was ramped from 60 °C to 325 °Cover 37.5 min, with helium as the carrier gas at a constant flow rate of 1 mL/min. Sample injection was performed at 250 °C under both split less conditions (helium purge 10.5 mL/min for 1 min) and split mode (1:10, split flow 10.3 mL/min). The MSD was set to a source temperature of 230 °C and a quadrupole temperature of 150 °C, scanning a mass range of 50-600 u after a 5.90 min solvent delay. Retention times were standardized using d27-myristic acid with Agilent's RTL system for consistency. Metabolite identification was carried out using the NIST metabolomics library.³⁷

Antioxidant Activity-DPPH Radical Scavenging Assay

The scavenging activity of the DPPH radical was evaluated according to the method described by Rajesh and Natvar (2011).³⁸ Briefly, DPPH solution was prepared in 3.3 mL of methanol and protected from light using aluminium foil. Different concentration of the extracts were individually mixed with 150 µL of the DPPH solution, and the final volume was adjusted to 3 mL with methanol. After incubation for 15 min at room temperature in the dark, the absorbance was measured at 517 nm using a Shimadzu UV-1601 UV–visible spectrophotometer, and methanol was used as the blank. The following formula was used to calculated inhibition percentage:

\begin{aligned} DPPHRadicalsScavenged (%) \\ = (Control OD - Sample OD / Control OD) \times 100 \end{aligned}

Anti-inflammatory Activity by Protein Denaturation Assay

A volume of 1 ml of extracts or of diclofenac sodium at different concentrations 0.025, 0.05, 0.1, 0.15 and 0.2 mg/mL was homogenised with 1 ml of aqueous solution of bovine serum albumin (5%) and incubated at 27 °C for 15 min. The mixture of distilled water and BSA constituted the control tube. Denaturation of the proteins was caused by placing the mixture in a water bath for 10 min at 70 °C. The mixture was cooled at room temprature, and the activity each mixture was measured at 660 nm.³⁹ Each test was done in three times. The following formula was used to calculated inhibition percentage:

\begin{aligned} PercentageInhibition \\ = (Control OD – Sample OD / Control OD) \times 100 \end{aligned}

Anti-diabetic Activity: α-Amylase & α-Glucosidase Inhibitory Activity

The α-Amylase inhibitory activity was determined using the 3,5-dinitrosalicylic acid (DNS) method. Briefly, a reaction mixture containing phosphate buffer, α-amylase enzyme, and varying concentrations of the plant extract were pre-incubated under controlled conditions. The reaction was initiated by the addition of starch substrate and incubated to allow enzymatic hydrolysis. The enzymatic reaction was terminated by the addition of DNS reagent, followed by heating to develop colour. After cooling, the optical density was measured spectrophotometrically, and extent of α-amylase inhibition was calculated. The α-Glucosidase inhibitory activity was evaluated by pre-incubating a mixture of phosphate buffer, α-glucosidase enzyme, and plant extract. The reaction was initiated by adding p-nitrophenyl-α-D-glucopyranoside (p-NPG) as the substrate and incubated for a defined period. The reaction was terminated by the addition of sodium bicarbonate, and the absorbance of the released p-nitrophenol was measured spectrophotometrically. Acarbose was used as the standard reference inhibitor in both assays, and percentage inhibition was calculated in comparison with the control.⁴⁰

Antimicrobial Activity: Determination of Minimum Inhibitory Concentration (MIC)

The minimum inhibitory concentration (MIC) of C. roseus was determined using the micro dilution technique at concentrations ranging from 10 mg/mL to 0.019 mg/mL and 23,5-triphenyltetrazolium chloride (TTC) was used as the viability indicator. Viable bacterial cells reduce the colourless TTC to a red-coloured insoluble formazan. After incubation, wells showing no colour development indicated complete inhibition of bacterial growth and were considered as MIC.

Molecular Docking

Protein–ligand interactions were investigated using molecular docking analysis. Initially, the three-dimensional structure of the target protein was retrieved from the Protein Data Bank (PDB) and prepared for docking analysis. Ligand preparation (staphyloxanthin (STX), vinblastine (VBL), and vincristine (VCR)) involved obtaining the three-dimensional structures either from the PubChem database or generating them using appropriate computational tools. During protein preparation, only polar hydrogen atoms were added to facilitate accurate intermolecular interactions. For ligand preparation, Gasteiger charges were assigned and non-polar hydrogen atoms were added. Rotatable bonds within the ligand molecules were identified and optimized, after which each prepared ligand was individually uploaded into the docking software. Molecular docking simulations were performed using AutoDock version 1.5.6, and the resulting protein–ligand interactions were analyzed and visualized in both two- and three-dimensional formats using BIOVIA Discovery Studio.⁴²

Molecular Dynamics

Molecular dynamics (MD) simulations were performed using the GROMACS software package, with additional computational support provided by the WebGRO server (https://simlab.uams.edu/). The docked protein–ligand complex was separated into receptor (protein) and ligand components to facilitate system preparation. Protein topology files were generated in GROMACS using the AMBER99SB-ILDN force field, while ligand topology parameters were obtained using the PRODRG server. Subsequently, the individual topologies were combined to reconstruct the protein–ligand complex for simulation. The complex was placed in a dodecahedral simulation box with a minimum distance of 1.0 nm between the complex and the box boundaries. The system was solvated using the TIP3P water model, and electrostatic neutrality was achieved by the addition of 0.15 M NaCl. MD simulations were carried out for 10 ns under standard conditions using the GROMACS command-line interface on an Ubuntu operating system. Post-simulation analyses were performed using the generated trajectory (XTC) files to evaluate the root mean square deviation (RMSD) and root mean square fluctuation (RMSF). These analyses were used to assess the conformational stability and dynamic behaviour of the protein–ligand complex throughout the simulation period, thereby confirming the structural integrity of the complex under dynamic conditions.

Statistical Analysis

The data were statistically assessed using one-way analysis of variance (ANOVA) and Duncan's multiple comparison test using computerized software (SPSS 7.5 using Windows student version) to determine the significance of difference between different groups. The experiment was conducted with a significance criterion of p < 0.05.

Results

Phytochemical Analysis

The phytochemical analysis of C. roseus shows the presence of several bioactive compounds that aid diabetic wound healing (Figure 2 and Table 1). Flavonoids, saponins, tannins, phenols, and carbohydrates are abundant. Alkaloids, steroids, and glycosides are moderately present. Terpenoids and proteins are either absent or present in negligible amounts. These compounds are known to work together to enhance healing by reducing oxidative stress, fighting infections, and supporting tissue repair.

Figure 2.

Various types of phytochemicals found in the methanol extract of C. roseus. 1. Proteins, 2. Carbohydrates, 3. Flavonoids, 4. Phenols, 5. Terpenoids, 6. Glycosides, 7. Alkaloids, 8. Saponins, 9. Tannins and 10. Steroids.

Table 1.

Preliminary Phytochemical Study of C. roseus Methanol Extracts.

S.No	Phytochemicals	Inference
1	Proteins	-
2	Carbohydrates	+++
3	Flavonoids	+++
4	Phenols	+++
5	Terpenoids	-
6	Glycosides	++
7	Alkaloids	++
8	Saponins	+++
9	Tannins	+++
10	Steroids	++

Phytochemicals were represented by the symbols “+, ++, and +++” to indicate their presence low, moderate, and high, whereas “-“denoted their absence.

