EVALUATION OF HEPATOPROTECTIVE, ANTIOXIDANT AND CARDIOPROTECTIVE EFFECTS OF AERIAL PART OF QUISQUALIS INDICA

LINN RATS

SUNEEL KUMAR*1, JITENDRA SINGH 2 ANUBHAV DUBEY1,

1- Department of Pharmacology, Advance Institute of Biotech and Paramedical Sciences Kanpur 209217- India.

2-Assistant Professor, Department of Pharmacolog, Pharma Dept SR Foundation Jee College, Unnao Kanpur

Corresponding Author SUNEEL KUMAR Email. Suneelkumarvk7860@gmail.com

ABSTRACT: To evaluate the Hepatoprotective, Antioxidant and Cardioprotective activity of the ethanolic extract of the leaves of Quisqualis indica against CCl4 and paracetamol induced hepatotoxicity and against doxorubicin induce cardiotoxicity in experimental rat model. The liver may be considered as the most important organ in the drug toxicity for two reasons: On one hand it is the functionally interposed between the site of absorption and the enzymatic circulation and is a major site of metabolism and elimination of foreign substances; but on the other hand these features also render it a preferred for drug toxicity. Liver is the largest organ and it is target for toxicity because of its role in clearing and metabolizing chemicals through the process called detoxification. In the present study the powdered leaves were extracted with pet ether and ethanol the resultant ethanolic extract was subjected for acute toxicity study and extract was carried out up to the dose 2000mg/kg according to OECD guidelines-425. Hepatoprotective activity and antioxidant activity of extract was studied against CCl4 and paracetamol induced hepatotoxicity in rats. Hepatoprotective Biochemical biomarkers (ALT, AST, ALP) and Antioxidant parameter (SOD) were evaluated. Cardioprotective activity of extract studied against doxorubicin induce cardiotoxicity and Lipid profile parameter (TC, TG, HDL, LDL) and cardiac biomarker (LDH, CK-MB) were studied. The activities of antioxidant enzyme were significantly increased by extract at 200 and 400 mg/kg. The Level of ALT ALP and AST also reduced to normal compared to the induce group. It also prevents the weight loss in animals. Pretreatment with Quisqualis indica extract showed reduction in blood lipid profile levels with concomitantly increase in HDL cholesterol was observed. Heart tissue injury by doxorubicin in rats was indicated by elevated level of the marker enzymes such as LDH and CK-MB. In the present study Quisqualis indica extract was found to inhibit the doxorubicin induced CK-MB and LDH release in the serum of rats. From the above all findings, it was concluded that Ethanolic extract of Quisqualis indica exhibited hepatoprotective, antioxidant and cardioprotective activity in validated animal model.

Keywords: Quisqualis indicia, Antioxidant, Carbon tetra chloride, Paracetamol, Doxorubicin.

INTRODUCTION

CVD remains the principal major cause of death in both developed and developing countries, accounting for roughly 20% of all worldwide deaths per year. In addition, hypertension, arrhythmia, coronary artery disease, MI and cardiac failure represent the leading killer of males over the age of 45 and females over the age of 65 in the United State and accounts for 750,000 deaths annually1.Cardio protection includes “All mechanisms and means that contribute to the preservation of the heart by reducing or even preventing myocardial damage”2.Although some of the considerable risk factors for CVD are not modifiable- age, sex, genetic predisposition-diet plan and lifestyle plan issue are recognized as the major modifiable risk factors3. Accordingly, current knowledge that favors the that the risk factors other than raised plasma cholesterol play an major role in the development of CVD4. Clearly, recent findings have illuminated the significant of oxidative stress, vascular inflammation, and endothelial dysfunction (as central) to the development of CVD5. Such advancements in the knowledge the disease processes have also provided new avenues to develop novel pharmaceutical and/or dietary strategies plan to cure the development of vascular diseases6.IHD is defined as acute or chronic form of cardiac disability arising from imbalance between the myocardial anoxia , the alternate term coronary artery disease is used synonymously with IHD. It is caused by disease affecting the coronary arteries. They are coronary atherosclerosis, superadded changes in coronary atherosclerosis (acute changes in chronic atheromatous plaque, coronary artery thrombosis, local platelet aggregation and coronary artery spasm) and non- atherosclerotic causes.There are two major types of ischemic manifestations:

1- Myocardial infraction2- Non-infract effects of myocardial ischemia ( angina pectoris, chronic ischaemic heart

