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1.  Hypoglycaemic and Hypolipidaemic Effects of Withania somnifera Root and Leaf Extracts on Alloxan-Induced Diabetic Rats 
Withania somnifera is an important medicinal plant, which is used in traditional medicine to cure many diseases. Flavonoids were determined in the extracts of W. somnifera root (WSREt) and leaf (WSLEt). The amounts of total flavonoids found in WSREt and WSLEt were 530 and 520 mg/100 g dry weight (DW), respectively. Hypoglycaemic and hypolipidaemic effects of WSREt and WSLEt were also investigated in alloxan-induced diabetic rats. WSREt and WSLEt and the standard drug glibenclamide were orally administered daily to diabetic rats for eight weeks. After the treatment period, urine sugar, blood glucose, haemoglobin (Hb), glycosylated haemoglobin (HbA1C), liver glycogen, serum and tissues lipids, serum and tissues proteins, liver glucose-6-phosphatase (G6P) and serum enzymes like aspartate transaminase (AST), alanine transaminase (ALT), acid phosphatase (ACP) and alkaline phosphatase (ALP) levels were determined. The levels of urine sugar, blood glucose, HbA1C, G6P, AST, ALT, ACP, ALP, serum lipids except high density lipoprotein-bound cholesterol (HDL-c) and tissues like liver, kidney and heart lipids were significantly (p < 0.05) increased, however Hb, total protein, albumin, albumin:globulin (A:G) ratio, tissues protein and glycogen were significantly (p < 0.05) decreased in alloxan-induced diabetic rats. Treatment of the diabetic rats with WSREt, WSLEt and glibenclamide restored the changes of the above parameters to their normal level after eight weeks of treatment, indicating that WSREt and WSLEt possess hypoglycaemic and hypolipidaemic activities in alloxan-induced diabetes mellitus (DM) rats.
doi:10.3390/ijms10052367
PMCID: PMC2695282  PMID: 19564954
alloxan; diabetic; Withania somnifera; hypoglycaemic; hypolipidaemic; flavonoids
2.  Cardioprotective effect of Saraca indica against cyclophosphamide induced cardiotoxicity in rats: A biochemical, electrocardiographic and histopathological study 
Indian Journal of Pharmacology  2013;45(1):44-48.
Objectives:
Cardioprotective activity of alcoholic extract of Saraca indica (SI) bark was investigated against cyclophosphamide induced cardiotoxicity.
Materials and Methods:
Cardiotoxicity was induced in Wistar rats by administering cyclophosphamide (200 mg/kg, i.p.) single injection on first day of experimental period. Saraca indica (200 and 400 mg/kg, p.o.) was administered immediately after administration of cyclophosphamide on first day and daily for 10 days. The general observations and mortality were measured.
Results:
Cyclophosphamide administration significantly (p < 0.05) increased lipid peroxidation (LPO) and decreased the levels of antioxidant markers such as reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT). Cyclophosphamide elevated the levels of biomarker enzymes like creatine kinase (CK), creatine kinase isoenzyme MB (CK-MB), lactate dehydrogenase (LDH), aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP). Further, the cyclophosphamide treated rats showed changes in electrocardiogram (ECG) along with increased levels of cholesterol and triglycerides. Treatment with Saraca indica significantly (p < 0.05) reversed the status of cardiac biomarkers, ECG, oxidative enzymes and lipid profile in cyclophosphamide induced cardiotoxicity. Potential cardioprotective effect of Saraca indica was supported by histopathological examination that reduced severity of cellular damage of the myocardium.
Conclusion:
The biochemical, ECG and histopathology reports support the cardioprotective effect of Saraca indica which could be attributed to antioxidant activity.
doi:10.4103/0253-7613.106434
PMCID: PMC3608294  PMID: 23543849
Cardioprotective; cyclophosphamide; ECG; free radicals; Saraca indica Linn
3.  Antiproliferative role of Indigofera aspalathoides on 20 methylcholanthrene induced fibrosarcoma in rats 
Objective
To find out the anticancer effect of Indigofera aspalathoides (I. aspalathoides) on 20-methylcholanthrene induced fibrosarcoma in rats.