GC-MS Analysis of a Methanolic Extract of Catharanthus Roseus

GC–MS analysis of the methanolic extract of Catharanthus roseus revealed the presence of multiple bioactive constituents. The identification of compounds was performed by comparing the obtained mass spectra with those available in the NIST mass spectral library. Several major constituents were detected, including D-fructose, 1,3,4,5,6-pentakis-O-(trimethylsilyl)-, O-methyloxime, D-galactose, D-(+)-talose, D-sorbitol, myo-inositol, and palmitic acid. The reported match or similarity indices for these compounds were high hit score. Our preliminary phytochemical test showed the presence of alkaloids and many literature explored that staphyloxanthin, vinblastine and vincristine were abundant in C. roseus, however our GC–MS conditions not amenable to these phytochemicals. The identified compounds are known to exhibit diverse biological activities relevant to diabetic wound healing. Sugars and sugar alcohols, such as fructose, galactose, sorbitol, talose, and myo-inositol, play important roles in cellular energy metabolism, osmotic balance, and tissue hydration, thereby supporting wound repair processes. Palmitic acid contributes to membrane integrity and may influence inflammatory signaling. Collectively, these bioactive constituents suggest that the methanolic extract of C. roseus possesses significant antioxidant and anti-inflammatory potential. The extract may promote diabetic wound healing by reducing NF-κB-mediated inflammatory responses, enhancing angiogenesis through VEGF regulation, and facilitating tissue regeneration. The GC-MS chromatogram and bioactive compounds present in the extracts were summarized in Figure 3 and Table 2.

Figure 3.

GC-MS chromatogram of methanolic extract of the C. roseus.

Table 2.

Bioactive Compounds Identified from GC-MS Analysis of Methanolic Extract of C. roseus.

RT (min)	Compound Name	Formula	Area	Hit Score
6.1142	Benzene, 1,2-dichloro-	C6H4Cl2	5466443	93.5
6.3810	Tetrasiloxane, decamethyl-	C10H30O3Si4	870621	85.8
6.6917	Lactic Acid, 2TMS derivative	C9H22O3Si2	2152034	97.5
7.7068	3-Pyridinol, TMS derivative	C8H13NOSi	271901	88.7
7.6691	3-Furoic acid, TMS derivative	C8H12O3Si	63288	90.5
8.1712	Pentasiloxane, dodecamethyl-	C12H36O4Si5	819517	82.1
8.3794	5-Hydroxy-2-methylpyridine, TMS derivative	C9H15NOSi	308739	84.5
9.1678	Benzoic Acid, TMS derivative	C10H14O2Si	184092	89.4
9.2806	Bisphenol A monomethyl ether, TMS derivative	C19H26O2Si	65284	80.4
9.6009	Glycerol, 3TMS derivative	C12H32O3Si3	54672501	84.4
9.7806	Niacin, TMS derivative	C9H13NO2Si	609517	92.4
9.8405	Valylvaline, N, N’dimethyl-N'-ethoxycarbonyl-, undecyl ester	C26H50N2O5	400984	83.3
9.8498	12,3-Butanetriol, 3TMS derivative	C13H34O3Si3	4587099	96.9
9.9566	2-Butenedioic acid, (Z)-, 2TMS derivative	C10H20O4Si2	2629510	97.3
10.0517	Butanedioic acid, 2TMS derivative	C10H22O4Si2	3192133	96.8
10.3625	Glyceric acid, 3TMS derivative	C12H30O4Si3	7014924	94.0
10.4685	2-Butenedioic acid, (E)-, 2TMS derivative	C10H20O4Si2	1937815	90.1
11.0380	Etoxeridine	C18H27NO4	25085	85.6
11.6628	Butanoic acid, 3,4 bis[(trimethylsilyl)oxy]-, trimethylsilyl ester	C13H32O4Si3	2199693	89.6
12.4086	Malic acid, 3TMS derivative	C13H30O5Si3	40764939	98.3
12.6203	L-Threitol, 4TMS derivative	C16H42O4Si4	95723603	97.4
12.7185	L-Threitol, 4TMS derivative	C16H42O4Si4	87564186	97.0
12.7292	Etoxeridine	C18H27NO4	56792	80.2
12.7969	L-5-Oxoproline, 2TMS derivative	C11H23NO3Si2	13881685	92.0
12.8781	4-Aminobutanoic acid, 3TMS derivative	C13H33NO2Si3	8477480	94.7
13.1490	23,4-Trihydroxybutyric acid tetrakis(trimethylsilyl) deriv., (, (R,R)-)	C16H40O5Si4	2998307	90.6
13.3518	23,4-Trihydroxybutyric acid tetrakis(trimethylsilyl) deriv., (, (R,R)-)	C16H40O5Si4	12836057	96.4
13.5881	Diethyl Phthalate	C12H14O4	231456	92.6
13.9775	4-Hydroxybenzoic acid, 2TMS derivative	C13H22O3Si2	5251736	92.1
14.0631	Pentitol, 3-desoxy-tetrakis-O-(trimethylsilyl)-	C17H44O4Si4	1045365	83.6
14.1182	4-Hydroxybenzeneacetic acid, 2TMS derivative	C14H24O3Si2	3859693	92.5
14.1252	Xylonic acid, 23,4-tris-O-(trimethylsilyl)-,.delta.- lactone, D-	C14H32O5Si3	3900025	80.2
14.4560	34,5-Trihydroxypentanoic acid, tetrakis(trimethylsilyl)-	C17H42O5Si4	4820741	90.9
14.5828	D-Arabinose, tetrakis(trimethylsilyl) ether, ethyloxime (isomer 2)	C19H47NO5Si4	5836795	96.7
14.6056	2,6-Dimethoxyhydroquinone, 2O-TMS	C14H26O4Si2	245912	81.0
14.9756	Xylitol, 5TMS derivative	C20H52O5Si5	18638118	90.7
15.1020	D-(+)-Arabitol, 5TMS derivative	C20H52O5Si5	9810260	95.1
15.2485	D-(+)-Arabitol, 5TMS derivative	C20H52O5Si5	91356093	97.4
15.2959	Adonitol, 5TMS derivative	C20H52O5Si5	5788680	94.1
15.5234	Vanillic Acid, 2TMS derivative	C14H24O4Si2	3527483	95.6
15.6516	Phosphoric acid, bis(trimethylsilyl) 2,3-bis[(trimethylsilyl)oxy]propyl ester	C15H41O6PSi4	835975	81.4
15.9179	2-Keto-l-gluconic acid, penta(O-trimethylsilyl)-	C21H50O7Si5	18400352	84.3
16.0847	Shikimic acid, 4TMS derivative	C19H42O5Si4	16698206	95.3
16.1563	Protocatechoic acid, 3TMS derivative	C16H30O4Si3	21710018	88.9
16.3049	3-Deoxyhexitol, 5TMS derivative	C21H54O5Si5	8165862	89.6
16.4005	3-Deoxyhexitol, 5TMS derivative	C21H54O5Si5	43389223	92.4
16.4797	D-Pinitol, pentakis(trimethylsilyl) ether	C22H54O6Si5	79020813	95.2
16.7637	Semioxamazide	C2H5N3O2	3460783	82.8
16.8155	Silane, diethylheptadecyloxy(2-methylbutoxy)-	C26H56O2Si	188437	80.1
16.9004	D-Fructose, 1,3,4,5,6-pentakis-O-(trimethylsilyl)-, Omethyloxime	C22H55NO6Si5	233180953	91.6
17.0286	Disiloxane, 1,3-bis(1,1-dimethylethyl)-1,1,3,3- tetramethyl-	C12H30OSi2	49862689	80.5
17.0293	d-Galactose, 2,3,4,5,6-pentakis-O-(trimethylsilyl)-, omethyloxyme, (1E)-	C22H55NO6Si5	123246992	82.5
17.1100	D-Allose, pentakis(trimethylsilyl) ether, methyloxime (syn)	C22H55NO6Si5	47498282	81.2
17.1587	D-(+)-Talose, pentakis(trimethylsilyl) ether, methyloxime (syn)	C22H55NO6Si5	60708221	92.7
17.5200	D-Sorbitol, 6TMS derivative	C24H62O6Si6	70948818	96.6
17.5937	D-Sorbitol, 6TMS derivative	C24H62O6Si6	97703493	94.9
17.6171	Gallic acid, 4TMS derivative	C19H38O5Si4	13111098	87.2
18.2056	D-Gluconic acid, 6TMS derivative	C24H60O7Si6	9432481	88.6
18.2778	Palmitic Acid, TMS derivative	C19H40O2Si	53875912	98.4
18.4596	Silane,dimethyl(docosyloxy)undecyloxy-	C35H74O2Si	108420	81.2
18.7765	Ferulic acid, 2TMS derivative	C16H26O4Si2	1394820	86.4
18.9236	N-Acetyl-D-glucosamine, tetrakis(trimethylsilyl) ether, methyloxime (syn)	C21H50N2O6Si 4	3662866	86.4
19.0124	Myo-Inositol, 6TMS derivative	C24H60O6Si6	114190751	95.3
19.2016	Caffeic acid, 3TMS derivative	C18H32O4Si3	3581737	95.0
19.2491	D-Allose, pentakis(trimethylsilyl) ether, ethyloxime (isomer 1)	C23H57NO6Si5	9811083	90.3
19.2500	D-Altrose, pentakis(trimethylsilyl) ether, ethyloxime (isomer 2)	C23H57NO6Si5	10278307	89.4
19.3351	1-(2-(3-Cyclohexenyl) ethyl) silatrane	C14H25NO3Si	834384	86.2
19.3657	9-Octadecenoic acid, ethyl ester	C20H38O2	1562205	84.4
19.3657	(E)-9-Octadecenoic acid ethyl ester	C20H38O2	1630646	84.3
19.3841	Galactose oxime, 6TMS derivative	C24H61NO6Si6	1698885	86.6
19.3844	D-Allose, oxime (isomer 1), 6TMS derivative	C24H61NO6Si6	1844316	85.2
19.5020	D-Allose, oxime (isomer 2), 6TMS derivative	C24H61NO6Si6	2783007	87.0
19.8327	Oleic Acid, (Z)-, TMS derivative	C21H42O2Si	5857361	95.7
20.0308	Stearic acid, TMS derivative	C21H44O2Si	11121353	97.6
20.1966	D-Sorbitol, 6TMS derivative	C24H62O6Si6	6394703	89.4
20.1967	D-Mannitol, 6TMS derivative	C24H62O6Si6	5701926	89.6
20.3624	1-Phenyl-1,2-ethanediol, 2TMS derivative	C14H26O2Si2	46362416	82.2
20.9752	Glyceryl-glycoside TMS ether	C27H66O8Si6	11222950	93.9
21.3461	Silane, diethyldodecyloxypentyloxy-	C21H46O2S	259311	80.7
21.6627	Arachidic acid, TMS derivative	C23H48O2Si	1537250	83.5
22.8796	1-Monopalmitin, 2TMS derivative	C25H54O4Si2	1474730	84.5
23.1536	Phthalic acid, di(3-ethylphenyl) ester	C24H22O4	1217875	81.1
24.2278	Sucrose, 8TMS derivative	C36H86O11Si8	2964740	87.6
24.2741	Chrysophanol, 2TMS derivative	C21H26O4Si2	38316	85.3
24.2900	Silane, diethylheptadecyloxy(2-methylbutoxy)-	C26H56O2Si	113437	88.0
24.4367	Lactose, 8TMS derivative	C36H86O11Si8	75692003	96.6
25.2491	Methyl 2,6-dihydroxybenzoate, 2TMS derivative	C14H24O4Si2	58835	82.0
25.9217	Silane, dimethyl (4-(2-phenylprop-2- yl) phenoxy) tridecyloxy-	C30H48O2Si	6946	82.0
26.0492	Lactose, 8TMS derivative	C36H86O11Si8	10057813	80.2
26.8467	Chlorogenic acid (6TMS)	C34H66O9Si6	935163	81.7
29.8522	Sucrose, 8TMS derivative	C36H86O11Si8	3031230	84.2
31.9984	Silane, diethyl(pentafluorobenzyloxy)tetradecycloxy-	C36H64O3Si2	8022808	91.9