Quisqualis indica (Combretaceae) is commonly known as “Madhumalti”. The plant is broad spread distributed ranging in all over the world especially on India, China, Philippines, Bangladesh, Myanmar and Malaysia as ornamental plant in most of the garden. Leaves with distinctive venation, oblong to elliptic, 7-15cm in length with acuminate tip and rounded base. They are quite simple and opposite. The leaves and roots extracts are reported as anthelmintic. Fresh leaves relieve flatulence. Leaves are also used application externally to treat boils and ulcers. Seeds are reported as the anthelimintic given to children to expel the worms. Leaves also used to relieve the fever. It is reported as plant roots are used to treat rheumatism. Flowers are applied to relieve headache. This plant is mostly used to treat pyrexia, diarrhea, helminthic infection, ulcer, headache, anti-inflammatory, antiseptic, and immunomodulatory activity.7

Fig.1 Quisqualis Indica Linn

MATERIALS AND METHODS

IDENTIFICATION, COLLECTION AND AUTHENTICATION OF PLANT MATERIAL:

Aerial part of Quisqualis indica were collected from herbal garden from Kanpur in the month of July and identified by Department of horticulture, Chandra Shekhar Azad Agriculture University (CSAU) Kanpur, U.P. India.

EXPERIMENTAL RODENTS

Grown-up pale cleaned individual Wistar rodents (150-250g) of both sex were used for the examination. Polypropylene limits were used to house (3 for each pen) the animal at a temperature of 28±5 °C and 12 h light/dull cycle. Hindustan Lever chow pellets were used to feed the animal and water not basic. The animals were kept fasting medium-term going before the experimentation and all of the methods used in these assessments were certified by the Institutional Animal Ethics Committee. Institutional Animal Ethics Committee (1122/PO/Re/S/07/CPCSEA, 18.08.2018) embraced all framework used in the assessment.

DRUGS AND CHEMICALS

Carban tertrachloride was gotten from Sigma Aldrich, Bangalore, India or Paracetamol (Dr. Reddy's Laboratory, Hyderabad, India) and Silymarin and Doxorubicin Akums Drugs and Pharmaceuticals, Delhi, India and various solvents were of logical assessment.

Extraction procedure:The plant materials (leaves) of Quisqualis indica L. (200 g) were dried at 40 °C for 1 week and pulverized. The powder was packed into the thimble of a Soxhlet extractor and refluxed continuously for 6 h. The solvent - either petroleum ether (PEE), ethanol (EE)) - was changed at the end of every 6 h. The solvent was removed by distillation on a Boiling water-bath at atmospheric pressure and then under reduced pressure in a rotary evaporator. The alcoholic extract was filtered, concentrated under reduced pressure to a semisolid mass and was made free from solvent. The final produced extract was weighed; percentage yield was calculated, noted and the crude extract kept in a cool place. Before administration, extract was reconstituted by dissolving in water.

METHODOLOGY

CCL4 HEPATOTOXICITY 8-9

Animals: Male Albino rat Weight: 130-150g

Groups: 5 groups were taken in which each group contained 6 rats. Animals were grouped into following groups: 14 days treatment [everyday]

Group I: Serve as a normal control Group (normal saline- Dose- 3ml/kg) Group II: Toxic control CCl4 (1ml/kg in 30% liquid paraffin solution, i.p.) Group III: Treatment control CCl4 (1ml/kg in 30% liquid paraffin solution, i.p.) + extract (200mg/kg b.w., orally)

Group IV: Treatment control CCl4 (1ml/kg in 30% liquid paraffin solution, i.p.)+ extract (400mg/kg b.w., orally)

Group V: Standard control CCl4 (1ml/kg in 30% liquid paraffin solution, i.p.)+Silymarin (100mg/kg b.w., orally) Standard drug

PROCEDURE:

The animals were kept fasted overnight on 14th day of experiment. On the next day the blood was collected from retro orbital plexus. The serum was separated by centrifugation at 3000 rpm at 20oc for 15 minutes and the biochemical parameters like ALT ALP AST SOD were estimated by using commercial kits procured from Autospan Diagnostics Pvt. Ltd., India with the aid of clinical chemistry analyzer (Lab life chem. Master). The liver and kidney in each case was dissect out, blotted of blood, washed in saline and stored in a freezer. Liver, kidney and serum were used for various biochemical estimations. Some portion of liver from each group was aseptically excused and stored in 10% formalin for histopathological studies.