Methods
Fibrosarcoma was induced in Wistar strain male albino rats by 20-methylcholanthrene. Intraperitoneous (i.p.) administration of 250 mg/kg body weight/day of aqueous extract of I. aspalathoides for 30 d effectively suppressed chemically induced tumors. Parameters such as body weight, liver and kidney weight, tumor weight, mean survival time, behavioral changes, blood glucose, blood glycogen and marker enzymes such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), acid phosphatase (ACP) and 5′-nucleiotidase (5′-NT) in serum, liver and kidney and lipid profiles such as total cholesterol, phospholipids, free fatty acids in liver and kidney of control and experimental animals were studied.
Results
Fibrosarcoma bearing animals were ferocious and anxious. The mean survival time was found to increase after the treatment. The body weights were significantly decreased (P<0.001) in group II fibrosarcoma animals which steadily increased after the treatment with I. aspalathoides. The liver and kidney weights were significantly increased whereas the tumor weights decreased as compared to the weights in untreated fibrosarcoma bearing rats. The blood glucose and the liver and kidney glycogen levels were found to decrease significantly (P<0.001) in group II animals. Elevated activities of marker enzymes were observed in serum, liver and kidney of fibrosarcoma bearing Group II animals which were normalize after I. aspalathoides treatment. In the liver and kidney of Group II animals the total cholesterol increased whereas the phospholipids and free fatty acid levels decreased (P<0.001) which were normalized after treatment.
Conclusions
The treatment by I. aspalathoides on fibrosarcoma bearing rats has improved the levels of various parameters indicating its antiproliferative and anticancer activity.
doi:10.1016/S2221-1691(13)60008-8
PMCID: PMC3621473  PMID: 23593577
Chemoprevention; Tumor weight; Mean survival time; Glucose; Glycogen; Marker enzymes
4.  EFFECT OF ETHANOLIC FRUIT EXTRACT OF Pedalium murex Linn. IN ETHYLENE GLYCOL INDUCED UROLITHIASIS IN MALE WISTAR ALBINO RATS 
Ancient Science of Life  2010;29(4):29-34.
The ethanolic fruit extract of Pedalium murex to ethylene glycol intoxicated rats reverted the levels of the liver and kidney markers to near normal levels protecting liver and renal tissues from damage and also prevents the crystal retention in tissues. The levels of ACP, ALP, AST, ALT in serum andurine were significantly increased due to the damaged structural integrity of renal and hepatic cells causing the enzymes which are located in the cytoplasm to be released into the circulation. The levels of ACP and ALP, AST, ALT in renal and hepatic tissues of ethylene glycol induced rats might be due to leakage of the enzyme into the general circulation from the collateral circulation. LDH levels in serum, urine and tissues were increased on ethylene glycol intoxication is due to the oxalate induced renal and hepatic cellular damage.
PMCID: PMC3336290  PMID: 22557365
Marker enzymes; urolithiasis; ethylene glycol; Pedalium murex
5.  Antioxidant and hepatoprotective effect of Garcinia indica fruit rind in ethanol-induced hepatic damage in rodents 
Interdisciplinary Toxicology  2012;5(4):207-213.
The protective effects of aqueous extracts of the fruit rind of Garcinia indica (GIE) on ethanol-induced hepatotoxicity and the probable mechanisms involved in this protection were investigated in rats. Liver damage was induced in rats by administering ethanol (5 g/kg, 20% w/v p.o.) once daily for 21 days. GIE at 400 mg/kg and 800 mg/kg and the reference drug silymarin (200 mg/kg) were administered orally for 28 days to ethanol treated rats, this treatment beginning 7 days prior to the commencement of ethanol administration. Levels of marker enzymes (aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP)), triglyceride (sTG), albumin (Alb) and total protein (TP) were evaluated in serum. Antioxidant parameters (reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR)), hepatic triglycerides (hTG) and the lipid peroxidation marker malondialdehyde (MDA) were determined in liver. GIE and silymarin elicited significant hepatoprotective activity by attenuating the ethanol–elevated levels of AST, ALT, ALP, sTG, hTG and MDA and restored the ethanol-depleted levels of GSH, SOD, CAT, GPx, GR, Alb and TP. GIE 800 mg/kg demonstrated greater hepatoprotection than GIE 400 mg/kg. The present findings indicate that hepatoprotective effects of GIE in ethanol-induced oxidative damage may be due to an augmentation of the endogenous antioxidants and inhibition of lipid peroxidation in liver.
