Sage Journals: Discover world-class research

Abstract

Wogonoside is a flavonoid, the main effective constituent of Scutellaria baicalensis, belongs to the Lamiaceae family, with various functions, including detoxification, anti-inflammation and nourishing gallbladder, lowering blood pressure, diuresis and anti-allergic reactions. This study was designed to evaluate the positive role of Wogonoside (WS) on streptozotocin (STZ)-induced diabetes mellitus in rats. Rats were randomized into six groups: Normal control, STZ (60 mg/kg) induced diabetic rats, STZ + WS (100 mg/kg), STZ + WS (200 mg/kg), STZ + Glibenclamide (600 µg/kg) and WS alone (200 mg/kg). After 45 days, the plasma glucose, glycated hemoglobin (HbAC1), insulin level, carbohydrate metabolizing enzyme (Glucokinase, Glucose 6-Phosphatase, Fructose 1,6 -bisphosphatase and Glucose 6-phosphate dehydrogenase), biochemical parameters such as Aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, uric acid, creatinine, total cholesterol, triglycerides, phospholipids, Free fatty acids, Low-density Lipoprotein-C and High-density Lipoprotein-C and antioxidant enzymes status were measured. A pathological condition of the liver, kidney and pancreas was examined by hematoxylin-eosin (H&E) staining. After 45 days of treatment, the results showed that WS had significant antihyperglycemic and total cholesterol-lowering effects and improved serum high-density lipoprotein (HDL) cholesterol levels. WS also enhanced glucose tolerance and the histological state of the liver in diabetic rats and exhibited intriguing antioxidant activities.

Keywords

Wogonoside diabetes mellitus glucose insulin

Introduction

As per World Health Organization (WHO), around 422 million people are getting affected by Diabetes mellitus (DM), which includes 1.6 million deaths in each year from low and middle-income countries. Followed by WHO, the International Diabetes Federation (IDF), and its survey projected that more than 552 million people worldwide would be affected by DM within a decade.¹ Hence it is not only a noticeable disease from carbohydrate metabolism. It is also a reason to develop many associated causes like cardiac arrest, hypertension, kidney damage, and retinopathies due to defect in poor insulin secretion or mutation in Glucose transporter (GLUT) receptor of insulin hormone in beta cells of Langerhans of the pancreas.² The hepatic organ of the liver is a major organ that controls the intermediary metabolism of our body and supplies energy in the form of ATP. The oxidation of glucose synthesis ATP as glycolysis, the amphibolic role of Tricarboxylic acid cycle (TCA) as glycogenolysis, and the gluconeogenesis process are attained.³ For all these processes liver is the site and controls the metabolism. But, in the case of diabetes patients, the energy supply by the liver is get affected, and by the way, it interrupts liver functions by influencing carbohydrate metabolic enzymes.⁴ Gluco-6-phosphatase is the major rate-limiting enzyme in gluconeogenesis and glycogen metabolism were synthesis glucose in need of the body. In diabetes, the synthesis of glucose by glucose-6-phosphatase was two folded which was reverse and became worst as a severe hyperglycemic state and failed to activate glucokinase enzyme.⁵

Research reveals that some drugs are used as hypoglycemic agents to reduce hyperglycemic condition by maintaining blood's glycemic index. But those allopathic hypoglycemic drugs show contrary effects like lactic acidosis, fatty liver, and digestion problem. Also, those adverse effects were caused because of poor pharmacodynamics and pharmacokinetics management.⁶ A substantial prevalence is needed to treat, or maintenance is enough to control this DM. As per research, DM disease cannot be treated diseases, meantime only by precaution methods and control management by medications are available. The recovery of insulin secretion has not yet been achieved, and the exact therapy/medication is not yet identified. However, diet management, continuous physical exercise, and yoga treatment are used to avoid hyperglycemia and related issues.⁷ What follows is not enough to overcome hyperglycemic conditions due to time management, lifestyle, and work pressure. Although, our society needs a drug that is without or less adverse effects to overcome the International Diabetes Federation (IDF) survey was mentioned above.

Out present research is focused on developing an antidiabetic drug free of side effects, easy availability, and less cost. Plants are readily available with less cost, and under-developing countries can also access or discover a drug for diabetes based on previously reported results. Plants and each part of the plant have some medicinal properties and are used as antibacterial, antifungal, anticancer, antimetabolites agents.^8-10 Recently, developing an anti-diabetic drug is challengeable with fewer side effects and its approval by the drug control board. However, the existing anti-diabetic drugs meet the controlling hyperglycemic condition of diabetes but fail to maintain associated secondary metabolic disorders like hypertension, liver problems, and kidney damage. Therefore, finding natural drugs from the medicinal plant with better anti-diabetic function is the significant scope against diabetes mellitus.

Materials and Methods

Plant Material and Chemicals

Wogonoside (51059-44-0) obtained from Sigma Aldrich pvt Ltd, China. Streptomycin, Glibenclamide were purchased from Sigma (St. Loius, MO, USA). Rat insulin ELISA kit was purchased from EMD Millipore Corporation (USA). Superoxide Dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) and malondialdehyde (MDA) kit were all procured from Nanjing Jiancheng Bioengineering Institute (China). All the other compounds utilized in this experiment were analytical grade.