Effects of Antioxidant DPPH Activity of Extract

The DPPH radical scavenging assay demonstrated a marked, concentration-dependent antioxidant activity of the C. roseus extract (Figure 4). The percentage of inhibition increased from approximately 16% at the lowest tested concentration to about 79% at 0.2 mg/ml, indicating a strong free radical–quenching ability. This activity was comparable to the standard antioxidant Vitamin C, which showed 86% inhibition at the same concentration. The close similarity in scavenging efficiency at higher doses highlights the potent antioxidant potential of the extract, suggesting its effectiveness in reducing oxidative stress, a key factor impairing tissue repair and diabetic wound healing (Table S.T1).

Figure 4.

Antioxidant activity of the extract assessed by the DPPH radical scavenging assay. The extract exhibited strong free-radical scavenging activity comparable to the standard, as determined by percentage inhibition of DPPH. Data are expressed as mean ± SD of triplicates, and statistical significance was considered at p < 0.05, and a, b, c & d represents significance difference among different concentrations.

Effects of Anti-Inflammatory Activity of Extract

The anti-inflammatory activity assessed by the protein denaturation assay showed a strong concentration-dependent inhibitory effect of the extract (Figure 5). Inhibition increased steadily from approximately 22% at 0.025 mg/ml to about 89% at 0.2 mg/ml, demonstrating effective prevention of protein denaturation. This response was comparable to the standard diclofenac, which exhibited inhibition ranging from ∼28% at the lowest concentration to 91% at 0.2 mg/ml. At higher concentrations (0.1-0.2 mg/ml), the extract showed no significant difference from the standard drug, indicating potent anti-inflammatory efficacy. These findings suggest that the extract possesses strong anti-inflammatory potential, supporting its therapeutic relevance in managing inflammation during diabetic wound healing (Table S.T2).

Figure 5.

Protein denaturation assay showing the anti-inflammatory activity of C. roseus extract at various concentrations. Values are expressed as mean ± SD of triplicates. Statistical significance was determined at p < 0.05, and a, b, c & d represents significance difference among different concentrations.

Effects of α-Amylase Inhibition Activity of Extract

The α-amylase inhibitory assay revealed that the extract exhibited a clear concentration-dependent antidiabetic effect (Figure 6). The percentage inhibition increased from approximately 19% at 0.1 mg/ml to about 69% at 0.5 mg/ml, indicating its ability to suppress carbohydrate-digesting enzyme activity. Although the inhibitory effect was slightly lower than that of the standard drug acarbose, which showed inhibition ranging from ∼20% to 80% over the same concentration range, the extract demonstrated statistically significant activity at higher doses. These findings suggest that the extract can effectively slow carbohydrate digestion and postprandial glucose release, supporting its potential role in glycemic control and the management of diabetes-associated wound healing (Table S.T3).