doi:10.2478/v10102-012-0034-1
PMCID: PMC3600525  PMID: 23554565
Garcinia indica; ethanol; hepatoprotective; antioxidant activity
6.  Evaluation of Hepatoprotective Effect of Leaves of Cassia sophera Linn. 
In the present study, the hepatoprotective activity of ethanolic extracts of Cassia sophera Linn. leaves was evaluated against carbon-tetrachloride- (CCl4-) induced hepatic damage in rats. The extracts at doses of 200 and 400 mg/kg were administered orally once daily. The hepatoprotection was assessed in terms of reduction in histological damage, changes in serum enzymes, serum glutamate oxaloacetate transaminase (AST), serum glutamate pyruvate transaminase (ALT), serum alkaline phosphatase (ALP), total bilirubin, and total protein levels. The substantially elevated serum enzymatic levels of AST, ALT, ALP, and total bilirubin were restored towards the normalization significantly by the extracts. The decreased serum total protein level was significantly normalized. Silymarin was used as standard reference and exhibited significant hepatoprotective activity against carbon tetrachloride-induced hepatotoxicity in rats. The biochemical observations were supplemented with histopathological examination of rat liver sections. The results of this study strongly indicate that Cassia sophera leaves have potent hepatoprotective action against carbon tetrachloride-induced hepatic damage in rats. This study suggests that possible activity may be due to the presence of flavonoids in the extracts.
doi:10.1155/2012/436139
PMCID: PMC3368335  PMID: 22690244
7.  Effect of vitamin E alone and in combination with lycopene on biochemical and histopathological alterations in isoproterenol-induced myocardial infarction in rats 
Background:
The present study has been designed to evaluate the combined cardioprotective effect of vitamin E and lycopene on biochemical and histopathological alteration in isoproterenol-induced myocardial infarction in rats.
Materials and Methods:
Adult male albino rats of Wistar strain were treated with isoproterenol (200 mg/kg, s.c.) for 2 days at an interval of 24 h to develop myocardial infarction. Vitamin E (100 mg/kg/day, p.o.) and lycopene (10 mg/kg/day, p.o.) were administered alone and in combination for 30 days. Change in body weight and organ weight were monitored. Levels of serum marker enzymes (AST, ALT, LDH and CK-MB), lipid peroxidation, endogenous antioxidants (GSH, GPX, GST, SOD and CAT), membrane bound enzymes (Na+/K+ ATPases, Mg2+ ATPases and Ca2+ ATPases) were evaluated. LDH isoenzyme separation was carried out using gel electrophoresis. Histopathology of heart tissue was performed.
Results:
Induction of rats with isoproterenol resulted in a significant elevation in organ weight, lipid peroxidation, serum marker enzymes (AST, ALT, CK-MB and LDH), and Ca 2+ ATPases, whereas it caused a significant (P < 0.001) decrease in body weight, activities of endogenous antioxidants (GSH, GPx, GST, SOD and CAT), Na+/K+ and Mg2+ ATPases. ISO treated rats showed high intensity band of LDH1-LDH2 isoenzymes. Treatment with the combination of Vitamin E and lycopene for 30 days significantly attenuated these changes as compared to the individual treatment and ISO treated groups. Histopathological observations were also in correlation with the biochemical parameters.
Conclusion:
These findings indicate the synergistic cardioprotective effects of vitamin E and lycopene during ISO-induced myocardial infarction in rats.
doi:10.4103/0976-500X.64532
PMCID: PMC3142754  PMID: 21808587
Isoproterenol; myocardial infarction; oxidative stress; vitamin E; lycopene
8.  Salivary enzymes as diagnostic markers for detection of gingival/periodontal disease and their correlation with the severity of the disease 
Context:
Host responses to periodontal disease include the production of different enzymes released by stromal, epithelial or inflammatory cells. Important enzymes associated with cell injury and cell death are aspartate aminotransferase, alanine aminotransferase (AST, ALT), alkaline phosphatase, acidic phosphatase (ALP, ACP), and gama glutamyl transferase (GGT). Changes in enzymatic activity reflect metabolic changes in the gingiva and periodontium, in the inflammation.
