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Abstract

The aim of the present study was to determine the protective and therapeutic effects of linalool (LIN) against doxorubicin (DOX)-induced cardiotoxicity in rats histologically and biochemically. In experiments, 64 male Wistar albino rats were randomly divided into eight groups (n = 8). These groups were control (C) (0.9% saline solution), DOX (20 mg/kg DOX), LIN50 (50 mg/kg LIN), LIN100 (100 mg/kg LIN), DOX + LIN50 (20 mg/kg DOX and 50 mg/kg LIN), DOX + LIN100 (20 mg/kg DOX and 100 mg/kg LIN), LIN50 + DOX (50 mg/kg LIN and 20 mg/kg DOX), and LIN100 + DOX (100 mg/kg LIN and 20 mg/kg DOX). It was determined that necrosis and extensive inflammatory cell infiltration were observed in the DOX group. It was determined that histopathological changes significantly decreased in groups treated with LIN after DOX administration. While the caspase-3 immunostaining was highly evident in DOX group apoptotic cells (p < 0.001, for all), the intensity of caspase-3 immunostaining in the treatment groups decreased (p < 0.05). While DOX administration resulted in a significant increase in malondialdehyde (MDA) levels and plasma Creatine kinase (CK) and lactate dehydrogenase (LDH) levels in cardiac tissue when compared to the C groups, it was observed that DOX + LIN administration led to a significant decrease in MDA, plasma CK and LDH levels and a significant increase in glutathione (GSH), superoxide dismutase, and catalase enzyme levels. Finally, it was concluded that DOX led to heavy cardiotoxicity and DOX + LIN administration could remove cardiomyopathy symptoms.

Keywords

Doxorubicin linalool cardiotoxicity MDA caspase-3 oxidative stress

Introduction

Doxorubicin (DOX) is an antineoplastic agent in the anthracycline group. It was demonstrated in the early 1960s that DOX, an antitumoral antibiotic, was highly effective on various malignancies.¹ DOX is one of the most effective cytotoxic drugs against solid tumors such as ovarian, breast, gastrointestinal, and Wilms’ tumors and hematologic malignant tumors such as Hodgkin and non-Hodgkin lymphoma and pediatric leukemia.² However, the drug has serious side effects that lead to congestive heart failure.¹

The pathogenesis of DOX-induced cardiotoxicity was not fully explained and due to the variations in histopathological findings, it was suggested that several factors could be active.^1,3 Cardiotoxicity is determined by cardiac exposure to anthracyclines and to more toxic secondary alcohol metabolites that are formed inside cardiomyocytes or diffuse from the bloodstream.⁴ The present study findings demonstrated that free radical formation, the decrease in antioxidant enzymes, and the increase in lipid peroxidation could play a role in the pathogenesis of DOX-induced cardiotoxicity.⁵ It was demonstrated that free radical–induced lipid peroxidation products such as malondialdehyde (MDA) contributed to the mechanism.^1,3,6 It was also demonstrated that it reduces antioxidant enzymes such as glutathione peroxidase, superoxide dismutase (SOD), and catalase (CAT), leading to cardiotoxicity.^1,3 The findings that free radicals and antioxidant enzymes play a role in the pathogenesis of DOX-induced cardiotoxicity lead to antioxidant therapy studies.⁵

Linalool (LIN) is a natural monoterpene found in essential oils of plants such as coriander, basil, and mint.⁷ This aromatic plant has several pharmacological effects including antimicrobial, antibacterial, antiviral, anti-inflammatory, analgesic, and anesthetic effects.^8
–10 LIN, a competitive antagonist of N-methyl d-aspartate (NMDA) receptors, produces antinociception via brain opioids when it inhibits NMDA receptor activity.^11,12 In addition, LIN weakly inhibits cholinesterase activity in vitro¹³ and mediates antinociceptive effect in the rat spinal cord via muscarinic neurotransmission.¹⁴ The antihyperglycemic, hypolipidemic, and antioxidant properties of LIN, its therapeutic effect on kidney function, and its preventive effect on proteinuria were evidenced.⁸ It was reported that certain plant-derived monoterpenes inhibited and halted tumor growth, and recent studies have focused on the chemopreventive/chemotherapeutic potential of LIN.¹⁰ The concentration of LIN in plasma has been reported to reach about 10 min at the first minor peak, the second major peak occurred in about 40 min, and then the drug concentration in the plasma decreased rapidly.¹⁵