Ethics

The present study was approved by the Institutional Animal Ethical Committee (IAEC) of Hubei University of Medicine, P.R. China with the approved ethical accession number 2021887789. As per IAEC approval, the both male and female Wistar rats (7-9 weeks old; with an average 200 g of weight) was attained from Wuhan Institute of Biological Products Co. (Wuhan, China). The rats were kept at normal temperature (25 °C) and maintained as 12 h light/12 h dark cycle with a normal diet with water supply through libitum.

Induction of Diabetes and Experimental Design of Study Group's

For diabetic conditions, the rats were administered streptozotocin (STZ) (diluted in M sodium citrate buffer with pH of 4.5) through a once in a day of five-shot of intraperitoneal (IP) injection as per 50 mg/kg body weight.¹² Also, to restrict the drug-induced hypoglycemic condition in STZ-induced rats were orally administered with 20% of glucose in solution. After, the STZ-induced rats were allowed to maintain hyperglycemic conditions for a 45 days. Then, the rats were confirmed their diabetic condition of exhibited plasma glucose level is above 250 mg/dL.¹³ The diabetic condition of rats was confirmed and carried for further experiments.

The Rats were categorized into six groups, with each group containing six rats:

Group 1: Normal control (With normal chow feed)

Group 2: STZ (60 mg/kg) induced DN rats (Negative control)

Group 3: STZ + Wogonoside (100 mg/kg)

Group 4: STZ + Wogonoside (200 mg/kg).

Group 5: STZ + Glibenclamide (600 µg/kg)

Group 6: Wogonoside alone (200 mg/kg)

Wogonoside was dissolved in 5% Dimethyl sulfoxide (DMSO). The known drug Glibenclamide was dissolved in water and orally administered with the help of an intra-gastric tube for 45 days to all the study groups and maintained. At the end of the 45 days of the experimental study, the final body weight was measured and was compared with the initial body weight. Also, the animals were prepared for sacrifices, and before that, the animals were kept under overnight fasting; ketamine (12 mg/kg) was injected through the IM route to produce anesthesia in rats. The blood samples were collected directly from heart puncture during the sacrifice in the anti-coagulant coated (sodium citrate and sodium fluoride in the 3:1 ratio) tube. Blood samples processed and used to assess the insulin, plasma glucose, and other biochemical parameters quantification. After sacrifice (cervical dislocation), immediately the tissues were collected then washed twice with ice-cold hyposaline (pH7.4). After wash, the tissues were homogenized under ice-cold conditions followed by centrifugation. Then, the upper layer of supernatant was collected for various liver, kidney markers and biochemical marker analysis.

Biochemical Parameters

Glucose and Glycosylated hemoglobin are essential biochemical, and carbohydrate metabolic defect parameters estimated by the kit method (AccuChek, Basel, Switzerland and R&D, Minneapolis, MN, USA). The plasma insulin level was quantified by the enzymatic ELIZA kit method. After insulin, we need to check the carbohydrate metabolic rate-limiting enzymes changes to compare whether the drug influences the metabolism to improve/decay the cells.¹⁴ So, the following rate-limiting enzymes of (Hexokinase, Glucose-6-phosphatases, Fructose, 1.6 bi-phosphatases) carbohydrate metabolism were assessed by an enzymatic kit method as per manufacturer instructions. Followed by enzymes, diabetic kidney markers such as urea, uric acid, and creatinine were assessed using standard Diacetyl monoxime and Jeff's method. Meantime, plasma glucose, HbA1c, Uric acid, and creatinine of primary diabetic parameters were estimated and compared with the fasting state of experimental rats and compared after the study period and to validate for further experiments.

Lipids Estimation

The total cholesterol estimation was done by standard Zak's method, and triglycerides (TG) were measured using an enzymatic kit purchased from Sigma Aldrich. Like TG, total phospholipids (PL) and free fatty acids (FFA) were also estimated by an enzymatic kit method. Lipids carrying lipoproteins such as low-density lipoprotein (LDL) and high-density lipoprotein (HDL) was assessed by non-enzymatic kit methods as per Sigma Aldrich manufacturer.

Liver Marker Analysis

Liver markers such as serum alanine transaminases (ALT) and serum aspartate transaminases (AST) were assessed by standard methods of enzymatic UV-kinetic and Reitman-Frankel method.

Antioxidant's Analysis

Standard Marklund and Marklund methods assessed enzymatic antioxidants such as superoxide dismutase (SOD). Catalase was assessed by Aebi et al and the enzymatic Paglia and Valentine method estimated glutathione peroxidase (GPx). Meanwhile, non-enzymatic antioxidants such as Vit- E (alpha-tocopherol) were carried out by standard HPLC procedure as per Aitken et al Finally, reduced glutathione was processed by kit and finally estimated by colourimetric analysis.

Histopathology

Wogonoside were treated, and histopathological studies assessed its toxicity on pancreas, kidney, liver, and morphological changes after 45 days of treatment were performed by investigating its cross-tissue sections of organs were embedded with paraffin seal was stored and used for further examination. In brief, the collected organs from the immediate sacrifice were stored in 10% neutral formalin buffer embedded with paraffin seal tissues, cut into 5 micrometer thick sections, and stained with hematoxylin and eosin (H and E). Once the tissue sections were fit with H and E stain was viewed under a light microscope (original magnification is ×40)

Statistics

Statistical analysis was performed using student t-test was used to compare between different samples. P < .05 was considered as significant. Results were expressed as mean ± standard deviation (SD). Tukeys post-hoc statistical power analysis was performed and values obtained were >0.8 which indicate adequate sample size.