Aims:
In this article we examined the activity of AST, ALT, GGT, ALP, and ACP in the saliva from patients with periodontal disease, before and after periodontal treatment (experimental group — 20 gingivitis patients and 20 periodontitis patients), and in the saliva from healthy subjects (control group — 20 samples).
Settings and Design:
Periodontal disease was determined based on the clinical parameters (gingival index (GI), probing depth (PD), and clinical attachment loss (CAL)). Patients with periodontal disease were under conventional periodontal treatment.
Materials and Methods:
The stimulated saliva of the patient was collected in a sterile test tube and analyzed using the Automatic Analyzer.
Results:
The obtained results showed statistically significant increased activity of AST, ALT, GGT, ALP, and ACP in the saliva from patients with periodontal disease, in relation to the control group. A significant reduction in the enzyme levels was seen after conventional periodontal therapy.
Conclusions:
Based on these results, it can be assumed that the salivary enzymes (AST, ALT, GGT, ALP, and ACP) can be considered as biochemical markers for evaluating the diagnosis and prognosis of the functional condition of periodontal tissues in disease and health, and in the evaluation of the therapy effects in periodontal disease.
doi:10.4103/0972-124X.100911
PMCID: PMC3498704  PMID: 23162329
Enzymes; periodontal disease; saliva
9.  The gold nanoparticle size and exposure duration effect on the liver and kidney function of rats: In vivo 
Nanoparticles (NPs) offer a great possibility for biomedical application, not only to deliver pharmaceutics, but also to be used as novel diagnostic and therapeutic approaches. Currently, there are no data available regarding to what extent the degree of the toxicity and the accumulation of gold nanoparticles (GNPs) are present in in vivo administration. This study aimed to address the GNP size and exposure duration effect on the liver and kidney function of rats: in vivo.
Methods
A total of 30 healthy male Wistar-Kyoto rats of the same age (12 weeks old) and weighing 220–240 g of King Saud University colony were used. Animals were randomly divided into groups, two GNP-treated rat groups and one control group (CG). The 50 μl of 10 and 50 nm GNPs was intraperitoneally administered in rats for exposure duration of 3 days. Then, several biochemical parameters such as aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alanine transaminase (ALT), alkaline phosphatase (ALP), urea (UREA) and creatinine (CREA) were evaluated.
Results
In this study, the AST values increased with the administration of 10 and 50 nm GNPs compared with the control. The AST values significantly increased with 10 nm GNPs compared with 50 nm GNPs and control. The GGT and ALT values decreased with the administration of 10 and 50 nm GNPs compared with the control. The GGT and ALT values significantly decreased with 50 nm GNPs compared with 10 nm GNPs and control. The ALP values significantly decreased with the administration of 10 and 50 nm GNPs compared with the control. The decrease in ALP values with 10 nm GNPs was higher than those compared with 50 nm GNPs. In this study, the levels of UREA and CREA values increased in a non significant manner after the administration of 10 and 50 nm GNPs compared with the control.
Conclusions
This study demonstrates that the increase in the enzymes AST and the decrease in ALP are smaller GNPs (10 nm) size-dependent for exposure duration of 3 days; while the decrease in the enzymes GGT and ALT are bigger GNPs (50 nm) size-dependent. The levels of UREA and CREA values indicated no significant changes with the administration of 10 and 50 nm GNPs for exposure duration of 3 days compared with the control. The administration of 10 and 50 nm GNPs for short exposure duration of 3 days induced only significant variations with some liver enzymes while kidney showed no significant variations. This study suggests that synthesis and metabolism of GNPs as well as the protection of the liver will be more important issues for medical applications of gold-based nanomaterials in future.
doi:10.1016/j.sjbs.2013.01.007
PMCID: PMC3730801  PMID: 23961234
Gold nanoparticle; Sizes; Exposure duration; Rats; Liver and kidney function
10.  Prevention of hepatorenal toxicity with Sonchus asper in gentamicin treated rats 
Background
Sonchus asper possesses antioxidant capacity and is used in liver and kidney disorders. We have investigated the preventive effect of methanolic extract of Sonchus asper (SAME) on the gentamicin induced alterations in biochemical and morphological parameters in liver and kidneys of Sprague-Dawley male rat.