EVALUATION PARAMETERS:

SOD, ALT, AST, ALP

Estimation of ALPProcedure – Serum/plasma 10 ul Working ALP reagent 1000 ul Mix well and aspirate immediately for measurement Program analyzer as per assay parameter Blank the analyzer with purified water. Read absorbance after 60 second; repeat reading after every 60 sec that is upto 180 sec at 410 nm Detect the mean absorbance change per minute (A/min)

ALP activity = A/min. 2712

Estimation of ALT

Procedure – Serum/plasma 100 ul Working ALT reagent 1000 ul Mix well and aspirate immediately for measurement Program analyzer as per assay parameter Blank the analyzer with purified water. Read absorbance after 60 second; repeat reading after every 120 sec that is upto 180 sec at 360 nm Determine the mean absorbance change per minute (A/min)

ALT activity = A/min. 1768

Estimation of AST

Procedure – Serum/plasma 100 ul Working AST reagent 1000 ul Mix well and aspirate immediately for measurement Program analyzer as per assay parameter Blank the analyzer with purified water. Read absorbance after 60 second, repeat reading after every 30 sec that is upto 120 sec at 340 nm Determine the mean absorbance change per minute (A/min)

AST activity = A/min. 1768

PARACETAMOL-INDUCED HEPATOTOXICITY IN RATS10

Animal: Albino rats of either sex Weight: 150-200g.

Group I: Normal control administered with normal saline 5ml/kg body weight for 7 days. Group II:Negative control rats received paracetamol (750mg/kg body weight, orally) for 7 days.

Group III: Administered with test extract (250mg/kg b.w., orally) and paracetamol (750mg/kg body weight, orally) for 7 days

Group IV: Administered with test extract (250mg/kg b.w., orally) paracetamol (750mg/kg body weight, orally) for 7 days

Group V: Administered standard drug silymarin (100mg/kg b.w., orally) and paracetamol (750mg/kg body weight,) for 7 days orally.

PROCEDURE:

At the end of the experiment on 8th day, blood was collected by retro-orbital puncture and was allowed to clot at room temperature. The serum was separated by centrifugation at 3000 rpm at 20oc for 15 minutes and the biochemical parameters like ALT ALP AST SOD were estimated by using commercial kits procured from Autospan Diagnostic.s Pvt. Ltd., India with the aid of clinical chemistry analyzer (Lab life chem. Master).

Estimation of ALP

Procedure – Serum/plasma 20 ul Working ALP reagent 1000 ulMix well and aspirate immediately for measurement Program analyzer as per assay parameter and blank the analyzer with purified water.

Read absorbance after 30 second, repeat reading after every 30 sec that is upto 120 sec at 405 nm Determine the mean absorbance change per minute (A/min)

ALP activity = A/min. 2712

Estimation of ALT

Procedure – Serum/plasma 100 ul Working ALT reagent 1000 ul Mix well and aspirate immediately for measurement Program analyzer as per assay parameter Blank the analyzer with purified water.Read absorbance after 60 second, repeat reading after every 30 sec that is upto 120 sec at 340 nm Determine the mean absorbance change per minute (A/min)

ALT activity = A/min. 1768

Estimation of AST

Procedure – Serum/plasma 100 ul Working AST reagent 1000 ul Mix well and aspirate immediately for measurement Program analyzer as per assay parameter Blank the analyzer with purified water.

Read absorbance after 60 second, repeat reading after every 30 sec that is upto 120 sec at 340 nm Determine the mean absorbance change per minute (A/min)

AST activity = A/min. 1768

DOXORUBICIN INDUCED MYOCARDIAL TOXICITY IN RATS: 11

Animal: Albino rats of either sex Weight: 150-200g.

Group I: Animals were administered with normal saline 5ml/kg body weight (i.p.)

Group II: Animals were administered with doxorubicin (2.5 mg/kg body weight i.p.) 6 equal injections alternatively for 2 weeks to make a total cumulative dose of 15 mg/kg body weight. Group III: animals were administered with extract (200 mg/kg b.w., orally) for 2 weeks and then alternatively with vehicle for next 2 weeks.

Group IV: Animals received extract (400 mg/kg b.w., orally, for 2 weeks) as a pretreatment followed by doxorubicin administration as in group 2.

After 36 hr of the last treatment, retro orbital plexus blood samples were obtained under light ether anesthesia using heparinized micro capillaries for the estimation of Cardiac biomarkers CPK and LDH and Total cholesterol, triglycerides, LDL.