Results

Table 1 explains the effect of WS on glucose and insulin levels in normal and STZ induced diabetic rats. Totally six groups were selected and experimented with. There is an increase in plasma glucose and insulin level with noticeable significant was found in the group 3, 4 and 5 were found than negative control (STZ group 2). At the same time, WS alone (200 mg/kg) treated group rats were found to increase in plasma glucose and insulin level on the first day of the experiment (Group 6). At the end of the study, they maintained the plasma glucose and insulin as equal to the control group. Hence, the dosage of WS alone (200 mg/kg) was selected for further biochemical and marker studies. Glycosylated haemoglobin of HbA1c levels in control and treated rats were depicted in Table 1. Reduced HbA1c level was noted with P < .05 significant in WS alone (200 mg/kg) dosage treated diabetic rats than group 4 and 5. Also, the found HbA1c levels were almost like normal control rats of experimental studies.

Table 1.

Effect of WS on HbAC1, Insulin and Glucose Levels in the Normal and STZ Induced Diabetic Rats.

Groups	HbAC1 (%)	Insulin (µU/mL)	Plasma glucose(mg/dL)
Groups	HbAC1 (%)	Insulin (µU/mL)	Initial day	End day
Control	7.23 ± 1.36^a	15.05 ± 1.05^a	87.31 ± 7.22^a	85.24 ± 6.81^a
STZ (60 mg/kg)	15.93 ± 0.92^e	6.91 ± 1.20^e	255.20 ± 15.64^b	301.14 ± 22.84^e
STZ + WS (100 mg/kg)	13.89 ± 1.91^d	8.02 ± 0.74^d	250.68 ± 18.74^b	190.57 ± 17.54^d
STZ + WS (200 mg/kg)	11.36 ± 1.78^c	10.09 ± 0.91^c	258.57 ± 19.54^b	124.85 ± 10.45^c
STZ + Glibenclamide (600 µg/kg)	8.21 ± 0.25^b	13.01 ± 0.86^b	252.38 ± 12.56^b	110.32 ± 5.43^b
WS alone (200 mg/kg)	7.91 ± 1.52^a	15.64 ± 0.52^a	257.52 ± 13.94^b	89.91 ± 3.60^a

Data are mean ± SEM values, n = 6. Data were analyzed by One way ANOVA followed by Turkey Kramer Multiple comparisons test. The values superscript letters mean significant differences at P < .05. WS: Wogonoside, ^aWhen compared with normal control group, ^b when compared with inducer group, ^c when compared with Standard group, ^d when compared with WS Alone and ^e when compared with STZ + WS (100 mg/kg).

Table 2 explains that significant regulating enzymes of carbohydrate metabolism were represented here mean ± standard deviation. The enzymes of glucokinase, glucose-6-phosphatase, fructose 1,6 bisphosphatase, and glucose 6 phosphate dehydrogenase were found with less in diabetic rats treated with STZ (Group 2) induced Glibenclamide (Group 4), and WS (Group 5) than control and WS alone treated experimental studies. Hence, when compared to STZ with WS, WS alone treated was given better significance (P < .05) and almost near to normal control rats. Again, these results prove that WS alone worked better on diabetic rats.

Table 2.

Effect of Wogonoside on the Activity of Carbohydrate Metabolic Enzymes in the Liver of Control and STZ Induced Diabetic Rats.

Groups	Glucokinase (U*/mg protein)	Glucose 6-phosphatase (Unit#/mg protein)	Fructose 1,6 -bisphosphatase (Unit$/mg protein)	Glucose 6-phosphate dehydrogenase (Unit@/ mIU /mg protein)
Control	0.350 ± 0.03^a	0.230 ± 0.02^a	0.425 ± 0.01^a	4.70 ± 0.35^a
STZ (60 mg/kg)	0.145 ± 0.02^d	0.450 ± 0.04^d	0.730 ± 0.05^d	2.10 ± 0.12^d
STZ + WS (200 mg/kg)	0.234 ± 0.02^c	0.360 ± 0.03^c	0.554 ± 0.04^c	3.06 ± 0.21^b
STZ + Glibenclamide (600 µg/kg)	0.284 ± 0.02^b	0.277 ± 0.02^b	0.501 ± 0.01^b	2.74 ± 0.31^c
WS alone (200 mg/kg)	0.347 ± 0.01^a	0.229 ± 0.05^a	0.446 ± 0.02^a	4.83 ± 0.26^a

Data are mean ± SEM values, n = 6. Data were analyzed by One way ANOVA followed by Tukey Kramer Multiple comparisons test. The values superscript letters mean significant differences at P < .05. WS: Wogonoside, ^aWhen compared with normal control group,^b when compared with inducer group, ^c when compared with Standard group, ^d when compared with WS Alone and ^e when compared with STZ + WS (100 mg/kg).