Figure 6.

α-amylase inhibitory activity of C. roseus extract at various concentrations. Values are expressed as mean ± SD of triplicates. Statistical significance was determined at p < 0.05, and a, b, & c represents significance difference among different concentrations.

Effects of α-Glucosidase Inhibition Activity of Extract

The α-glucosidase inhibition assay demonstrated a significant, dose-dependent antidiabetic activity of the extract (Figure 7). The extract inhibited α-glucosidase from approximately 16% at 0.1 mg/ml to about 75% at 0.5 mg/ml, indicating effective suppression of intestinal carbohydrate-digesting enzymes. Although its activity was marginally lower than the standard drug acarbose, which showed inhibition ranging from ∼18% to 79% over the same concentration range, the extract exhibited comparable efficacy at higher doses. These findings suggest that the extract can effectively delay carbohydrate breakdown and glucose absorption, contributing to improved glycemic control and supporting its potential role in the management of diabetes-associated wound healing (Table S.T4).

Figure 7.

α-glucosidase inhibitory activity of C. roseus extract at various concentrations. Values are expressed as mean ± SD of triplicates. Statistical significance was determined at p < 0.05, and a, b, & c represents significance difference among different concentrations.

Minimum Inhibitory Concentration

The antimicrobial effects of C. roseus extract demonstrated in table 3. Strong antibacterial activity was observed against S. mutans and E. faecalis at higher concentrations (10 to 0.62 mg/mL). At 0.312 mg/mL, S. mutans exhibit no growth, while E. faecalis growth was observed in this concentration. At lower concentrations (0.156 mg/mL and below), both bacterial strains exhibit growth, showing the extract's loss of antimicrobial activity due to less concentration of bioactive compounds. The extract had no effect on P. aeruginosa and S. aureus. The minimum inhibitory concentrations were 0.312 mg/mL for S. mutans and 0.62 mg/mL for E. faecalis.

Table 3.

Anti-bacterial activity of methanol extracts of C. roseus. Concentrations (mg/mL), (-) no Bacterial Growth and (+) Bacterial Growth.

	Concentration of C. Roseus	Growth Measured
S. No	(mg/mL)	S. mutans	E. faecalis	S. aureus	P. aeruginosa
1	10	−	−	+	+
2	5	−	−	+	+
3	2.5	−	−	+	+
4	1.25	−	−	+	+
5	0.62	−	−	+	+
6	0.312	−	+	+	+
7	0.156	+	+	+	+
8	0.078	+	+	+	+
9	0.039	+	+	+	+
10	0.019	+	+	+	+

In-silico Analysis - Docking Studies with Microbial Proteins

According to docking studies, the microbial enzymes glycosyltransferase, glucosidase, mannosidase, elastase, and serine protease are firmly bounded by vincristine and vinblastine with binding energies ranging from −7.4 to −9.7 kcal/mol. These interactions imply that protease-mediated tissue injury and bacterial glucose metabolism are inhibited. The chemicals improve VEGF-mediated angiogenesis and indirectly modify TNF-α by decreasing microbial pathogenicity and excessive protease activity. Therefore, under diabetic circumstances, C. roseus has both antibacterial and wound-healing properties (Table 4).

Table 4.

Vinblastine and Vincristine, Bioactive Compounds of C. roseus, Molecularly Target with Bacterial Proteins.

Docking Studies with Wound Healing Proteins

STX, VBL, and VCR exhibited notable binding affinities toward key proteins involved in angiogenesis, inflammation, insulin signalling, and wound repair, including AKT, VEGF, EGF, eNOS, FGF, HIF-1α, IL-1β, IL-10, IR, IRS-1, MCP-1, MMP-9, NF-κB, PDGF, SDF-1α, SOD, TGF-β1, and TNF-α. STX showed moderate binding energies ranging from −5.1 to −7.3 kcal/mol and formed one to three hydrogen bonds per protein, primarily involving ARG, LYS, and GLN residues, contributing to stable protein–ligand interactions. In comparison, VBL demonstrated stronger binding affinities (−5.6 to −8.7 kcal/mol) and formed conventional hydrogen bonds with proteins such as IL-10, AKT, and HIF-1α, involving key residues including LYS, ASP, GLU, and TYR. VCR exhibited the highest interaction stability, forming up to six hydrogen bonds with targets such as TGF-β1, EGF, and Nrf2, mediated by residues including ASN, SER, GLN, and ARG. Collectively, these results indicates that bioactive compounds from C. roseus, particularly vinblastine and vincristine, possess strong multi-target binding potential, supporting their therapeutic relevance in modulating inflammatory, angiogenic, and insulin-related pathways associated with diabetic wound healing (Table 5 and Figures S.5–7).

Table 5.

Binding Affinity of the Protein-Ligand Docking Complex.

	Bonding Energy (Kcal/mol)			Number of Hydrogen Bonds			Residues
Proteins	Drugs			Drugs			Drugs
	VBL	VCR	STX	VBL	VCR	STX	VBL	VCR	STX
VEGF	−5.6	−6.2	−5.9	3	1	-	LYS C:313
							CYS C:267
							ASN C:268
TNF-α	−5.8	−6.1	−5.2	1	-	2	GLN A:114		HIS B:119
TYR B:166
TGF-β – 1	−7.1	−8	−7.2	2	3	4	ASP A:382
TYR A:306	GLN A:270
ASN A:275
MET A:110	ASN A:403
LYS A:410
PRO A:380
GLN A:46
SDF-1 α	−5.9	−6.2	−5.2	3	-	1	LEU A:57
CYS A:55
THR A:52	-	GLN A:69
PDGF	−5.9	−6.2	−5.2	1	-	-	ALA A:91	-	-
Nrf2	−8.4	−8.5	−6.7	1	2	2	GLY A:372	THR A:560
VAL A:608	VAL A:420
ARG A:326
SOD	−6.3	−7	−5.9	2	1	2	GLU A:191
ASN A:188	HIS A:163	HIS A:2
ARG A:192

NF-kB	−7.4	−7.6	−6.1	2	2	1	ASP A:610
LYS A: 637
LEU A:606
	ALA A:604
LEU A:608	ASN A:614
MMP-9	−7.2	−7.9	−7.3	2	1	2	GLU A:130
SER A:129	GLN A:90	LYS A:656
GLN A:593
MCP-1	−6.5	−6.8	−5.1	3	2	2	MET D:52
PHE D:49
LYA D:73	LYS D:208
PRO D:206	LYS D:208
ASP D:201
IRS-1	−6.8	−6.6	−5.9	1	2	3	GLU A:45	LYS A;161
ARG A:75	GLU A:91
ARG A:62
GLN A:165
IR	−7.3	−6.2	−6.8	2	3	2	GLN A:111
SER A:1270	HIS A:1057
ARG A:1061
LYS A:1020
	ASN A:1137
SER A:1006
IL-10	−8.7	−8.9	−7.1	1	-	1	PHE A:129	-	ARG A:45
IL-1β	−7.2	−8.2	−5.4	1	1	3	ARG A:75	GLU A:99	GLN A:37
LYS A:73
GLU A:99
HIF-1α	−7.8	−7.7	−6.6	3	6	3	ARG A:120
TYR A:93
GLN A:147	TYR A:102
TYR A:93
SER A:91
THR A:149
GLN A:148
GLN A:147	GLU A:122
THR A:183
SER A:184
FGF	−6.8	−7.3	−5.8	2	3	3	SER A:50
TYR A:131	ARG A:12
SER A:50
ASP A:129	LEU A:162
SER A:163
SER A:50
EGF	−7.9	−8.1	−6.2	3	3	2	ASP A:502
THR A:501
TYR A:467	ASN A:540
ASN A:524
THR A:381	ASN A:113
TYR A:467
eNOS	−7.9	−8.4	−6.7	3	2	1	GLY A:463
ALA A:86
ALA A:88	ARG A:365
ARG A:372	ASP A:287
AKT	−7.7	−7.5	−6.4	5	-	1	ASP A:292
LYS A:179
PHE A:161
THR A:160
LYS A:276	-	HIS A:220