U/L = 9683 x DA 340nm/min; Where DA = Change in absorbance.

Histopathological studies

Liver was preserved in 10% neutral buffered formalin solution for 24 h and washed with 70% ethanol. Tissues were then placed in small metal caskets, stirred by a magnetic stirrer, dehydrated

using alcohol series from 70% to 100% alcohol and embedded in paraffin using an embedding machine. Paraffin blocks were sectioned using a rotary ultra-microtome, distributed Onto glass slides and then dried overnight. Slides were observed under a light microscope after being stained with hematoxylin and eosin (H&E) dyes and mounted. Two pathologists performed the histopathologica

STATISTICAL ANALYSISThe Statistical analysis was carried out by one way analysis of variance (ANOVA) Followed by Dunant’s multiple range tests. The values are represented as Mean + SEM. Probability value at P <0.01 was considered as statistically significant

RESULT AND DISCUSSION

CCL4 HEPATOTOXICITY

After treatment with carbon tetra chloride (CCl4), rats were developed at hepatic damage observed as elevated serum levels of hepato specific enzymes like ALT, AST, ALP and SOD when compared to normal control. Pretreatment with ethanolic extract 200mg/kg showed better protection in treated rats as compared to toxic control. Silymarin (100 mg/kg) and extract (200mg/kg and 400mg/kg) showed better protection against CCl4 induced toxicity to liver. Dunnet’s test showed a marked reduction in increased serum enzyme levels with extract treated animals compared to toxic control animals.

Table.1 Effect of ethanolic extract of Quisqualis indica on ALP, ALT, AST and SOD in Serum of Control and Experimental rats.

S.no TREATMENT ALP ALT AST SOD1 Normal 92.98±2.19 35.82±2.55 41.08±1.55 6.08±0.572 CCl4 induce 193.98±2.59*** 177.99±7.60*** 140.54±0.69*** 4.06±0.08**

(1ml/kg)

3 Silymarin 89.49±1.16ns 58.65±1.82** 40.48±1.11ns 6.70±0.21ns

(100mg/kg)4 Low Dose 114.16±2.42*** 80.81±1.25*** 56.35±1.38*** 4.93±0.41ns

(200 mg/kg)5 High Dose 100.54±2.65ns 69.86±1.66*** 47.84±0.68** 5.50±0.31ns

(400 mg/Kg)

Values are in mean±SE; Number of animals in each group = 4, ***p< 0.05 Group l Vs Group ll; *** ** *p<0/05 Vs Group ll. As compared with normal group (one way ANOVA followed by dunnet test)

250

200

150

ALP

ALT ALT

AST

SOD

100

50

Fig.2 Showing different Enzyme level between Normal, Induced and Treated.

PARACETAMOL-INDUCED HEPATOTOXICITY IN RATS

Result showed that on administration of paracetamol (750mg/kg; body weight), after 18 hours of intoxication resulted a significant (P<0.05) increase in hepatospecific serum biomarkers like ALT, AST, ALP, and decrease in SOD level in paracetamol-treated group, in comparison with the normal control group. On administration of Extract and Silymarin at the dose of 100mg/kg the level of these enzymes were found retrieving towards normalcy.The effects of ethanolic extract of Quisqualis indica on ALT, AST, ALP and SOD levels in paracetamol-induced liver damage in rats are summarized in Table2.

Table.2 Effect of ethanolic extract of Quisqualis indica on ALP, ALT, AST and SOD in Serum of Control and Experimental rats

S.no TREATMENT ALP ALT AST SOD1 Normal 92.89±1.55 37.37±1.96 40.76±1.26 5.80±0.392 Paracetamol induce 195.16±2.12*** 184.48±5.68*** 138.83±0.56*** 3.66±0.28**

(750mg/kg)

3 Silymarin 89.69±0.84ns 58.38±1.05*** 40.23±0.92ns 6.04±0.36ns

(100mg/kg)4 Low Dose 115.16±1.10*** 80.23±0.67*** 56.11±0.79*** 5.23±0.44ns

(200 mg/kg)5 High Dose 100.26±1.39** 69.29±1.08*** 47.84±0.62*** 6.42±0.29ns

(400 mg/Kg)

Values are in mean±SE; Number of animals in each group = 4, ***p< 0.05 Group l Vs Group ll; *** ** *p<0/05 Vs Group ll. As compared with normal group (one way ANOVA followed by dunnet test)

250

200

150

100

ALP

ALT

AST

SO D

50

0

Fig.3 Showing different Enzyme level between Normal, Induced and Treated.