To the best of our knowledge, there is no literature on the effects of LIN on cardiotoxicity. The aim of the present study was to determine the protective and therapeutic effects of LIN, an antioxidant, on DOX-induced cardiotoxicity in rats using histopathologic and biochemical methods.

Methods

Preparations of the medicine

For DOX 50 mg I.V. and infusion, the vial that included lyophilized powder and the solvent vial were procured from Kocak Corporation (Istanbul, Turkey). The lyophilized powder solvent was dissolved with injection water and prepared for 20 mg/kg intraperitoneal (i.p.) injection.^16
–18 Ninety seven percent LIN was procured from Sigma Aldrich Corporation (Lot # STBG7505, St. Louis, Missouri, USA). LIN was dissolved in 0.9% saline solution (SS) and prepared for 50 mg/kg and 100 mg/kg i.p. injection.^19

–22

Experimental animals

The study was conducted in İnönü University Experimental Animal Production and Research Center with the approval of Inonu University Experimental Animal Ethics Committee (2016/A-64). Sixty four Wistar albino rats weighing 250–300 g, aged 6–8 weeks were used. The rats were kept in special, automatically air-conditioned rooms with a constant temperature of 21° ± 2°C, 12 h of light and 12 h of darkness conditions. The base of the cages was covered with chip with the lowest risk of parasite infection, and the cage was cleaned regularly. During the experiment, the rats were provided with standard pellet diet and water ad libitum.

Experimental protocol

The rats were randomly divided into eight groups with eight rats in each group in the present study and treated as follows:

Control (C), 0.9% SS i.p., for 5 days

DOX, 20 mg/kg i.p., single dose on first day

LIN50, 50 mg/kg i.p., for 5 days

LIN100, 100 mg/kg i.p., for 5 days

DOX + LIN50, 20 mg/kg i.p., single dose on first day + 50 mg/kg for 5 days, respectively.

DOX + LIN100, 20 mg/kg i.p., single dose on first day + 100 mg/kg for 5 days, respectively.

LIN50 + DOX, 50 mg/kg for 5 days + 20 mg/kg i.p., single dose on fifth day, respectively.

LIN100 + DOX, 100 mg/kg for 5 days + 20 mg/kg i.p., single dose on fifth day, respectively.

One milliliter of LIN dissolved in 50 ml of saline, freshly every day. All applications were 1 ml for 50 mg/kg, 2 ml for 100 mg/kg and by i.p. and repeated for 5 days at the same hour. I.p. injections were prepared in two separate injectors for groups 5, 6, 7, and 8 and administered separately. After 5 days of treatment, all rats were euthanized under ketamine/xylazine anesthesia on the sixth day. The blood was collected from the heart into heparinized tubes and heart tissue samples were collected.

Histopathological analysis

The tissues were placed in 10% neutral buffered formalin (NBF) for fixation. The samples were then cut into 3–4-mm long pieces and placed in plastic tissue monitoring cartridges and fixed in NBF for 48 h. When the fixation process was completed, the pieces were washed in running tap water for 24 h. They were then dehydrated in alcohol with increasing levels (70%, 80%, 96%, and 100%), and they were made pellucid in xylene and embedded in paraffin. Five-micrometer sections were incised from paraffin blocks and transferred into gelatin-coated slides for light microscopy and immunohistochemical examinations. The slides were kept in a 37°C oven for 2 h to increase the adhesion of the tissue to the slides. Hematoxylin–eosin staining method was used to observe the general histological structure of the sections. Cardiac damage was determined semiquantitatively based on the degree and extent of histopathological variations. Accordingly, they were examined in 10 different areas to determine hemorrhage, necrosis, infiltration, and vacuolization. They were scored based on the severity of damage: 0 (no change), 1 (mild), 2 (moderate), and 3 (severe).²³ It was determined that the maximum damage score was 12. The samples were scored with Nikon Eclipse 80i light microscope and pictured with Nikon image analysis system (Nikon, Tokyo, Japan).