Molecular Dynamics

Molecular dynamics (MD) simulations conducted for 10 ns further confirmed the stability of the identified bioactive compounds against key diabetic wound-related targets. The MMP-9–STX complex exhibited a stable RMSD trajectory (∼0.14-0.17 nm) throughout the simulation, with minimal deviations, suggesting strong conformational stability. Similarly, the Serine Protease–VBL complex displayed a consistently low RMSD (∼0.11-0.13 nm), further validating the structural integrity of the binding interaction. RMSF analysis supported these observations, revealing only minor fluctuations in most residues (<0.2 nm), with higher flexibility limited to loop and terminal regions. In MMP-9, local fluctuations reached up to 2.5 nm in selective segments, yet the binding site residues remained stably anchored, while the Serine Protease complex showed negligible flexibility with fluctuations remaining below 0.3 nm. These findings collectively indicate that C. roseus phytoconstituents form dynamically stable interactions with diabetic wound-associated targets, modulating TNF-α/VEGF signalling to enhance wound repair mechanisms and suppress pathogen-associated proteolytic activity (Figure 8).

Figure 8.

RMSD and RMSF plots of MMP-9 with STX and serine protease with VBL over 10 ns md simulations, showing stable protein–ligand interactions. Minor residue fluctuations were confined to loop regions, confirming complex stability in regulating TNF-α/VEGF-mediated diabetic wound healing.

Discussion

Wound healing is a complex and dynamic process that involves the interplay of various cell types, signalling molecules, and extracellular matrix components. In diabetic conditions, this repair mechanism is markedly impaired due to chronic inflammation, oxidative stress, and poor angiogenesis, often resulting in non-healing ulcers and amputations.⁴¹ Although herbal ointments have shown promise in reducing amputation rates in diabetic wounds,⁴² systematic research is needed to validate their efficacy and establish molecular mechanisms. In this context, the present study investigated the therapeutic potential of C. roseus in diabetic wound healing, comparing its bioactive properties with previous reports. Phytochemical profiling of C. roseus extracts revealed the presence of alkaloids, flavonoids, tannins, saponins, carbohydrates, phenols, steroids, and glycosides, compounds well-known for their pharmacological relevance (Figure 2 and Table 1). These findings align with earlier studies,^43–46 which documented more than 400 alkaloids in this plant, including vinblastine and vincristine, with established pharmacological and anticancer effects. Previous studies have demonstrated that C. roseus exhibits part-specific phytochemical enrichment with relevance to metabolic and angiogenic regulation. Leaf extracts predominantly contain terpenoids such as phytol, while flower extracts are enriched with fatty acid esters, including D-sorbitol, myo-inositol, and palmitic acid.⁴⁷ These metabolites have been associated with antioxidant, anti-inflammatory, and metabolic regulatory effects, which are critical in diabetes-associated vascular impairment. Furthermore, Amna Parveen et al (2019)⁴⁸ reported that natural products can interact with VEGF-promoting factors through multiple signalling pathways, supporting the role of plant-derived compounds in modulating angiogenesis-linked metabolic dysfunction. Phenolic and alkaloid constituents such as vincristine and vinblastine have also been reported to influence angiogenesis and inflammatory signaling,⁴⁹ processes closely linked to VEGF-mediated vascular repair in diabetic conditions. Previous literature reports part-specific phytochemical enrichment in C. roseus, with leaf extracts rich in terpenoids such as phytol (57.47%) and flower extracts enriched in fatty acid esters, including palmitic acid (34.65%) and methyl 9-octadecanoate (29.88%).^50,51 Methanolic leaf extract profiling further identified D-sorbitol, myo-inositol, and palmitic acid, along with phenolics and alkaloids such as staphyloxanthin, vincristine and vinblastine. The presence of these VEGF-associated and metabolically relevant compounds supports the conclusion that C. roseus may exert antidiabetic effects through regulation of VEGF-linked signalling pathways.

Oxidative stress is one of the major contributors to delayed wound healing in diabetes. C. roseus extracts exhibited strong antioxidant capacity in a dose-dependent manner. Previous research reported 55% DPPH inhibition for leaf extract in ethanolic solvent.^50,52,53 Unlike prior studies, our research provides a direct comparison with vitamin C, underscoring the plant's potential for diabetic wound care. Extracts of C. roseus leaves and flowers have demonstrated dose-dependent hypoglycaemic effects, enhancing liver glucose utilization.^54–56 These extracts showed significant hypoglycaemic, hypolipidemic, and anti-hyperglycaemic properties by improving glucose metabolism, enzymatic activity, insulin levels, and sensitivity in diabetic rats.^53,55 The DPPH radical scavenging activity increased from 16 ± 0.9% to 79 ± 4.6%, values comparable to Vitamin C (86 ± 6.2%) (Figure 4). These results are consistent with earlier reports,⁵⁷ which demonstrated similar free-radical scavenging activity. In addition, C. roseus extracts improved antioxidant enzyme activity (CAT, SOD, GPx, GST) and enhanced glycogen storage, highlighting its role in mitigating oxidative stress and supporting metabolic balance.

The C. roseus extracts exhibited concentration-dependent anti-inflammatory and protein denaturation inhibition activities, with the composite showing slightly lower effectiveness than diclofenac.^58,59 In the present study, the anti-inflammatory potential of C. roseus was further confirmed through protein denaturation inhibition assays. The extract demonstrated a concentration-dependent effect, with inhibition values ranging from 22 ± 1.7% to 89 ± 5.4%. At higher concentrations (0.15-0.2 mg/ml), activity was comparable to diclofenac, a standard anti-inflammatory drug (Figure 5). These findings suggest that C. roseus effectively reduces inflammation, a key pathological barrier in diabetic wound repair. In addition, moderate inhibitory activity exhibited against both α-glucosidase and α-amylase enzymes. At a concentration of 0.5 mg/ml, α-amylase inhibition reached 69 ± 3.9%, suggesting their potential in controlling postprandial glucose spikes, while α-glucosidase inhibition increased to 75 ± 5.4%, indicating a role in reducing glucose absorption in the small intestine (Figure 6 & 7). These findings highlight the plant's potential in managing blood sugar levels. The antimicrobial activity of C. roseus was also evident, with methanolic extracts showing strong inhibitory activity against E. faecalis (MIC: 0.62 mg/mL) and S. mutans (MIC: 0.312 mg/mL), however no significant effects was observed against P. aeruginosa and S. aureus (Table 3). These results are consistent with earlier research^57,60 that showed C. roseus had antibacterial activity against B. subtilis, E. coli, and S. typhi. However, at 20%–40% concentrations, moderate inhibition against S. aureus and E. coli, as reported by Kabesh et al (2015).⁶¹ Importantly, our study demonstrated that the antibacterial effects of C. roseus are concentration-dependent, adding clarity to earlier findings.