The kidney and liver function conditions were tested by measuring urea, uric acid, creatinine, AST, and ALT, as depicted in Table 3. The increased activity of AST and ALT was obtained in diabetic rats than control. At the same WS alone (Group 5) treated experimental rats showed uncontrolled in an initial day. At the end of experimentation was become almost normal like control groups with a P < .05 significant level. On the other hand, urea, uric acid, and creatinine were highly excreted in the serum of diabetic rats (Group 2). However, STZ + WS alone treated rats was significant (P < .05) with maintained serum markers was also depicted in table 3 (Group 5).

Table 3.

Effect of Wogonoside on serum Hepatic and Plasma Renal Markers in the serum of Control and STZ Induced Diabetic Rats.

Groups	Liver markers		Kidney markers
Groups	ALT (IU/L)	AST (IU/L)	Urea (mg/dL)	Uric acid (mg/dL)	Creatinine (mg/dL)
Control	30.21 ± 2.81^a	80.41 ± 7.89^a	26.54 ± 1.54^a	1.44 ± 0.21^a	1.01 ± 0.19^a
STZ (60 mg/kg)	65.24 ± 4.51^d	120.15 ± 09.54^d	39.46 ± 2.70^d	2.06 ± 0.12^d	2.62 ± 1.13^d
STZ + WS (200 mg/kg)	48.57 ± 2.89^c	97.75 ± 7.96^c	31.59 ± 2.51^c	1.35 ± 0.25^c	1.94 ± 0.23^c
STZ + Glibenclamide (600 µg/kg)	36.95 ± 1.54^b	91.48 ± 4.37^b	28.43 ± 1.06^b	1.26 ± 0.04^b	1.24 ± 0.02^b
WS alone (200 mg/kg)	32.41 ± 1.28^a	82.71 ± 2.63^a	25.89 ± 0.94^a	1.47 ± 0.19^a	1.04 ± 0.11^a

Table 4 was listed with important lipid parameters of lipid metabolism, which was significant when compared to diabetes patients. There are increased levels of serum total cholesterol, triglycerides, FFA, phospholipids, LDL, and HDL was found in diabetic rats (Group 2) than in control. But, in a group of STZ + WS (Group 3), its lipid parameters were brought back to normal state after WS alone treated experimental groups was significant with P < .05 of DMRT.

Table 4.

Effect of Wogonoside on Lipid Profile in the Plasma of Normal and STZ Induced Diabetic Rats.

Groups	Total Cholesterol (mg/dl)	Triglycerides (mg/dl)	Phospholipids (mg/dl)	Free fatty acids (mg/dl)	LDL-C (mg/dl)	HDL-C (mg/dl)
Control	78.21 ± 4.51^a	61.54 ± 5.21^a	87.19 ± 5.64^a	59.21 ± 5.64^a	19.18 ± 1.53^a	37.57 ± 2.54^a
STZ (60 mg/kg)	147.72 ± 10.54^d	164.41 ± 12.54^d	146.24 ± 10.99^d	109.60 ± 7.64^d	51.82 ± 4.17^d	25.01 ± 1.08^d
STZ + WS (200 mg/kg)	117.45 ± 8.67^c	135.24 ± 5.81^c	108.96 ± 8.45^c	95.04 ± 3.89^c	42.05 ± 2.64^c	27.84 ± 1.94^c
STZ + Glibenclamide (600 µg/kg)	90.14 ± 8.67^b	86.54 ± 5.81^b	91.08 ± 8.45^b	65.87 ± 3.89^b	30.64 ± 2.64^b	32.17 ± 1.94^b
WS alone (200 mg/kg)	80.96 ± 6.57^a	64.92 ± 5.48^a	90.53 ± 7.26^a	62.27 ± 4.86^a	20.75 ± 1.42^a	38.42 ± 2.05^a

Table 5 explains that significant regulating enzymes of carbohydrate metabolism were represented here mean ± standard deviation. The enzymatic antioxidants such as SOD, CAT, GPx and non-enzymatic antioxidants (Vitamin E and Reduced Glutathione) levels were significantly altered in STZ treated diabetic rats. Hence, when compared to STZ (Group 2) with WS(Group 3), WS alone (Group 5) treated was significantly improved the antioxidants status and almost near to normal control rats. Again, these results prove that WS alone worked better antioxidants profile in diabetic rats.

Table 5.

The Activities of Antioxidants Enzyme status in Plasma of Control and Experimental Animals in Each Group.

Groups	SOD (U*/ml)	CAT U^# /ml	GPx (U^♣/I)	Vitamin ‘E’ [mg/dl]	Reduced Glutathione (mg / dl)
Control	3.93 ± 0.27^a	0.65 ± 0.04^a	142.36 ± 10.57^a	1.32 ± 0.11^a	25.17 ± 1.81^a
STZ (60 mg/kg)	2.38 ± 0.14^c	0.47 ± 0.03^d	112.73 ± 15.72^d	0.97 ± 0.09^b	17.64 ± 2.04^b
STZ + WS (200 mg/kg)	2.86 ± 0.60^d	0.54 ± 0.04^c	123.46 ± 13.61^c	1.27 ± 0.14^a	20.38 ± 1.61^a
STZ + Glibenclamide (600 µg/kg)	3.05 ± 0.31^b	0.60 ± 0.02^b	138.90 ± 14.52^b	1.21 ± 0.10^c	23.55 ± 1.07^c
WS alone (200 mg/kg)	3.90 ± 0.42^a	0.67 ± 0.06^a	143.72 ± 10.93^a	1.30 ± 0.21^a	25.03 ± 1.12^a

Effect of WS treated diabetic rats, and its histological profile of pancreas was stained by H&E staining was depicted in Figure 1. Comparing the histological profile of all the experimental groups, STZ + WS treated animals shows that pancreatic cells overcome diabetes (improved beta cells and its cross-sectional area) like normal cells than STZ induced diabetic rats.