Immunohistochemical analysis

Immunohistochemical staining was conducted with caspase-3 antibody (ab13847; Abcam, Kimera, Turkey). Caspase-3 immunoreaction was examined semiquantitatively with Nikon Eclipse 80i light microscope. To determine the staining intensity, 10 areas in each section were examined with X20 magnification, and they were scored as (0) no staining, (1) poor staining, (2) moderate staining, and (3) severe staining. Photographs were taken with Nikon image analysis system.

Biochemical analysis

On the day of analysis, the tissues were removed from the freezer and weighed. Phosphate buffer was added to produce 10% homogenate and the tissues were homogenized in ice at 12,000 r/min for 1–2 min. MDA level was measured using the homogenate. The remaining homogenate was centrifuged at 600 × g at +4°C for 30 min to obtain the supernatant. CAT and SOD activities and glutathione (GSH) and protein levels were studied with the supernatant.

MDA analysis was conducted with the method described by Ohkawa et al.²⁴ The MDA level was measured at 535 nm with a spectrophotometer (T80 UV/VIS Spectrometer; PG Instruments Ltd, Leicestershire, UK). n-Butanol was used as the blind and tetramethoxypropane was used as the standard. The results were expressed in nanomole/gram wet tissue.

The GSH level was measured based on the method described by Ellman.²⁵ The reduced GSH was determined by measuring the intensity of the color obtained at 410 nm using a spectrophotometer. Distilled water was used as the blind. The results were expressed in nanomole/gram wet tissue.

The tissue protein content was studied with the method proposed by Lowry et al.²⁶ and used to calculate the enzyme activity.

SOD activity was measured by the method reported by Sun et al.²⁷ The absorbance of the formazan at 560 nm was used to calculate the SOD activity. Distilled water was used as the blind. SOD activity was calculated as U/g protein.

CAT activity was measured with the method reported by Aebi.²⁸ Since the separation of H₂O₂ into water and oxygen by the CAT enzyme in the supernatant leads to a decrease in absorbance at 240 nm, the reduction in absorption was recorded for 1 min to measure the enzyme activity. CAT activity was calculated as K/g protein.

The blood samples that were transferred to heparinized tubes were centrifuged at 600 × g for 4 min at +4°C to obtain the plasma, which was used to determine CK and LDH levels with Architect c1600 automated analysis kits (Abbott, Abbott Park, Illinois, USA).

Statistical analysis

Statistical analyses were conducted with SPSS (Windows version 14.0) software. All results were expressed as arithmetic mean ± standard error of the mean (SEM). It was determined that the measurable variables in all groups did not demonstrate normal distribution based on Shapiro–Wilk normality test (p > 0.05). Thus, Kruskal–Wallis variance analysis was used for the general comparison of the groups based on all variables, while Mann–Whitney U test was used for the pairwise comparison of the groups. The value of p < 0.05 was considered statistically significant.

Results

Histopathological results

C, LIN50, and LIN100 group heart tissues were in normal histological appearance. However, massive hemorrhagic areas, vacuolization, pyknotic nucleus, necrosis, and diffuse inflammatory cell infiltrations were observed in the DOX group (Figure 1).

Figure 1.