Many studies reported that staphyloxanthin, vincristine and vinblastine compounds abundant in C. roseus, thus we performed docking analysis with these compounds. The results demonstrated strong hydrogen bond interactions with active site residues and high binding affinity to the target proteins. Compounds from C. roseus, such as vinblastine, ajmalicine, vindolinine, and reserpine, have the potential to control hypertension and diabetes by modifying HPA activity.^62,63 C. roseus compounds were found to directly bind to several proteins, including AKT, VEGF, EGF, eNOS, FGF, HIF-1α, IL-1β, IL-10, IR, IRS-1, MCP-1, MMP-9, NF-κB, PDGF, SDF-α, SOD, TGF-β1, and TF-α. IL-10, TGF-β1, EGF, eNOS, MMP-9, and Nrf2 with strong hydrogen bonds. Effective interactions were shown by compounds with binding energies, including staphyloxanthin, vinblastine, and vincristine. Compared to other compounds, vincristine showed the strongest binding, particularly with IL-10, and formed stable interactions through multiple hydrogen bonds, with key residues such as GLU, ARG, ASN, and TYR playing a crucial role. These interactions highlighted the compounds’ potential to modulate growth factors and inflammatory pathways, which makes them potential therapies for diabetic wound healing (Tables 4 & 5). In vivo studies further supported these findings, showing that the ethanol extract of C. roseus accelerated wound healing in diabetic rats by enhancing angiogenesis, epithelization, antimicrobial activity, and tensile strength. It also improved biochemical markers such as hexuronic acid, hydroxyproline, hexosamines, lysyl oxidase, and tissue protein levels, indicating its potential as a topical therapeutic.^10,27 MD simulations confirmed the stability of these interactions, with the MMP-9–STX complex maintaining RMSD values around ∼0.14–0.17 nm, while the Serine Protease–VBL complex displayed even greater stability (∼0.11-0.13 nm). RMSF analysis revealed that fluctuations were limited mainly to loop and terminal regions, while residues at the binding sites remained stable. These results suggest that C. roseus phytochemicals can effectively suppress proteolytic activity associated with diabetic wound pathogens. Collectively, these findings highlight their role in regulating the TNF-α/VEGF signalling axis to promote angiogenesis and wound repair. Overall, C. roseus exhibits significant therapeutic potential in diabetic wound healing via antioxidant, anti-inflammatory, and antidiabetic properties. Its effects are reinforced by enzyme inhibition, antimicrobial activity, and molecular-level interactions with wound-healing targets. Nevertheless, further studies needed to confirm the safety and efficacy in humans. Future research should prioritize well-designed trials to validate these findings and fully establish their therapeutic applications.

Limitation of the Study

Despite promising in vitro and in silico results, this study has certain limitations. Docking and short-term molecular dynamics simulations provide predictive insights but may not fully represent the complex diabetic wound microenvironment. The findings are limited to in vitro assays, with no in vivo wound-healing validation. Although vincristine and vinblastine showed strong target affinity, cytotoxicity and dermal safety evaluations need to be evaluate. Further in vivo, safety, and mechanistic studies are required to establish clinical relevance.

Conclusion

C. roseus shows remarkable potential in diabetic wound healing due to diverse phytochemical profile, including alkaloids, flavonoids, and tannins. The bioactive compounds, implicated in modulating the NF-κB and VEGF pathways. These metabolites exhibited strong antioxidant, anti-inflammatory, and antimicrobial properties, which collectively promote angiogenesis, reduce inflammation, and accelerate tissue regeneration. Molecular docking studies further supported its therapeutic role by demonstrating interactions with key wound-healing proteins, indicating its capacity to regulate oxidative stress, improve glucose metabolism, and enhance wound closure. Specifically, vincristine and vinblastine showed strong binding with microbial enzymes, suggesting inhibition of pathogen virulence factors and modulation of TNF-α and VEGF signalling. Together, these findings highlight C. roseus as a cost-effective natural candidate for diabetic wound management, though further investigations essential to validate its safety and therapeutic efficacy in humans.^64,65

Supplemental Material

sj-docx-1-npx-10.1177_1934578X261433097 - Supplemental material for Phytochemical and Biological Investigation of Catharanthus roseus: Diabetic Wound-Healing Approach

Supplemental material, sj-docx-1-npx-10.1177_1934578X261433097 for Phytochemical and Biological Investigation of Catharanthus roseus: Diabetic Wound-Healing Approach by Mohammad A. Alshuniaber, Magdi A. Osman and Rajapandiyan Krishnamoorthy in Natural Product Communications

Footnotes

Acknowledgments

The authors thank the Ongoing Research Funding Program, (ORF-2026-1226), King Saud University, Riyadh, Saudi Arabia, for funding this research.

ORCID iDs

Mohammad A. Alshuniaber

Rajapandiyan Krishnamoorthy

Ethics Approval and Informed Consent

This article does not contain any studies with human participants or animals performed by any of the authors.

Consent for Publication

Not applicable

Authors’ Contribution

MA; Project concept developed, Funding and supervising, MO; data validations, Original Manuscript writing and review, RK: Experimental, data validations, Original Manuscript writing and review,

Funding

The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: Ongoing Research Funding Program, (ORF-2026-1226), King Saud University, Riyadh, Saudi Arabia.

Declaration of Conflicting Interests

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Data Availability Statement

The datasets used during the current study available from the corresponding author on reasonable request

Supplemental Material

Supplemental material for this article is available online.

References

Kumar

Saha

Kumar

Sahana

Dubey

Prakash

. A review on diabetes mellitus: type1 & Type2. World J Pharm Pharm Sci. 2020;9(10):838-850.

Attah

Jacks

Jacob

Eduitem

John

. The effect of aloe vera (linn) on cutaneous wound healing and wound contraction rate in adult rabbits. Nova J Med Biol Sci. 2016;5(3):1-8.

Arise

Akapa

Adigun

Yekeen

Oguntibeju

. Normoglycaemic, normolipidemic and antioxidant effects of ethanolic extract of Acacia ataxacantha root in STZ-induced diabetic rats. Not Sci Biol 2016;8(2):9970. doi:10.15835/nsb.8.2.9770

Nair

Gowtham

Paridhavi

. Some promising medicinal plants with anti-diabetic wound healing activity: a review. Asian J Pharm Pharmacol. 2018;4(4):379-385. doi:10.31024/ajpp.2018.4.4.1

Okonkwo

DiPietro

. Diabetes and wound angiogenesis. Int J Mol Sci. 2017;18(7):1419. doi:10.3390/ijms18071419

Kloesch

Becker

Dietersdorfer

Kiener

Steiner

. Anti-inflammatory and apoptotic effects of the polyphenol curcumin on human fibroblast-like synoviocytes. Int Immunopharmacol. 2013;15(2):400-405. doi:10.1016/j.intimp.2013.01.003

Chereddy

Coco

Memvanga

, et al. Combined effect of PLGA and curcumin on wound healing activity. J Control Release. 2013;171(2):208-215. doi:10.1016/j.jconrel.2013.07.015

Peña

Martin

. Cellular and molecular mechanisms of skin wound healing. Nat Rev Mol Cell Biol. 2024;25(8):599-616. doi:10.1038/s41580-024-00715-1

Guo

Dipietro

. Factors affecting wound healing. J Dent Res. 2010;89(3):219-229. doi:10.1177/0022034509359125

10.