Figure 1.

The Photomicrograph of Hematoxylin-Eosin Staining of Liver Histopathology of Diabetic Rats. Normal Rats Showing Normal Hepatocytes and SZT Induced Diabetic Rats Exhibiting Fatty Change and Inflammatory Cell Infiltrate. Diabetic with WS (200 mg/kg) Treated Presenting Normal Hepatocytes and Glibenclamide Treatment Rats Showing Dilated Central Vein When Compared with Diabetic Rats.

Discussion

In recent years the development of diabetes worldwide has an uncontrollable. Also, middle-aged people are more prone to this disorder due to their trendy and modern lifestyle than other factors (genetic inheritance).^15,16 Meantime, the treatment/to maintain plasma glucose level under control for those numerous methods followed by those who affected people was advised by clinicians.¹⁷ But, due to short-being treatment or the side effects of those prescriptions, no proper medication/treatments were not yet identified. Hence, the world is still struggling with that DM. However, the research reveals that poor management in maintaining diabetes in people was challenging, especially in the near 50's ages. Although, people nowadays are fonder of natural medication/organic foods than allopathic to avoid side effects.¹⁸ Hence, the present study aims to come with the anti-diabetic management from the Wogonoside was proven by animal model. At first, the plant is chosen for the study was present with enough antioxidants (Terpenoids, triterpenoids, DPPH free radical scavenging activity, total antioxidant capacity) like an existing medicinal plant. It was compared with standard medications already followed in an animal trial as per protocol.

In type I diabetes, critical regulatory enzymes carbohydrate metabolism plays a significant role and cause much inborn error or autoimmune diseases. Also, in type II the deficiency of these enzymes causes uncontrolled diabetes and creates many associated diseases like fatty liver and retinopathy.¹⁹ Hence, the present study determined the effect of key regulatory enzymes and its efficacy by WS-treated was improved than diabetic induced experimental rats. As per research protocol, STZ is used to induce diabetes in the experimental animal. 40 mg/kg of one low dose of STZ was taken and injected intraperitoneally was enough to damage the beta cells of the pancreas then reduce the insulin secretion, which was confirmed with plasma glucose levels in type II diabetes.²⁰ Both type 1 and 2 diabetes are due to cause by the absence/insufficient insulin by the pancreas was reflected in blood plasma glucose as the hyperglycemic condition with altered biochemical, live, and kidney markers.^21,22 The present study and experimental rats induced with diabetes was shown with high plasma glucose and low insulin level. But in WS -treated rats, its plasma glucose was reduced than an earlier diabetic condition after the treatment. It was positively significant with increased plasma insulin levels of group 6 which denotes pancreatic beta secretion brought back by our medicinal plant. In detail arresting the movement of ATPase/K + channels in the pancreas which will affect the beta cells, degeneration was done by our like control drug glibenclamide function. Also, triterpenoids are significant antioxidants; anticancer activity has proved an inhibitor that inhibits sensitive ATPase/K + channels while regulating plasma glucose.²³ Hence, our Wogonoside is rich in flavonoids and worked on STZ-induced experimental rats like control.

Numerous in vivo and human clinical trials state that the middle-aged people are instructed to maintain their insulin level is normal, then only our body can be capable of sustaining homeostatic state as well as the amphibole role TCA cycle for a continuous supply of glucose by both liver and muscle gluconeogenesis and glycogenolysis.^24,25 Also, few reports state that it was due to a decrease in carbohydrate metabolic regulatory enzymes, and its negative association with tissue proteins might influence the person's body weight. However, in some cases, the insulin less secreting people will become obese, and in insulin absent people become lean was reported.^26-28 Hence, insulin influences the body weight (BMI) of diabetes and its associated disorder. Some of the preceding claims that lack ATP supply to tissue due to poor ATP production by carbohydrate metabolism will be a reason for poor structural protein and its synthesis.²⁹ Some of the preceding claims that lack ATP supply to tissue due to poor ATP production by carbohydrate metabolism will be a reason for poor structural protein and its synthesis. However, it may also be a reason for losing bodyweight due to low insulin versus negatively proportional /failure to oxidize glucose molecule behind the poor ATP synthesis.^30,31 Although, the present study and its results show that the STZ induced diabetic rats and are with after the oral administration of WS and was compared with the control drug. Meanwhile, the obtained results agreed with previous research and its management.^32-34 Moreover, the continuation insulin hormone controls protein metabolism by stimulating protein synthesis and delays protein catabolism. But lower insulin production reflects reduced structural protein synthesis with higher protein degradation, which was reversed and led to low hemoglobin – a blood protein synthesis.³⁴ As a result, unconditional glycation, which includes hemoglobin, albumin, LDL, fibronectin, and collagen, was directly related to unconditional diabetes. Murray et al reveal that low insulin levels damage the red blood cell protein called HbA1c of glycosylated hemoglobin is Hb diffuse to fuse with glucose and oxygen transportation carried function was damaged which was reflected by high HbA1c, ie 16% or >6.1/100 mg of blood than average level. At the same time, the life span of RBC and its connected HbA1C is enough to measure the patient's glycemic status. Therefore, HbA1c is the final validating marker test that will declare the patient belongs to diabetes or non-diabetes concerning management.³⁵ From the present study group, 6 of WS rated STZ – induced diabetic rats and its HbA1c level becomes normal like control than other experimental groups.