Heart histopathology photomicrograph for the C and DOX groups. (a) C group; heart tissues in normal histological appearance, H-E; ×40. (b–f) DOX group. (b) Diffuse inflammatory cell infiltrations in DOX-administered heart tissue, H-E; ×40. (c) Pyknotic nucleus and necrosis, H-E; ×40. (d) Massive hemorrhagic areas, H-E; ×10. (e) Myocardial damage with vacuolization, H-E; ×40. (g) LIN50 group, H-E; ×40. (h) LIN100 group, H-E; ×40. LIN groups, almost normal myocardial appearance. C: control; DOX: doxorubicin; H-E: hematoxylin–eosin; LIN: linalool.

The mean histopathological damage score in the DOX group was 8.37 ± 0.32. There was a statistically significant increase in the damage score when compared to the C, LIN50, and LIN100 groups (p < 0.001, for all). In DOX + LIN50 and DOX + LIN100 groups, histopathological changes were significantly reduced and the damage was prevented by LIN administration in LIN50 + DOX and LIN100 + DOX groups (Figure 2).

Figure 2.

The photomicrographs of the heart where the effect of LIN on DOX-induced myocardial damage is observed. (a) LIN50 + DOX group, H-E; ×40. (b) LIN100 + DOX group, H-E; ×40. (c) DOX + LIN50 group, H-E; ×40. (d) DOX + LIN100 group, H-E; ×40. Histopathological changes significantly decreased with the effect of LIN in all groups. DOX: doxorubicin; H-E: hematoxylin–eosin; LIN: linalool.

Histopathological comparison of the DOX group and all treatment groups revealed statistically significant differences (p < 0.05). There was a statistically significant difference between the treatment groups (LIN50 and LIN100) as well (p < 0.05). It was found that the impact of the LIN100 group was more therapeutic and preventive. The histopathological total damage score for each group is presented in Table 1.

Table 1.

The mean histopathological damage score and caspase-3 immunostaining score of all groups.^a

Groups	MHDS	Caspase-3
C	0.12 ± 0.12	0.12 ± 0.12
DOX	8.37 ± 0.32^b	1.62 ± 0.26^b
LIN50	0.12 ± 0.12^c	0.12 ± 0.12^c
LIN100	0.12 ± 0.12^c	0.12 ± 0.12^c
DOX + LIN50	6.75 ± 0.55^b,d,e	1.12 ± 0.22^i,k
DOX + LIN100	4.62 ± 0.41^b,c,e,f	0.50 ± 0.26^d
LIN50 + DOX	5.62 ± 0.32^b,c,e	0.62 ± 0.26^d
LIN100 + DOX	3.37 ± 0.53^b,e,g,h	0.37 ± 0.18^j,l

SE: standard error; C: control; DOX: doxorubicin; LIN: linalool; MHDS: mean histopathological damage score.

^aData are expressed mean ± SE of eight animals.

^bp < 0.001 versus C.

^cp < 0.001 versus DOX.

^dp < 0.05 versus DOX.

^ep < 0.001 versus LIN50 and LIN100.

^fp < 0.01 versus DOX + LIN50.

^gp < 0.005 versus DOX + LIN50.

^hp < 0.01 versus LIN50 + DOX.

ⁱp < 0.005 versus C.

^jp < 0.005 versus DOX.

^kp < 0.005 versus LIN50 and LIN100.

^lp < 0.05 versus DOX + LIN50.

Immunohistochemical results

The caspase-3 immunostaining of the C, LIN50, and LIN100 groups was characterized by very weakly stained cells. There was no statistically significant difference between the groups (p > 0.05). However, when the DOX group was compared to C, LIN50, and LIN100 groups, it was determined that caspase-3 immunostaining in apoptotic cells was imminent (p < 0.001, for all). Compared to the DOX group, the intensity of caspase-3 immunostaining decreased in treatment groups (p < 0.05; Figure 3). The caspase-3 immunostaining scores are presented in Table 1.

Figure 3.