Methods
Acute oral toxicity studies were performed for selecting the therapeutic dose of SAME. 30 Sprague-Dawley male rats were equally divided into five groups with 06 animals in each. Group I received saline (0.5 ml/kg bw; 0.9% NaCl) while Group II administered with gentamicin 0.5 ml (100 mg/kg bw; i.p.) for ten days. Animals of Group III and Group IV received gentamicin and SAME 0.5 ml at a dose of 100 mg/kg bw and 200 mg/kg bw, respectively while Group V received only SAME at a dose of 200 mg/kg bw. Biochemical parameters including aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), γ-glutamyltransferase (γ-GT), total cholesterol, triglycerides, total protein, albumin, creatinine, blood urea nitrogen (BUN), total bilirubin and direct bilirubin were determined in serum collected from various groups. Urinary out puts were measured in each group and also assessed for the level of protein and glucose. Lipid peroxides (TBARS), glutathione (GSH), DNA injuries and activities of antioxidant enzymes; catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) were determined in liver and renal samples. Histopathological studies of liver and kidneys were also carried out.
Results
On the basis of acute oral toxicity studies, 2000 mg/kg bw did not induce any toxicity in rats, 1/10th of the dose was selected for preventive treatment. Gentamicin increased the level of serum biomarkers; AST, ALT, ALP, LDH, γ-GT, total cholesterol, triglycerides, total protein, albumin, creatinine, BUN, total and direct bilirubin; as were the urinary level of protein, glucose, and urinary output. Lipid peroxidation (TBARS) and DNA injuries increased while GSH contents and activities of antioxidant enzymes; CAT, POD, SOD decreased with gentamicin in liver and kidney samples. SAME administration, dose dependently, prevented the alteration in biochemical parameters and were supported by low level of tubular and glomerular injuries induced with gentamicin.
Conclusion
These results suggested the preventive role of SAME for gentamicin induced toxicity that could be attributed by phytochemicals having antioxidant and free radical scavenging properties.
doi:10.1186/1472-6882-11-113
PMCID: PMC3305921  PMID: 22082144
11.  EFFECT OF TERMINALIA ARJUNA STEM BARK EXTRACT ON THE ACTIVITIES OF MARKER ENZYMES IN ALLOXAN INDUCED DIABETIC RATS 
Ancient Science of Life  2005;25(1):8-15.
Insight of evidence that some complications of diabetes mellitus due to hyperglycemia, we investigated the effect of T. arjuna bark extract on serum, liver and kidney marker enzymes in alloxan - induced diabetic rats. T. arjuna was administered orally at a doses of 250 and 500 mg/kg body weight for 30 days, after which serum liver and kidney tissues were assayed for the degree of pathological changes by means of markers such as alkaline phosphatase (ALP), acid phosphatase (ACP), alanine amino transferase (ALT), aspartate amino transferase (AST) and lactate dehydrogenase (LDH) resulted in a significant reduction in serum and tissue of liver and kidney marker enzymes when compared with control rats T. arjuna at a dose of 500 mg/kg body weight exhibited higher efficacy.
PMCID: PMC3330897  PMID: 22557182
Terminalia arjuna; hyperglycemia; alloxan diabetes; marker enzymes
12.  Effects of Echinacea purpurea on Hepatic and Renal Toxicity Induced by Diethylnitrosamine in Rats 
Background
Nitrites are mainly used in food preservation. These materials could change to nitrosamine due to the effect of heat and gastric acid. Nitrosamine is absorbed in intestine and enters the liver and hepatocytes by portal venous system, and hampers the detoxification system of liver by interfering in cytochrome P450 enzymes, so, the liver gently proceeds to cirrhosis and cancer.
Objectives
The current study aimed to investigate the hepatic and renal protective effects of aerial parts of Echinacea purpurea extract (EPE) on injury induced by diethylnitrosamine (DEN).
Materials and Methods
Twenty Wistar rats were divided into 4 groups. Groups were as follow: Control group (normal saline), DEN (200 mg/kg, IP, a single dose), EPE (100 mg/kg, orally, daily) and DEN + EPE which received as group DEN and EPE. After 30 days, Blood samples, and liver and kidney tissues were taken for further examination. Aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), BUN, Creatinine and total and direct bilirubin were estimated in serum.