Glucokinase (GK) it's an essential irreversible rate-limiting enzyme in glycolysis and glycogen metabolism. GK's absence or impaired function and its results in diminished glucose oxidation by glycolysis lead to hyperglycemia with reduced ATP synthesis by carbohydrate metabolism.³⁴ Hence, this will make a way to non-enzymatic glycation of glucose as glucose -6-phosphate, which was catalyzed by phosphoglucoisomerase as an alternative pathway of glyoxalate cycle that synthesizes low ATP direct glycolysis. The release of GK was more minor in diabetic rats than oral administered WS group 6 rats.

The liver is a significant organ and has a significant function in maintaining homeostasis of blood glucose levels through carbohydrate metabolism. Also, all the major metabolisms took place in the liver than other organs.³⁶ During low glucose levels, glycogen metabolism and gluconeogenesis balance the supply of glucose. However, in diabetes conditions, poor insulin enhances the glucose -6- phosphatases (G6Pase) and fructose 1.6, bi phosphatases (F1, 6BPase). G6Pases is a critical glucogenic enzyme present in ER of integral protein, which acts as a dephosphorylating reaction of converting glucose 6- phosphate to glucose. But in the case of diabetes without oxidizing glucose molecules, due to poor recognition of insulin, G6Pase start to synthesis more glucose in diabetes than control.³⁷ Like G6Pase, F1, 6BPase is also a crucial regulating enzyme in glycolysis and enhances the glucose level in the diabetic state. Hence, both enzymes were found with poor recognition of insulin and released highly in diabetic rats. But in oral administered WS and control, drug-treated experimental rats show high enzymes in the initial days and at the end of medication brought to an average level like control. So, this was again proved or agreed with previously published results and our plant is good in maintaining diabetes-like control drugs.³⁵ Another essential enzyme of carbohydrate metabolism is glucose 6 phosphate dehydrogenases (G6PDH), from HMP shunt as an alternative pathway of the TCA cycle of amphibolic role.³⁸ Our study reveals that increased G6PDH was found in the oral treated WS experimental group than in diabetic rats. Also, HMP shunt-producing NADPH is utilized to manage glutathione coenzyme activity as a scavenging activity. This is also one of the reasons is that high G6PDH is enough to scavenge beta cells of insulin secretion and maintain the enzyme activity was proved by our experimental groups.

All the enzymes and the metabolism took place in the liver, and its function was declared by markers such as ALT and AST of protein catabolic enzymes. Impaired or altered AST and ALT levels imply and are associated with improper insulin transport mechanisms in middle-aged people.³⁹ A recent report reveals that poor insulin enhances the transaminases enzymes of both AST and ALT will trigger the protein catabolism directed gluconeogenesis results in a hyperglycemic condition in both alcohol and diabetic condition.³⁷ This confirms that liver markers can also be influenced by poor insulin become worse in diabetic cases. The present study oral administered WS experimental groups was increased in initial and at the end of treatment tend to turn into maintaining liver markers than STZ induced diabetic rats were shown in histopathology of liver cells. In an uncontrolled diabetic condition, the excess of glucose molecules will be accumulated in the glomerulus, and the excretion mechanism of the kidney become worse.⁴⁰

Here in the present study, STZ induced diabetic rats to show higher elevation of urea, uric acid, and creatinine than normal levels, denoting diabetes was in uncontrolled than control. But, in group 6 of WS, the oral administered case was maintained, which was confirmed by comparing normal control with the drug control group. This again proved that oral administered WS is capable of product kidney from an excessive load of excretion and its damage was shown in kidney histopathological and its cross-sectional image view is clear than diabetic rats.

After protein and carbohydrate metabolism, lipid and its classes play a significant role in controlling diabetes. Low insulin secretion immediately impairs the triglycerides and phospholipids degradation through gluconeogenesis and synthesis glucose become hyperglycemic condition.⁴¹ Total cholesterol is also a vital lipid molecule that is mediated by SREBP (sterol regulatory mediated element-binding protein) enhance the high cholesterol (TC) level was triggered by low insulin secretion. Also, the TG and total PL are essential parameters for conforming to diabetes and HbA1c. The present study estimated TC, PL, TG, and LDL as high and low HDL in diabetic rats than control. Although in oral administered WS experimental groups have high HDL and other lipid parameters was found with almost standard control drug than diabetic rats. Once again, lipid parameters confirm that our oral administered WS and its action on controlling mechanism was better than control.