Nayak

. Influence of ethanol extract of Vinca rosea on wound healing in diabetic rat. Online J Biol Sci. 2006;2(6):51-55. doi:10.3844/ojbsci.2006.51.55

11.

Shaik-Dasthagirisaheb

Varvara

Murmura

, et al. Vascular endothelial growth factor (VEGF), mast cells and inflammation. Int J Immunopathol Pharmacol. 2013;26(2):327-335. doi:10.1177/039463201302600206

12.

Mangoni

McDermott

Zasloff

. Antimicrobial peptides and wound healing: biological and therapeutic considerations. Exp Dermatol. 2016;25(3):167-173. doi:10.1111/exd.12929

13.

Perez Gutierrez

Vargas

. Evaluation of the wound healing properties of Acalypha langiana in diabetic rats. Fitoterapia. 2006;77(4):286-289. doi:10.1016/j.fitote.2006.03.011

14.

Lin

Lan

Lin

. Expression of miR-217 and HIF-1α/VEGF pathway in patients with diabetic foot ulcer and its effect on angiogenesis of diabetic foot ulcer rats. J Endocrinol Invest. 2019;42(11):1307-1317. doi:10.1007/s40618-019-01053-2

15.

Patel

Vadgama

Baxi

Chandrabhanu, Tripathi

. Evaluation of hypolipidemic activity of leaf juice of Catharanthus roseus (linn.) G. Donn. In Guinea pigs. Acta Pol Pharm. 2011;68(6):927-935.

16.

Alxeandrova

Velcheva

Varadino

. Phytoproducts and cancer. Exp Patho Parasitol. 2000;4:15-25.

17.

van Der Heijden

Jacobs

Snoeijer

Hallard

Verpoorte

. The Catharanthus alkaloids: pharmacognosy and biotechnology. Curr Med Chem. 2004;11(5):607-628. doi:10.2174/0929867043455846

18.

Das

Sharangi

. Madagascar Periwinkle (Catharanthus roseus L.): diverse medicinal and therapeutic benefits to humankind. J Pharmacogn Phytochem. 2017;6(5):1695-1701.

19.

Nisar

Mamat

Hatim

Aslam

Syarhabil

. An updated review on Catharanthus roseus: phytochemical and pharmacological analysis. Indian Res. J. Pharm. Sci. 2016;3(2):631-653.

20.

Patil

Ghosh

. Antimicrobial activity of Catharanthus roseus–a detailed study. BJPT. 2010;1(1):40-44.

21.

Bhutkar

Bhise

. Comparative studies on antioxidant properties of Catharanthus rosea and Catharanthus alba. Int. J. Pharmtech Res. 2011;3(3):1551–1556.

22.

Kokate

Purohit

Gokhale

. Text book of pharmacognosy. Pune: Nirali Prakashan. 2003;8(66):1-624.

23.

Sain

Sharma

. International journal of pure & applied bioscience catharanthus roseus (an anti-cancerous drug yielding plant)—a review of potential therapeutic properties. Int. J. Pure App. Biosci. 2013;1(6):139-142.

24.

Singh

. Antidiabetic and wound healing activity of Catharanthus roseus L. in streptozotocin-induced diabetic mice. Am J Phytomed Clin Ther. 2014;2(6):686-692.

25.

Al-Shaqha

Khan

Salam

Azzi

Chaudhary

. Anti-diabetic potential of Catharanthus roseus Linn. and its effect on the glucose transport gene (GLUT-2 and GLUT-4) in streptozotocin induced diabetic wistar rats. BMC Complement Altern Med. 2015;15:379. doi:10.1186/s12906-015-0899-6

26.

Patel

Vadgama

Baxi

Chandrabhanu, Tripathi

. Evaluation of hypolipidemic activity of leaf juice of Catharanthus roseus (Linn.) G. Donn. In Guinea pigs. Acta Pol Pharm. 2011;68(6):927-935.

27.

Saravanan

Pichaivel

Venkatesan

Krishnaraju

Paulsamy

Kuppan

. Wound healing potential of Catharanthus Roseus on dead space wound in diabetic rats. World J. Pharm. Res.. 2021;9(8):92–96.

28.

Singh

Kumar

. Study of phytochemical analysis and antioxidant activity of Allium sativum of Bundelkhand region. Int. J. Pure App. Biosci. 2017;3(6):1451-1458.

29.

Audu

Mohammed

Kaita

. Phytochemical screening of the leaves of Lophira lanceolata (Ochanaceae). Life Sci J. 2007;4(4):75-79.

30.

Sheel

Nisha

Kumar

. Preliminary phytochemical screening of methanolic extract of Clerodendron infortunatum. IOSR-JAC. 2014;7(1):10-13. doi:10.9790/5736-07121013

31.

Kancherla

Dhakshinamoothi

Chitra

Komaram

. Preliminary analysis of phytoconstituents and evaluation of anthelminthic property of Cayratia auriculata (in vitro). Maedica (Bucur). 2019;14(4):350-356.

32.

Sakthi

Geetha

Saranraj

. Pharmacological screening of Datura metel and Acalypha indica for its antifungal activity against pathogenic fungi. IJPHC. 2011;2(1):15-30.

33.

Silva

Abeysundara

Aponso

. Extraction methods, qualitative and quantitative techniques for screening of phytochemicals from plants. Am J Essent Oil Nat Prod. 2017;5(2):29-32.

34.

Sawant

Godghate

. Qualitative phytochemical screening of rhizomes of Curcuma longa Linn. Int J Sci Environ Technol. 2013;2(4):634-641.

35.

Basumatary

. Preliminary phytochemical screening of some compounds from plant stem bark extracts of Tabernaemontana divaricata Linn. used by Bodo community at Kokrajhar District, Assam, India. Arch. Appl. Sci. Res. 2016;8(8):47-52.

36.

Shaikh

Patil

. Qualitative tests for preliminary phytochemical screening: an overview. Int J Chem Stud. 2020;8(2):603-608. doi:10.22271/chemi.2020.v8.i2i.8834

37.

Okhale

Oladosu

Aboh

Imoisi

Gana

. In-vitro evaluation of Eucalyptus citriodora leaf essential oil and extracts on selected pathogens implicated in respiratory tract infections. Int. J. Pharmacogn. 2022;9(12):195-201.

38.

Patel Rajesh

Patel Natvar

. In vitro antioxidant activity of coumarin compounds by DPPH, super oxide and nitric oxide free radical scavenging methods. JAPER. 2011;1(1):52-68.

39.

Gunathilake

KDPP

Ranaweera

KKDS

Rupasinghe

HPV

. In vitro anti-inflammatory properties of selected green leafy vegetables. BIOMID. 2018;6(4):107.

40.

Mechchate

Es-Safi

Louba

, et al. In vitro alpha-amylase and alpha-glucosidase inhibitory activity and in vivo antidiabetic activity of Withania frutescens L. Foliar Extract Molecules. 2021;26(2):293.

41.

Young

McNaught

. The physiology of wound healing. Surgery (Oxf). 2011;29(10):475-479. doi:10.1016/j.mpsur.2011.06.011

42.