Results
DEN induced hepatotoxicity and nephrotoxicity in all the treated animals by elevated serum ALT, AST, ALP and BUN, creatinin and total and direct bilirubin levels. AST, BUN and total and direct bilirubin significantly decreased in DEN + EPE compared to DEN group. After 30 days of DEN administration, histopathological investigation revealed proliferation of hepatic stellate cells and early fibrosis which were partly improved by EPE administration.
Conclusions
The current study findings indicated that Echinacea purpurea extract played an important role in the protection against DEN toxicity in rats.
PMCID: PMC3941908  PMID: 24624189
Diethylnitrosamine; Echinacea; Rats; Liver; Kidney; Toxicity
13.  Chemoprotective potential of Coccinia indica against cyclophosphamide-induced toxicity 
Indian Journal of Pharmacology  2013;45(5):502-507.
Objective:
Although cyclophosphamide (CP), an alkylating agent, is used in the treatment of cancer owing to its broad-spectrum efficacy, its metabolites exhibit severe undesired toxicities in normal cells. The present study was aimed to investigate the chemoprotective potential of Coccinia indica against CP-induced oxidative stress, genotoxicity, and hepatotoxicity.
Materials and Methods:
Rodents were orally pre-treated with Coccinia indica extract (200, 400, and 600 mg/kg) for five consecutive days. On 5th day, these animals were injected with CP (50 mg/kg i.p) and sacrificed after 24 hrs. for the evaluation of oxidative stress, hepatotoxicity, micronucleus formation, and chromosomal aberrations.
Results:
We found that the CP significantly increased malondialdehyde (MDA) and decreased catalase and glutathione (GSH) levels in brain, and it was significantly reversed by Coccinia indica extract (400 and 600 mg/kg). Further, pre-treatment with Coccinia indica extract (200, 400, 600 mg/kg) significantly and dose-dependently reduced micronuclei formation and incidence of aberrant cells. We also found that the CP-induced increase in the serum biomarker enzymes like alkaline phosphatase (ALP), alkaline aminotransferase (ALT), and aspartate aminotransferase (AST) were significantly reduced by Coccinia indica extract.
Conclusion:
Thus, the present results indicate the protective effect of Coccinia indica extract against CP-induced oxidative stress, genotoxicity, as well as hepatotoxicity.
doi:10.4103/0253-7613.117783
PMCID: PMC3793523  PMID: 24130387
Coccinia indica; cyclophosphamide; genotoxicity; oxidative stress
14.  Hepatoprotective effects of methanol extract of Carissa opaca leaves on CCl4-induced damage in rat 
Background
Carissa opaca (Apocynaceae) leaves possess antioxidant activity and hepatoprotective effects, and so may provide a possible therapeutic alternative in hepatic disorders. The effect produced by methanolic extract of Carissa opaca leaves (MCL) was investigated on CCl4-induced liver damages in rat.
Methods
30 rats were divided into five groups of six animals of each, having free access to food and water ad libitum. Group I (control) was given olive oil and DMSO, while group II, III and IV were injected intraperitoneally with CCl4 (0.5 ml/kg) as a 20% (v/v) solution in olive oil twice a week for 8 weeks. Animals of group II received only CCl4. Rats of group III were given MCL intragastrically at a dose of 200 mg/kg bw while that of group IV received silymarin at a dose of 50 mg/kg bw twice a week for 8 weeks. However, animals of group V received MCL only at a dose of 200 mg/kg bw twice a week for 8 weeks. The activities of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and γ-glutamyltransferase (γ-GT) were determined in serum. Catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), glutathione-S-transferase (GST), glutathione peroxidase (GSH-Px), glutathione reductase (GSR) and quinone reductase (QR) activity was measured in liver homogenates. Lipid peroxidation (thiobarbituric acid reactive substances; TBARS), glutathione (GSH) and hydrogen peroxide (H2O2) concentration was also assessed in liver homogenates. Phytochemicals in MCL were determined through qualitative and high performance liquid chromatography (HPLC) analysis.