DOXORUBICIN INDUCED MYOCARDIAL INFRACTION

On chronic administration of doxorubicin induced cardiac toxicity and effect of Quisqualis indica was established by significant elevated in cardiac biomarker enzymes. Animals treated with doxorubicin produced significant elevated in the levels of cholesterol, triglycerides compared to group 1 and there was very slight difference in HDL levels compared to group 1. Group 3 and 4 produced significant decrease in the level of cholesterol, triglycerides and increase in level of HDL as compared to group 2.Animals treated with doxorubicin produced significant increase in the levels of CPK and LDH compared to group 1, Group 3 and 4.

Table.3 Effect of ethanolic extract of Quisqualis indica on TC, TG, HDL, LDH, LDL and CK- MB in Serum of Control and Experimental rats

S.no TREATMENT TC TG HDL CK-MB LDH LDL1 Control 60.99±1.51*** 106.01±1.01*** 30.38±1.79* 19.99±0.67*** 149.14±2.91*** 28.92***2 Doxorubicin 107.06±2.21 297.81±1.53 22.15±1.03 34.61±1.33 205.04±2.31 72.47

induced(2.5mg/kg)

3 Low Dose 79.83±2.61*** 183.30±1.67*** 36.38±1.23ns 26.83±1.16*** 177.59±1.24*** 46.93***(200 mg/Kg)

4 High Dose 72.44±1.40** 159.69±1.11*** 35.9±3.41ns 21.97±0.54ns 165.34±1.82*** 38.42***(400mg/kg)

Values are in mean±SE; Number of animals in each group = 4, ***p< 0.05 Group l Vs Group ll; *** ** *p<0/05 Vs Group ll. As compared with control group (one way ANOVA followed by dunnet test)

400

300

200

100

Total cholestrol Triglyceride HDL

CK-MB LDH LDL

Fig.4-Showing different lipid serum level, cardiac biomarker between Normal, Induced

and Treated

HISTO PATHOLOGICAL OBSERVATIONS

Histology of liver sections of normal control animals (Group I normal contral) showed normal liver

architecture with were brought out central vein, were preserved cytoplasm and prominent nucleus

and nucleolus (Fig no: 5). The liver sections of CCl4 treated animals (Group II toxic control)

showed hepatic cells with serum toxicity characterized by inflammatory cell collection, scattered

inflammation across liver parenchyma, focal necrosis and swelling up of vascular endothelial cells

(Fig no:6).Silymarin (Group-V) exhibited protection from carbon tetrachloride induced changes in

the liver (Fig no.5). Ethanolic extract of Quisqualis indica pretreatment at a dose of 200mg and

400mg/kg (group III and IV) appeared to significantly prevent the CCl4 toxicity as revealed by the

hepatic cells with were preserved cytoplasm. Pretreatment also caused marked decrease in

inflammatory cells (Fig no:8 and 9).

HISTOPATHOLOGICAL STUDIES OFLIVER TISSUE BY CCL4 HEPATOTOXICITY

Fig.5. Liver section of group1 (Normal control)

Fig.no.6 Liver section of group2 (toxic control)

Fig.no.7 Liver section of Group3 (standard control)

Fig .no. 8 Liver section of Group 4 Extract 200 mg/kg/rat)

Fig.no. 9 Liver section of group 5 (Extract 400 mg/kg/rat)

DISCUSSION

Carbon tetrachloride has been well establishment as a liver toxicant, that induces the diffuse liver damage and its metabolites such as trichloromethyl radical (CCl3

•) and trichloromethyl peroxyl radical (CCl3O2

•) are also be noted that involved in the pathogenesis of liver and kidney damage. As we know that Liver and kidney are the two important vital organs mostly affected by the drugs.Liver damage is detected by the measurement of the levels of serum enzymes biomarkers like AST, ALT and ALP which has been released into the blood from damaged cells that means increased serum enzymes biomarkers are indicative of cellular leakage and loss of functional integrity of the hepatocyte and these are the important indicators of hepatic cell damage.When the plasma membrane of liver cells damaged, there are many different types of enzymes like as aspartate aminotransferase,alanine aminotransferase, alkaline phosphatase ,total bilurubin and gammaglutamyl transpeptidase are released into the blood stream. Their estimation in the serum is important significant as a quantitative marker of the extent and type of hepatocellular damage.When we founded that the normalization of the above serum biomarkers enzyme levels in rats treated with the extract clearly establishes the hepatoprotective effect of Quisqualis indica whichmight be able to induce accelerated regeneration of liver cells, reducing the leakage of the above enzymes into the blood. Superoxide dismutase (SOD), one of the important intracellular antioxidant enzymes, present in all aerobic cells has an antitoxic effect against superoxide anion.