Ali

Rathore

Sharma

. Modern review on diabetic wound healing potential of some medicinal herbs. Int J Pharm Sci. 2024;Vol 2(Issue 2):330-338.

43.

Duflos

Kruczynski

Barret

. Novel aspects of natural and modified vinca alkaloids. Curr Med Chem Anticancer Agents. 2002;2(1):55-70. doi:10.2174/1568011023354452

44.

Bennouna

Delord

Campone

Nguyen

. Vinflunine: a new microtubule inhibitor agent. Clin Cancer Res. 2008;14(6):1625-1632. doi:10.1158/1078-0432.CCR-07-2219

45.

Mishra

Verma

. A brief study on Catharanthus roseus: a review. Int J Res Pharm Pharm Sci. 2017;2(2):20-23.

46.

Kumari

Gupta

. Phytopotential of Catharanthus roseus L.(G.) Don. var.“Rosea” and “Alba” against various pathogenic microbes in vitro. Int J Res Pure Appl Microbiol. 2013;3(3):77-82.

47.

Veerichetty

Saravanabavan

Pradeep

. Identification of bioactive compounds by GC-MS of nelumbo Nucifera leaf extract and virtual screening of EGFR/VEGFR2 dual inhibitors. Biosci Biotechnol Res Asia. 2024;21(3):821. doi:10.13005/bbra/3267

48.

Parveen

Subedi

Kim

, et al. Phytochemicals targeting VEGF and VEGF-related multifactors as anticancer therapy. J Clin Med. 2019;8(3):350. doi:10.3390/jcm8030350

49.

Fakhri

Abbaszadeh

Jorjani

Pourgholami

. The effects of anticancer medicinal herbs on vascular endothelial growth factor based on pharmacological aspects: a review study. Nutr Cancer. 2021;73(1):1-15. doi:10.1080/01635581.2019.1673451

50.

Aziz

Saha

Sultana

Khan

Nada

Afroze

. Comparative studies of volatile components of the essential oil of leaves and flowers of Catharanthus roseus growing in Bangladesh by GC-MS analysis. Indian J Pharm Biol Res. 2015;3(1):6. doi:10.30750/ijpbr.3.1.2

51.

Mohan

Anand

Priyadharshini

Balamurugan

. GC-MS analysis of phytochemicals and hypoglycemic effect of Catharanthus roseus in alloxan-induced diabetic rats. Int J Pharm Sci Rev Res. 2015;31(1):123-128.

52.

Alkreathy

Ahmad

. Catharanthus roseus combined with ursolic acid attenuates streptozotocin-induced diabetes through insulin secretion and glycogen storage. Oxid Med Cell Longev. 2020;2020:8565760. doi:10.1155/2020/8565760

53.

Rasineni

Bellamkonda

Singareddy

Desireddy

. Antihyperglycemic activity of Catharanthus roseus leaf powder in streptozotocin-induced diabetic rats. Pharmacogn. Res. 2010;2(3):195-201. doi:10.4103/0974-8490.65523

54.

Chattopadhyay

Sarkar

Ganguly

Banerjee

Basu

. Hypoglycemic and antihyperglycemic effect of leaves of Vinca rosea linn. Indian J Physiol Pharmacol. 1991;35(3):145-151.

55.

Singh

Vats

Suri

, et al. Effect of an antidiabetic extract of Catharanthus roseus on enzymic activities in streptozotocin induced diabetic rats. J Ethnopharmacol. 2001;76(3):269-277. doi:10.1016/S0378-8741(01)00254-9

56.

Dash

Sultana

. Antibacterial activities of methanol and acetone extracts of fenugreek (Trigonella foenum) and coriander (Coriandrum sativum). LSMR. 2011;27(1):1-8.

57.

Hemanth

Vimal

. Assessment of anti-oxidative, anti-inflammatory, and anti-cancer activity of magnesium oxide doped chitosan/polyvinyl alcohol with Catharanthus roseus: an in vitro study. Cureus. 2024;16(9):e70103.

58.

Kabesh

Senthilkumar

Ragunathan

Kumar

. Phytochemical analysis of Catharanthus roseus plant extract and its antimicrobial activity. Int J Pure App Biosci. 2015;3(2):162-172.

59.

Kaur

Mondal

. Study of total phenolic and flavonoid content, antioxidant activity and antimicrobial properties of medicinal plants. J Microbiol Exp. 2014;1(1):00005.

60.

Chattopadhyay

. A comparative evaluation of some blood sugar lowering agents of plant origin. J Ethnopharmacol. 1999;67(3):367-372. doi:10.1016/S0378-8741(99)00095-1

61.

Sultana

Nibir

Ahmed

. Biosensing and anti-inflammatory effects of silver, copper and iron nanoparticles from the leaf extract of Catharanthus roseus. BJBAS. 2023;12(1):26.

62.

Deepika

Santhy

. In vitro antioxidant and antidiabetic activity of silver nanoparticles synthesized using Catharanthus roseus leaves. RJPT. 2022;15(3):989-997.

63.

Mariappan

pk Jabir

Babu

Krishnan

. Unveiling the Promise of Bioactive Alkaloid Compound from Catharanthus Roseus: An In-vitro Computational Exploration of their Molecular Docking against a Target Protein for Type-2 Diabetes. J Clin Diagn Res. 2024;18(8):1–4.

64.

Kobayashi

Koguchi

Takahashi

Kanzaki

Kawazu

. ST-1, a novel radical scavenger from fungus F-124. Biosci. Biotechnol. Biochem. 1993;57(6):1034-1036. doi:10.1271/bbb.57.1034

65.

Selvaraj

. Identification of new antidiabetic agents targeting GLUT4 protein using in silico analysis. IJGP. 2018;12(04). doi:10.22377/ijgp.v12i04.2269

Supplementary Material

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Phytochemical and Biological Investigation of Catharanthus roseus : Diabetic Wound-Healing Approach

Abstract

Objectives

Methods

Results

Conclusion

Keywords

Introduction

Materials and Methods

Chemicals and Reagents

Extract Preparation

Phytochemical Screening

GC-MS Analysis

Antioxidant Activity-DPPH Radical Scavenging Assay

Anti-inflammatory Activity by Protein Denaturation Assay

Anti-diabetic Activity: α-Amylase & α-Glucosidase Inhibitory Activity

Antimicrobial Activity: Determination of Minimum Inhibitory Concentration (MIC)

Molecular Docking

Molecular Dynamics

Statistical Analysis

Results

Phytochemical Analysis

GC-MS Analysis of a Methanolic Extract of Catharanthus Roseus

Effects of Antioxidant DPPH Activity of Extract

Effects of Anti-Inflammatory Activity of Extract

Effects of α-Amylase Inhibition Activity of Extract

Effects of α-Glucosidase Inhibition Activity of Extract

Minimum Inhibitory Concentration

In-silico Analysis - Docking Studies with Microbial Proteins

Docking Studies with Wound Healing Proteins

Molecular Dynamics

Discussion

Limitation of the Study

Conclusion

Supplemental Material

sj-docx-1-npx-10.1177_1934578X261433097 - Supplemental material for Phytochemical and Biological Investigation of Catharanthus roseus: Diabetic Wound-Healing Approach

Footnotes

Acknowledgments

ORCID iDs

Ethics Approval and Informed Consent

Consent for Publication

Authors’ Contribution

Funding

Declaration of Conflicting Interests

Data Availability Statement

Supplemental Material

References

Supplementary Material