Results
Hepatotoxicity induced with CCl4 was evidenced by significant increase in lipid peroxidation (TBARS) and H2O2 level, serum activities of AST, ALT, ALP, LDH and γ-GT. Level of GSH determined in liver was significantly reduced, as were the activities of antioxidant enzymes; CAT, POD, SOD, GSH-Px, GSR, GST and QR. On cirrhotic animals treated with CCl4, histological studies showed centrilobular necrosis and infiltration of lymphocytes. MCL (200 mg/kg bw) and silymarin (50 mg/kg bw) co-treatment prevented all the changes observed with CCl4-treated rats. The phytochemical analysis of MCL indicated the presence of flavonoids, tannins, alkaloids, phlobatannins, terpenoids, coumarins, anthraquinones, and cardiac glycosides. Isoquercetin, hyperoside, vitexin, myricetin and kaempherol was determined in MCL.
Conclusion
These results indicate that MCL has a significant protective effect against CCl4 induced hepatotoxicity in rat, which may be due to its antioxidant and membrane stabilizing properties.
doi:10.1186/1472-6882-11-48
PMCID: PMC3141600  PMID: 21699742
Carissa opaca; Carbon tetrachloride; Hepatotoxicity; Oxidative stress; Phytochemical analysis
15.  Protective Effects of Garlic and Silymarin on NDEA-Induced Rats Hepatotoxicity 
Background ­— The present study was conducted to investigate the chemopreventive effects of garlic extract and silymarin on N-nitrosodiethylamine (NDEA) and carbon tetrachloride (CCl4)-induced hepatotoxicity in male albino rats. Methods and Results — Animals were pretreated with garlic, silymarin or both for one week prior to the injection of NDEA. Then animals received a single injection of NDEA followed by weekly subcutaneous injections of CCl4 for 6 weeks. Oral administration was then continued along with the injection of CCl4 for the duration of the experiment. Serum aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), hepatic lipid peroxidation (LPO), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione-S-transferase (GST) and glutathione reductase (GSR) were measured. Injection of NDEA induced a significant elevation in serum AST, ALT and ALP. In the liver, NDEA increased oxidative stress through the increase in LPO and decrease in SOD, and GSH-dependent enzymes. Although administration of garlic or silymarin significantly reduced the liver toxicity, combined administration was more effective in preventing the development of hepatotoxicity. Conclusion — These novel findings suggest that silymarin and garlic have a synergistic effect, and could be used as hepatoprotective agents against hepatotoxicity.
PMCID: PMC2737715  PMID: 19742242
Hepatotoxicity; NDEA; Garlic; Silymarin; liver enzymes; oxidative stress; rats.
16.  Protective Role of Tinospora cordifolia against Lead-induced Hepatotoxicity 
Toxicology International  2010;17(1):12-17.
The importance of Tinospora cordifolia stem and leaves extract was investigated for its possible hepatoprotective effect in Swiss albino male mice against lead nitrate induced toxicity. Oral administration of plant extracts prevented the occurrence of lead nitrate induced liver damage. The decreased level of tissue enzymes, i.e., superoxide dismutase (SOD), catalase (CAT) and increased level of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and acid phosphatase (ACP) were observed in mice treated with lead. Administration of aqueous stem extract (400 mg/kg body weight, orally) and aqueous leaves extract (400 mg/kg body weight, orally) along with the lead nitrate (5 mg/kg body weight, i.p. for 30 days) increased the activities of SOD and CAT and decreased the levels of AST, ALT, ALP, and ACP enzymes in mice. These biochemical observations were supplemented by histopathology/histological examinations of liver section. Results of this study revealed that plant extract could afford protection against lead-induced hepatic damage.
doi:10.4103/0971-6580.68343
PMCID: PMC2964743  PMID: 21042467
Biochemical changes; histopathology; lead nitrate; liver; mice; Tinospora cordifolia
17.  Hepatoprotective effect of Pterocarpus marsupium against carbon tetrachloride induced damage in albino rats 
Ancient Science of Life  2007;27(1):19-25.
Medicinal plants play a key role in human health care. Pterocarpus marsupium is one of the plants used in treatment of diabetes mellitus and the present study was aimed to assess hepatoprotective effect of the plant against CCl4 induced hepatotoxicity. Wistar albino rats were divided into four groups. Group I was normal control group; Group II, the hepatotoxic group was given CCl4 (2ml/kg body weight intraperitoneally); Groups III received CC14 + Plant extract (100 mg/kg b.w orally); Group IV received only the plant extract. Liver markers were assayed in serum and liver tissue. Levels of marker enzymes such as alanine transminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) and bilirubin were increased significantly in Group II. These enzymes were significantly decreased in Group III treated with plant extracts. The present investigation suggest that the plant had a good protective effect on CCl4 induced hepatic injury.