Enzymatic, non-enzymatic antioxidants serve to produce the cells and tissues from cellular injuries caused by imbalanced free radicals. After that, free radicals, homeostatic fluid balance, and molecular equivalents play a significant role in maintaining cell membranes from electrical shock. Not only protecting membranes, supplying reducing equivalents to balance the coenzymes during ATP production by TCA cycle and HMP shunt.⁴² Superoxide dismutase, catalase, alpha-tocopherol, and reduced glutathione were lower in diabetic rats than in oral administered WS experimental rats. Finally, our oral administered WS drug shows its better effect on maintaining molecular reducing equivalents was known by homeostatic mechanism results in increased antioxidant activity.

Conclusion

In a nutshell, our study has claimed that treatment of STZ induced diabetic rats by oral administered Wogonoside was good anti-diabetic medicine, evidenced by triggered insulin secretion from degenerated beta cells results in controlled plasma glucose level with kidney liver markers and scavenging mechanisms.

Supplemental Material

sj-docx-1-npx-10.1177_1934578X251372861 - Supplemental material for Effect of Wogonoside in Streptozotocin Induced Oxidative Stress and Inflammation in Diabetic Rats

Supplemental material, sj-docx-1-npx-10.1177_1934578X251372861 for Effect of Wogonoside in Streptozotocin Induced Oxidative Stress and Inflammation in Diabetic Rats by Yanfang Guo, Yanbo Liu, Zhongqiang Yuan and Hongxia Shen in Natural Product Communications

Supplemental Material

sj-docx-2-npx-10.1177_1934578X251372861 - Supplemental material for Effect of Wogonoside in Streptozotocin Induced Oxidative Stress and Inflammation in Diabetic Rats

Supplemental material, sj-docx-2-npx-10.1177_1934578X251372861 for Effect of Wogonoside in Streptozotocin Induced Oxidative Stress and Inflammation in Diabetic Rats by Yanfang Guo, Yanbo Liu, Zhongqiang Yuan and Hongxia Shen in Natural Product Communications

Footnotes

ORCID iD

Hongxia Shen

Funding

The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: Handan Science and Technology research and development program (23422083323). Project Name:Platelet-rich plasma and adipose vascular components combined in the treatment of diabetic foot.

Declaration of Conflicting Interests

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

Supplemental Material

Supplemental material for this article is available online.

References

Saeedi

Petersohn

Salpea

, et al. Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Results from the International Diabetes Federation Diabetes Atlas. Diabetes Res Clin Pract. 2019;157:107843.

Daryabor

Atashzar

Kabelitz

Meri

Kalantar

. The effects of type 2 diabetes mellitus on organ metabolism and the immune system. Front Immunol. 2020;11:1582.

Rui

. Energy metabolism in the liver. Compr Physiol. 2014;4(1):177–197.

Loria

Lonardo

Anania

. Liver and diabetes. A vicious circle. Hepatol Res. 2013;43(1):51–64.

Han

H-S

Kang

Kim

Choi

Koo

S-H

. Regulation of glucose metabolism from a liver-centric perspective. Exp Mol Med. 2016;48(3):e218–e218.

Chaudhury

Duvoor

Reddy Dendi

, et al. Clinical review of antidiabetic drugs: implications for type 2 diabetes mellitus management. Front Endocrinol (Lausanne). 2017;8:6–6.

Colberg

Sigal

Yardley

, et al. Physical activity/exercise and diabetes: a position statement of the American Diabetes Association. Diabetes Care. 2016;39(11):2065.

Hassan

Ullah

. Antibacterial and antifungal activities of the medicinal plant veronica biloba. J Chem. 2019;2019:5264943.

Mariadoss

VAA

Vinyagam

Rajamanickam

Sankaran

Venkatesan

David

. Pharmacological aspects and potential use of phloretin: A systemic review. Mini reviews in medicinal chemistry. Mini-rev Med Chem. 2019;19(13):1060–1067.

10.

Chassagne

Samarakoon

Porras

, et al. A systematic review of plants with antibacterial activities: A taxonomic and phylogenetic perspective. Frontiers in pharmacology. Front Pharmacol. 2021;11(2069).

11.

Kim

D-I

Lee

S-H

Choi

J-H

Lillehoj

M-H

Lee

G-S

. The butanol fraction of Eclipta prostrata (Linn) effectively reduces serum lipid levels and improves antioxidant activities in CD rats. Nutr Res. 2008;28(8):550–554.

12.

Talchai

Xuan

Lin

Sussel

Accili

. Pancreatic β cell dedifferentiation as a mechanism of diabetic β cell failure. Cell. 2012;150(6):1223–1234.

13.

Babaei-Balderlou

Zare

. F.Melatonin improves spatial navigation memory in male diabetic rats. aculty of Veterinary Medicine. Urmia University; 2012: 187.

14.

MacDonald

Gapinski

. A rapid ELISA for measuring insulin in a large number of research samples. Metabolism. 1989;38(5):450–452.

15.

Zimmet

Shaw

Alberti

. Preventing type 2 diabetes and the dysmetabolic syndrome in the real world: a realistic view.Diabetic Med. 2003;20(9):693–702.

16.

Pouwer

Kupper

Adriaanse

. Does emotional stress cause type 2 diabetes mellitus? A review from the European Depression in Diabetes (EDID) Research Consortium. Discov Med. 2010;9(45):112–118.

17.