Caspase-3 IHC staining for apoptosis in the heart tissue. C, LIN50, and LIN100 groups were similar. In the DOX group, apoptosis was most evident. The intensity of caspase-3 was reduced by linalool in treatment groups. (a) C group, (b) DOX group, (c) LIN50 group, (d) LIN100 group, (e) LIN50 + DOX group, (f) LIN100 + DOX group, (g) DOX + LIN50 group, (h) DOX + LIN100 group, caspase-3; ×20. C: control; DOX: doxorubicin; LIN: linalool; IHC: immunohistochemistry.

Biochemical results

DOX administration resulted in significant increases in cardiac tissue MDA levels when compared to the C and LIN groups (p < 0.05). Both LIN doses (50 mg and 100 mg) administered with DOX resulted in a significant decrease when compared to the DOX group (p < 0.05). However, no significant difference was found between the administration of two LIN doses (50 mg and 100 mg) in treatment groups. When the DOX administration was conducted after LIN supplement, it was found that the MDA level was significantly lower when compared to the DOX group (p < 0.05). There was no significant difference between LIN50 and LIN100 applications. Similarly, there was no significant difference between the LIN administration (DOX + LIN50 and DOX + LIN100) after DOX administration and DOX administration after LIN administration (LIN50 + DOX and LIN100 + DOX).

GSH levels decreased significantly after DOX administration (p < 0.05), whereas LIN applications (50 mg and 100 mg) led to a significant increase when compared to C and DOX groups (p < 0.05). LIN50 and LIN100 administration after DOX application and DOX application after LIN50 and LIN100 administration led to a GSH level similar to the C group and a significant increase when compared to the DOX group (p < 0.05).

DOX application resulted in significant decreases in SOD and CAT enzyme activities when compared to the C group (p < 0.05). SOD and CAT activities significantly increased when compared to the C and DOX groups with 50 mg and 100 mg LIN doses (p < 0.05). Furthermore, 50 mg and 100 mg LIN administration after DOX application resulted in significant increases in enzyme activities when compared to the DOX group (p < 0.05); however, no significant difference was found between the two LIN doses. When DOX was administered after LIN treatment (LIN50 + DOX and LIN100 + DOX), there was a significant increase in enzyme activities when compared to the DOX group (p < 0.05). In addition, when we compare LIN 100 + DOX group and LIN 50 + DOX group, it was found that enzyme activity decreased significantly (p < 0.05). Furthermore, DOX application after 100 mg LIN administration resulted in a significant increase when compared to DOX + LIN50 and DOX + LIN100 groups (p < 0.05). Cardiac tissue oxidant–antioxidant parameters are presented in Table 2.

Table 2.

The levels of tissue oxidant–antioxidant parameters of all groups.^a

Groups	MDA (nmol/g wet tissue)	GSH (nmol/g wet tissue)	SOD (U/g protein)	CAT (K/g protein)
C	355.87 ± 20.25	527.87 ± 10.06	27.18 ± 1.21	2.74 ± 0.08
DOX	643.75 ± 22.39^b	471.87 ± 13.38^d	18.83 ± 1.24^d	1.80 ± 0.18^d
LIN50	310.75 ± 19.36^c	585.62 ± 16.55^f,c	33.01 ± 1.24^b,c	3.97 ± 0.29^i,c
LIN100	286.87 ± 9.75^c,d	617.75 ± 17.91^c	37.72 ± 1.73^b,c	4.38 ± 0.23^b,c
DOX + LIN50	459.00 ± 35.04^d,e	502.25 ± 7.06	25.60 ± 1.05^e	3.23 ± 0.32^e
DOX + LIN100	441.50 ± 13.42^d,c	514.37 ± 9.51^g	26.82 ± 1.21^e	3.36 ± 0.26^i,c
LIN50 + DOX	458.12 ± 34.38^e	565.75 ± 31.68^h	27.76 ± 1.08^e	3.41 ± 0.25^f,c
LIN100 + DOX	403.37 ± 30.71^c	531.25 ± 16.52^g	27.27 ± 1.02^e	4.15 ± 0.15^b,c,j,k

SE: standard error; C: control; DOX: doxorubicin; LIN: linalool; MDA: malondialdehyde; SOD: superoxide dismutase; GSH: glutathione; CAT: catalase.

