Anti- Atherosclerotic Activity of Salvia Splendens On Triton X- 100 Induced Atherosclerosis in Experimental Animal

Abstract

Herbal plants contains several bioactive constituents may possess potential cardioprotective effects. The objective for present research work is to investigate herbal medication for atherosclerosis. The attempt is made to investigate this plant with promising effectiveness against atherosclerosis. Therefore the present study is used to evaluate anti-atherosclerotic activity of Salvia splendens in triton x -100 induced atherosclerotic rat model. Atherosclerosis were induced in male wistar rats by intraperitoneal administration of Triton X-100mg/kg body weight for 21 days. The methanolic extract of Salvia splendens leaves (100, 300 mg/kg) administered orally to Triton x- 100 treated rats for next 21 days. Rats were subjected to evaluate general parameter and biochemical parameters. Physical parameters were estimated at 1, 7, 14, 21 days. Biochemical parameters were also estimated in the same intervals by collecting the blood samples from tail vein under mild ether anesthesia. Then the rats were sacrificed on 22nd day, the heart was dissected out for histopathological studies. The elevated serum biochemical parameter such as TC (total cholesterol), LDL (low density lipoprotein), VLDL (very low density lipoprotein), HDL (high densisty lipoprotein, TG (triglyceride) and AI (atherogenic index). Triton X- 100 treated rats were reverted back approximately near to normal after the treatment of Salvia splendens leaves at a doses of 100 and 300 mg/kg. Salvia splendens leaves possess protective action against Triton X- 100 induced atherosclerosis in wistar rats which may be attributed by its antioxidant, anti-inflammatory and anticoagulant effect.

Keywords

Introduction

The research finding from the study graphical representation of TC, LDL, VLDL and TG all shows biochemical parameter significantly decrease and Graph of HDL shows significantly increase by the oral consumption of extract, especially at the prescribed dosages of 100mg/kg and 300mg/kg. This result may ultimately imply a potent activity against atherosclerosis. Such outcomes underscore the extract may be potential effect and mechanism and shows significantly increase the value of HDL which indicate that improving the biological activity.                

Results of Histopathology:

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A) Normal Control group:

Shows the cross section of the heart of the sham group stained with haematoxylin and eosin. Section of heart shows no fatty changes with normal tissuearchitecture, multiple peripheral nuclei, and regular morphology of myocardial cell membrane.

B) Inducing group Triton X 100 (100mg/kg):

Shows the cross section of the heart of the negative control group stained with haematoxylin and eosin. Section heart shows alteration in tissue architecture and focal fatty infiltration in myocardial cells.

C) Standard group (Fenofibrate 65mg/kg):

Shows the cross section of the heart of standard control group stained with haematoxylin and eosin. Section of heart shows restored tissue and myocardial architectures.

D) Treatment group MESS 100mg/kg (Methanolic extract Salvia splendens):

Shows the cross section of the heart of 100 mg/kg bw group stained with haematoxylin and eosin. Section heart shows focal fatty infiltration in myocardial cells.

E) Treatment group MESS 300mg/kg (Methanolic extract Salvia splendens):

Shows the cross section of the heart of the 300 mg/kg bw group stained with haematoxylin and eosin. Heart shows positive cardiomyocytes restoration with very mild fatty infiltration.

DISSCUSION:

The current study was designed to assess the effect of MESS against Triton X-100-induced atherosclerosis in rats. Triton X-100 is reported to block the clearance of TG-rich lipoproteins and produces acute hyperlipidemia and atherosclerosis in animal models. [16-18] Plasma TG and cholesterol levels were elevated due to Triton X-100 injection as an increase of VLDL secretion by the liver, followed by reduction of VLDL and LDL catabolism. In the present study, Triton X-100 significantly increased the serum TC, LDL-C, VLDL-C, TGs, AI and decreased HDL-C levels as also reported in a previous study. [19-20] Triton X-100 inhibits lipoprotein lipase, the enzyme responsible for hydrolyzing triglycerides in chylomicrons and very low-density lipoproteins (VLDL). This inhibition results in accumulation of triglyceride-rich lipoproteins in the plasma Blocking Hepatic Uptake of Lipoproteins. This leads to elevated levels of plasma lipids, particularly cholesterol and triglycerides. Rapid Induction of Hyperlipidemia and atherosclerosis by intraperitoneal or intravenous administration, Triton X-100 causes acute hyperlipidemia within a few hours depending on the dose and species used. Oxidative Stress and endothelial dysfunction causes excessive lipoproteins in circulation can become oxidized, contributing to oxidative stress. Oxidized LDL promotes endothelial injury, inflammation, and foam cell formation, which are key steps in atherogenesis. Triton X-100 models are typically used for acute hyperlipidemia, repeated or sustained exposure may contribute to early atherosclerotic changes, especially in combination with a high-fat diet or genetic susceptibility result atherosclerotic lesion formation (In Long-Term Use). Salvia splendens contains several bioactive constituents may possess potential cardioprotective effects. Among these, rosmarinic acid, a phenolic acid, plays a key role due to its strong antioxidant and anti-inflammatory properties. It scavenges reactive oxygen species (ROS) and helps protect the vascular endothelium from oxidative damage. Luteolin, a flavonoid, exhibits both anti-inflammatory and vasodilatory effects by inhibiting the NF-κB pathway and enhancing nitric oxide (NO) production, which contributes to improved vascular function. [21-22] Another important flavonoid, apigenin, functions as an antioxidant and anti-apoptotic agent, primarily by reducing oxidative stress in cardiomyocytes and helping to maintain cellular integrity under stress conditions. Additionally, Salvia splendens has been reported to contain salvianolic acid-like polyphenols, which are known to improve endothelial function. These compounds act by promoting endothelial nitric oxide synthase (eNOS) activity and inhibiting platelet aggregation, both of which are crucial for maintaining vascular health and preventing thrombosis. Ursolic acid, a triterpenoid also found in the plant, exhibits anti-hyperlipidemic and anti-atherosclerotic activities by lowering LDL cholesterol levels and inhibiting the formation of foam cells, which are central to plaque formation in atherosclerosis. Lastly, though less confirmed in Salvia splendens tanshinone-like diterpenes may contribute to cardioprotection by improving mitochondrial function and reducing infarct size during ischemic events. Together, these compounds highlight the potential of Salvia splendens as a source of natural agents for cardiovascular protection. In the current study, MESS was able to reverse the Triton X-100-induced levels of serum TC, LDL-C, VLDL-C, TG and decreased the level of HDL-C toward normal limits. The effects of MESS on the markers of atherosclerosis were comparable to Fenofibrate 65 mg/kg, which is mostly prescribed as a standard antihyperlipidemic and ant- atherosclerotic drug. Thus, based on the markers of atherosclerosis, it may be concluded from the present study that MESS may inhibit the synthesis of cholesterol via the mevalonate pathway, and its efficacy was mildy similar to fenofibrate.  In the present study, it was established that AI increased in Triton X-100-administered rats as reported in a previous study. AI which is a measure of the extent of atherosclerotic lesion, was significantly reduced by the oral treatment of MESS and Fenofibrate, thus findings shows that pretreatment with both doses of MESS showed decreased AI. Reactive oxygen species are responsible for causing atherosclerosis. Lipid peroxidation induced by reactive oxygen species is a key factor for atherosclerosis. In the present study, there was a significant increase in the levels of lipid peroxide in the liver tissue. MESS (100 mg/kg) significant decrease in the levels of lipid peroxide. The Triton X-100-treated animals were under severe stress as it was shown by the reduction in liver enzymes and markers of oxidative stress. One study reported that fenofibrate reduced the formation of free radicals in vitro, lipid peroxides, and elevated antioxidant enzymes, thus demonstrating antioxidant effects in hyperlipedemic rat. [23-27] The levels of antioxidant enzymes, such as catalase and SOD, are decreased during cellular inflammation. These enzymes remove oxygen free radicals, thus decreasing inflammatory conditions. We observed that MESS treatment increased the SOD levels more than the control group. A similar pattern of observation with fenofibrate treatment resulted in increased SOD levels more than the vehicle control group. The reason for such effect may be the system was under severe stress, and some compensatory mechanisms may be operative to detoxify the production of reactive oxygen species. While the reason for such an effect observed by MESS is not clear, one possible explanation for an insignificant reduction could be due to a delayed decline of such antioxidant enzymes follow treatment with MESS.  Histopathological studies are the most precise evidence for the protective effect of drug as protectants. Simultaneous treatment of MESS with Triton X-100 exhibits less damage to the endothelial cells as compared with rats treated with Triton X-100 alone. The Triton X-100-treated group showed inflammatory cells of inflammation, apoptosis, and proliferation of fibrous connective tissue. Moreover, the MESS- treated group showed less proliferation of endothelial cells in histopathological examinations. MESS was able to showed beneficial effects by maintaining the normal architecture of the heart tissues, which was altered in atherosclerosis as evidenced by histopathological studies. Almost negligible damage to a few endothelial cells was observed in MESS-treated rats. The results of a histopathological study further support the results of biochemical parameters. Thus, the histopathological evidences again confirm the cardioprotective and anti- atherosclerotic effects of MESS. Further research is needed to confirm the molecular pathways and potential therapeutic use of MESS in atherosclerosis in different animal models with multiple doses.

CONCLUSION:

The above data suggest that Salvia splendens leaves extract possess weight reduction properties with no interference with inflammation. The findings of the study ensure that Salvia splendens leaves extract effectively able to reduce the serum lipids like LDL-C, TC, TGL, VLDL-C, AI and significant increase in HDL-C. It can be concluded that Salvia splendens leaves extract has a potential role in suppressing triton X induced atherosclerosis. Further investigation is needed to confirm the anti-atherosclerotic of Salvia splendens clinically. While some of these compounds are better studied in related Salvia species, their presence in S. splendens suggests that the plant may serve as a valuable natural source for developing therapeutic agents against cardiovascular diseases, particularly atherosclerosis. Further pharmacological and clinical studies are warranted to fully validate and explore its therapeutic efficacy.

FUTURE PROSPECTIVE:

Research on the anti- atherosclerotic activity of Salvia splendens is still in its early stages. Preclinical studies using animal models have shown promising results, indicating that extracts of Salvia splendens can mitigate the atherosclerotic activity of triton X 100 For example, studies have demonstrated reduction in TC, LDL, VLDL, TGL and increasing HDL parameter. Histopathological improvements in heart tissue. However, more research is needed to fully understand the mechanisms involved and to establish the efficacy and safety of Salvia splendens in humans. Future research should focus on:

• Conducting well-designed clinical trials to confirm the anti- atherosclerotic effects of Salvia splendens in patients undergoing atherosclerosis.
• Determining the optimal dosage and form of Salvia splendens for anti- atherosclerotic effects.
• Further elucidating the molecular pathways through which Salvia splendens exerts its protective effects.
• Exploring potential synergistic effects of combining Salvia splendens.with other cardioprotective agent or standard treatments to enhance overall heart protection.

CONFLICT OF INTEREST : We decleared that we have no conflict of interest

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