In living systems, liver is a very vital organ that plays a vital role in metabolism and considered to be extremely sensitive to toxic agents. In the present study it was founded that the animals treated with acetaminophen resulted in significant hepatic damage as shown by the increased levels of serum markers. The pretreatment with extract, significantly attenuated the elevated levels of the serum markers. The normalization of serum biomarkers by extract suggested that they are capable to condition the hepatocytes so as to protect the membrane integrity against acetaminophen induced leakage of marker enzymes into the blood circulation. Acetaminophen induced leakage of marker enzymes into the blood circulation12-13.

Increase in serum level of ALP is due to increased synthesis in presence of increasing biliary pressure. Effective control of alkaline phosphatase activity points towards an early improvement in the secretory mechanism of the hepatic cell.In the present study, it was noted that the extract significantly elevated the hepatic SOD activity in paracetamol induced liver damage in rats. This show extract of Quisqualis indica can reduces reactive free radicals, that means ROS level also reduces that might lessen oxidative damage to the tissues and improve the activities of the hepatic antioxidant enzyme.The generation of ROS can be responsible for cellular damage leads to the cell death. In the present study, it was observed that extract of Quisqualis indica can reduces reactive free radicals that mean ROS level (superoxide radical, hydroxyl radicals and hydrogen peroxide), also reduces that leads to oxidative cellular damage hepatocyte also reduces.

The study entails the cadioprotective effect of Quisqualis indica against doxorubicin-induced cardiotoxicity. The present study is aimed to explore the cardioprotective effect of Quisqualis indica against doxorubicin induced cardiotoxicity in rats.

The existing experimental evidence indicates that doxorubicin oxidative stress is due to the production of free radicals in the heart tissue. The produced reactive oxygen species such as superoxide radicals, hydrogen peroxide and hydroxyl radicals are potential to cause damage to various intracellular components. The doxorubicin produced mitochondrial injury is critical to the heart because it would be highly adverse effects on the contractile functioning of the cardiac myocytes by alterations in the energy metabolism. On administration of Quisqualis indica was capable to reduce the doxorubicin-induced cardiotoxic manifestations in multiple ways. Elevated in the level of plasma triglycerides, total cholesterol and HDL in the doxorubicin induced group indicate doxorubicin may be interfering with metabolism or biosynthesis of lipids. In the present study it was observed that on pretreatment with Quisqualis indica also showed reduction in blood lipid profile levels with concomitantly increase in HDL cholesterol was observed. Reduction in the blood lipid profiles and increase in HDL cholesterol in Quisqualis indica treated group. Lipid lowering effect of Quisqualis indica is due to inhibition of hepatic cholesterol biosynthesis, increased fecal bile acid secretion and stimulation of receptor mediated catabolism of LDL. Heart tissue injury by doxorubicin in rats was showed by elevated level of the marker enzymes such as LDH and CK-MB. Quisqualis indica was found to inhibit the doxorubicin induced CPK and LDH release in the release in the serum of rats14.

CONCLUSION:

In conclusion our finding demonstrated that administration of ethanolic extract of 200 and 400 mg/kg of Quisqualis indica was effective to protect against oxidative stress and possess hepatoprotective antioxidant and cardioprotective. The active ingredients present here may recover the disorders in hepatic damage, and cardiotoxic state. Our result indicate that the extracts of Quisqualis indica produce a significant reduction of ALT AST and ALP and raised the SOD level which is showed more promising hepatoprotective and antioxidant activity Similarly it also inhibit the elevated markers enzyme (CPK, LDH), thus also act as cardioprotective drug and we also concluded that Quisqualis indica had lesser side effect than silymarin.

ACKNOWLEDGEMENTS

We are thankful to Director, Advance Institute of biotech and Paramedical Sciences, for their kindly support for my work. We are grateful to the technical staff members of the Department of Pharmacology. We also thank the following persons: , Sashi Pratap Singh, Yatendra Singh, Sagar Singh, Tanu Dixit Anuska Singh.

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