^aData are expressed mean ± SE of eight animals.

^bp < 0.001 versus C.

^cp < 0.001 versus DOX.

^dp < 0.005 versus C.

^ep < 0.005 versus DOX.

^fp < 0.01 versus C.

^gp < 0.05 versus DOX.

^hp < 0.01 versus DOX.

ⁱp < 0.05 versus C.

^jp < 0.05 versus DOX + LIN50.

^kp < 0.05 versus DOX + LIN100.

Due to the cardiotoxicity induced by DOX application, plasma CK and LDH levels significantly increased when compared to the C and LIN groups (LIN50 and LIN100) (p < 0.05). LIN administration (DOX + LIN50 and DOX + LIN100) after DOX application and DOX application after LIN administration (LIN50 + DOX and LIN100 + DOX) resulted in a significant decrease in enzyme levels when compared to the DOX group (p < 0.05). There was no significant difference between DOX + LIN50 and LIN50 + DOX. Plasma CK and LDH levels are presented in Table 3.

Table 3.

The levels of blood biochemical parameters of all groups.^a

Groups	CK (U/L)	LDH (U/L)
C	114.37 ± 9.74	266.50 ± 15.337
DOX	370.37 ± 36.79^b	619.62 ± 55.89^b
LIN50	110.12 ± 9.97^c	253.37 ± 22.42^c
LIN100	87.12 ± 6.92^c	242.25 ± 15.67^c
DOX + LIN50	273.12 ± 21.89^d	521.75 ± 56.84^b
DOX + LIN100	203.12 ± 24.19^e	457.12 ± 36.17^b,d
LIN50 + DOX	221.00 ± 27.04^d	482.37 ± 32.95^b
LIN100 + DOX	183.12 ± 25.02^f	414.62 ± 39.37^g,d

SE: standard error; C: control; DOX: doxorubicin; LIN: linalool; CK: creatine kinase; LDH: lactate dehydrogenase.

^aData are expressed mean ± SE of eight animals.

^bp < 0.001 versus C.

^cp < 0.001 versus DOX.

^dp < 0.05 versus DOX.

^ep < 0.01 versus DOX.

^fp < 0.005 versus DOX.

^gp < 0.01 versus C.

Discussion

Most cytostatic drugs used in chemotherapy have low selectivity for cancer cells and lead to toxicity in normal cells, particularly in bone marrow, intestine and gonad tissues, as well as organs such as the heart, liver, and kidney.² Cardiotoxic side effect of DOX, an active antitumor agent, prevents the maximum benefits of the antitumor effect due to its cardiotoxic effect and thus, prevention of cardiotoxicity became an important research field.²⁹

Studies on prevention of DOX-induced cardiotoxicity are mostly experimental and in studies conducted on mice, rats, rabbits, pigs and dogs, the clinical and morphological properties of DOX-induced cardiotoxicity were similar to that in humans.²⁹ In the present study, rat model was preferred, and young rats were selected to reflect the cardiotoxic action of DOX in childhood. The DOX dose was administered at the total cumulative dose (20 mg/kg) determined in animal models.^16
–18

The histopathological findings of the present study were consistent with the findings in the literature.^2,3,30
–32 Both the administration of LIN before the DOX and administration of LIN after DOX significantly reduced the histopathological damage score, indicating that LIN is both therapeutic and preventive. In a study conducted by Mehri et al.²⁰ on LIN neurotoxicity, the protective effect of 12.5 mg LIN was reported. However, they reported no therapeutic effect. The present study, both 50 mg and 100 mg doses, exhibited both preventive and therapeutic effects on cardiotoxicity.

Caspase-3 is a mediator of the mitochondrial apoptotic pathway and is known to be an indicator of oxidative stress–induced necrosis.^2,33 It was observed that caspase-3 immunostaining in the DOX group apoptotic cells was quite significant, while the staining was very weak in LIN-treated group apoptotic cells. Thus, it is suggested that immunohistochemical results supported histopathological findings.