LeRoith

Biessels

Braithwaite

, et al.Tratamiento de la diabetes en adultos mayores: Una guía de práctica clínica de la Endocrine Society de Endocrinología J Clin Endocrinol Metab. 2019;104(5):1520–1574.

18.

Ferzacca

. Med Anthropol Q. Diabetes Cul. 2000;14(1):28–50.

19.

Forbes

Cooper

. Mechanisms of diabetic complications. Physiol Rev. 2013;93(1):137–188.

20.

Wang-Fischer

Garyantes

. Improving the reliability and utility of streptozotocin‐induced rat diabetic model. J Diabetes Res. 2018;2018:8054073.

21.

Noshahr

Salmani

Khajavi Rad

Sahebkar

. Animal models of diabetes‐associated renal injury. J Diabetes Res. 2020;2020.

22.

Uddin

Rahman

Ahmed

Rana

Akter

Chowdhury

AMA

. Eclipta alba. Int J Biol Med Res. 2010;1(4):341–346.

23.

Duncan

Perri

Theriaque

Hutson

Eckel

Stacpoole

. Exercise training, without weight loss, increases insulin sensitivity and postheparin plasma lipase activity in previously sedentary adults. Diabetes Care. 2003;26(3):557–562.

24.

Nilsson

Johansson-Boll

Björck

. Increased gut hormones and insulin sensitivity index following a 3-d intervention with a barley kernel-based product: a randomised cross-over study in healthy middle-aged subjects. Br J Nutr. 2015;114(6):899–907.

25.

Nuttall

Gannon

. Plasma glucose and insulin response to macronutrients in nondiabetic and NIDDM subjects. Diabetes Care. 1991;14(9):824–838.

26.

Canfora

Jocken

Blaak

. Short-chain fatty acids in control of body weight and insulin sensitivity. Nat Rev Endocrinol. 2015;11(10):577–591.

27.

James

Collins

Logan

Murphy

. Mitochondrial oxidative stress and the metabolic syndrome. Trends in endocrinology & metabolism. Trends Endocrinol Metab. 2012;23(9):429–434.

28.

Meliani

Dib

MEA

Allali

Tabti

. Hypoglycaemic effect of Berberis vulgaris L. in normal and streptozotocin-induced diabetic rats. Asian Pac J Trop Biomed. 2011;1(6):468–471.

29.

Sasidharan

Sumathi

Jegathambigai

Latha

. Antihyperglycaemic effects of ethanol extracts of Carica papaya and Pandanus amaryfollius leaf in streptozotocin-induced diabetic mice. Nat Prod Res. 2011;25(20):1982–1987.

30.

Jaiswal

Tatke

Gabhe

Vaidya

. Antidiabetic activity of extracts of Anacardium occidentale Linn. leaves on n-streptozotocin diabetic rats. J Tradit Complement Med. 2017;7(4):421–427.

31.

Szabat

Page

Panzhinskiy

, et al. Reduced insulin production relieves endoplasmic reticulum stress and induces β cell proliferation. Cell Metab. 2016;23(1):179–193.

32.

Murray

Lygate

Cole

, et al. Insulin resistance, abnormal energy metabolism and increased ischemic damage in the chronically infarcted rat heart. Cardiovasc Res. 2006;71(1):149–157.

33.

Matschinsky

Wilson

. The central role of glucokinase in glucose homeostasis: a perspective 50 years after demonstrating the presence of the enzyme in islets of Langerhans. Front Physiol. 2019;10:148.

34.

Nguyen

Leray

Diez

, et al. Liver lipid metabolism. J Anim Physiol Anim Nutr. 2008;92(3):272–283.

35.

Akhtar

Khan

Khaliq

. Effects of Euphorbia prostrata and Fumaria parviflora in normoglycaemic and alloxan-treated hyperglycaemic rabbits. Planta Med. 1984;50(02):138–142.

36.

Badoiu

Miricescu

Stanescu-Spinu

I-I

, et al.

Glucose metabolism in burns—what happens?

Int J Mol Sci. 2021;22(10):5159.

37.

Manimegalai

Mahboob

Al-Ghanim

, et al. Down-regulation of hepatic G-6-Pase expression in hyperglycemic rats: Intervention with biogenic gold nanoconjugate. Saudi J Biol Sci. 2020;27(12):3334–3341.

38.

Bakar

MHA

Sarmidi

Cheng

K-K

, et al. Metabolomics–the complementary field in systems biology: a review on obesity and type 2 diabetes. Mol BioSyst. 2015;11(7):1742–1774.

39.

De Bock

Derraik

Brennan

, et al. Olive (Olea europaea L.) leaf polyphenols improve insulin sensitivity in middle-aged overweight men: a randomized, placebo-controlled, crossover trial. PloS One. 2013;8(3):e57622.

40.

Mora-Fernández

Domínguez-Pimentel

de Fuentes

Górriz

Martínez-Castelao

Navarro-González

. Diabetic kidney disease: from physiology to therapeutics. J Physiol. 2014;592(18):3997–4012.

41.

Czech

. Insulin action and resistance in obesity and type 2 diabetes. Nat Med. 2017;23(7):804–814.

42.

Lobo

Patil

Phatak

Chandra

. Free radicals, antioxidants and functional foods: impact on human health. Pharmacogn Rev. 2010;4(8):118–126.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

0.03 MB

4.54 MB