Although the mechanism of DOX-induced toxicity is not fully known, it is considered that oxidative stress, apoptosis, and inflammation play a role in the mechanism.² In in vitro cellular and in vivo animal models, it was reported that DOX-induced myocardial injury was reduced by increasing endogenous antioxidant enzyme activities.^34

–37 In the present study, to determine the DOX-induced oxidative stress in cardiac tissues, MDA and GSH levels and SOD and CAT enzyme activities were measured and the regulatory effect of LIN, a natural antioxidant, was investigated.

GSH is the major antioxidant that protects cardiomyocytes against free oxygen radicals.³⁸ In the present study, the heart tissue GSH level decreased only in the DOX-administered group, which confirmed the primary role of GSH reduction in the pathogenesis of DOX cardiotoxicity. In the DOX + LIN-administered groups, GSH levels were maintained in the present study. Histopathological reduction of cardiotoxicity was also explained by the elevated cardiac GSH levels.

In a study conducted by Demir et al.,³ the antioxidant activities of melatonin were analyzed and they reported an increase in SOD activity in the melatonin group when compared to the C group. In a study conducted by Xu et al.,³³ a significant increase in SOD activity was observed in hippocampus and cortex in 100 mg/kg LIN-administered groups. Similarly, SOD and CAT enzyme activities significantly decreased after DOX administration when compared to the C group in the present study, while in both LIN-administered groups (50 mg and 100 mg), there was a significant increase when compared to the C and DOX groups. This demonstrated the antioxidant effect of LIN in both doses.

It was considered that another mechanism related to free radical–mediated and DOX-induced cardiac injury was lipid peroxidation. In the literature, several studies reported an increase in cardiac tissue MDA levels due to the pathophysiology of DOX-induced cardiotoxicity.^39
–41 In the present study, there was a significant increase in cardiac tissue MDA levels in the DOX group, while both LIN doses administered before and after DOX administration significantly decreased the MDA levels. This finding suggested that LIN exhibited antioxidant activities by inhibiting lipid peroxidation. In a study conducted by Xu et al.,²² a significant reduction in hippocampus MDA levels was observed.

Saad et al.³¹ explained the increase in cardiac enzymes such as CK and LDH with the enzyme release as a result of DOX-induced lipid peroxidation in cardiac membranes. In a study conducted by Dai et al.,⁴² plasma CK and LDH levels significantly decreased with octreotide treatment in DOX-administered rats. Similarly, in the present study, a significant increase in plasma CK and LDH levels was observed due to DOX-induced cardiotoxicity. LIN administration before and after DOX administration led to a significant decrease in enzyme levels. These biochemical results supported both the therapeutic and protective effects of LIN observed in histopathological analysis.

In conclusion, it was determined that DOX led to severe cardiotoxicity, administration of LIN with DOX reduces cardiomyopathy symptoms, and both 50 mg and 100 mg LIN administration could protect the tissues against DOX-induced cardiotoxicity and might have a therapeutic effect. There is little literature on the cardioprotective activity of LIN and there are only studies on experimental animals. However, further studies are needed to use LIN as a cardioprotective in the clinic. It is expected that the findings of this project would guide future studies by oncologists, pharmacologists, and scientists interested in alternative medicine.

Footnotes

Declaration of Conflicting Interests

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

Funding

The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was sponsored by TUBITAK 1002 (project no. 216S330) Fast Support Program.

ORCID iD

Z Oner

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The protective and therapeutic effects of linalool against doxorubicin-induced cardiotoxicity in Wistar albino rats

Abstract

Keywords

Introduction

Methods

Preparations of the medicine

Experimental animals

Experimental protocol

Histopathological analysis

Immunohistochemical analysis

Biochemical analysis

Statistical analysis

Results

Histopathological results

Immunohistochemical results

Biochemical results

Discussion

Footnotes

Declaration of Conflicting Interests

Funding

ORCID iD

References