PMCID: PMC3330841  PMID: 22557255
18.  Eleusine indica L. possesses antioxidant activity and precludes carbon tetrachloride (CCl4)-mediated oxidative hepatic damage in rats 
Objectives
The purpose of this study was to evaluate the ability of aqueous extract of Eleusineindica to protect against carbon tetrachloride (CCl4)-induced hepatic injury in rats.
Methods
The antioxidant activity of E. indica was evaluated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay. The total phenolic content of E. indica was also determined. Biochemical parameters [e.g. alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), glutathione (GSH), catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase and quinone reductase] were used to evaluate hepatic damage in animals pretreated with E. indica and intoxicated with CCl4. CCl4-mediated hepatic damage was also evaluated by histopathologically.
Results
E. indica extract was able to reduce the stable DPPH level in a dose-dependent manner. The half maximal inhibitory concentration (IC50) value was 2350 μg/ml. Total phenolic content was found to be 14.9 ± 0.002 mg/g total phenolic expressed as gallic acid equivalent per gram of extract. Groups pretreated with E. indica showed significantly increased activity of antioxidant enzymes compared to the CCl4-intoxicated group (p < 0.05). The increased levels of serum ALT and AST were significantly prevented by E. indica pretreatment (p < 0.05). The extent of MDA formation due to lipid peroxidation was significantly reduced (p < 0.05), and reduced GSH was significantly increased in a dose-dependently manner (p < 0.05) in the E. indica-pretreated groups as compared to the CCl4-intoxicated group. The protective effect of E. indica was further evident through decreased histopathological alterations in the liver.
Conclusion
The results of our study indicate that the hepatoprotective effects of E. indica might be ascribable to its antioxidant and free radical scavenging property.
doi:10.1007/s12199-011-0255-5
PMCID: PMC3390565  PMID: 22207570
E. indica; Antioxidant activity; Hepatoprotective effects; Oxidative stress; Carbon tetrachloride
19.  Pioglitazone, quercetin and hydroxy citric acid effect on hepatic biomarkers in Non Alcoholic Steatohepatitis 
Pharmacognosy Research  2014;6(2):153-162.
Background:
Non alcoholic steatohepatitis (NASH), severe form of diseases belonging to the spectrum of the Non alcoholic fatty liver disease (NAFLD). It is an asymptomatic disease which leads to fibrosis and finally to cirrhosis, an end stage liver disease.
Objective:
To study the effect of pioglitazone, quercetin and hydroxy citric acid on hepatic biomarkers and various biochemical parameters in experimentally induced non alcoholic steatohepatitis (NASH).
Materials and Methods:
Male Wister rats were divided into 8 groups. The activities of alkaline phosphatase (ALP), aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH) and γ-Glutamyl Transferase (GGT) were assayed in serum. The levels of various other biochemical parameters such as serum albumin, total bilirubin, creatinine, urea, uric acid and glucose were also estimated in experimental NASH.
Results:
The NASH group produced severe liver injury by significantly increasing the serum levels of ALT, AST, GGT and LDH compared with that of the control. However, the experimental NASH rats treated with pioglitazone, with quercetin and with hydroxy citric acid showed an obvious decrease in ALT, AST, GGT and LDH levels when compared with that of NASH induced group. A significant increase in the levels of albumin, creatinine, urea, uric acid, glucose and total bilirubin was noticed in experimentally induced NASH group (group 2) when compared to rats in control group (group 1).
Conclusion:
It could be inferred from this study that, pioglitazone, quercetin and hydroxy citric acid may afford protection to the liver against NASH, as evidenced by the results of this study on the levels of various biochemical parameters such as glucose, urea, uric acid, creatinine and bilirubin. Whereas from the results of hepatic marker enzymes, it is evident that optimal protection was observed after quercetin treatment against experimental NASH whereas pioglitazone and hydroxy citric acid also confers protection to some extent against NASH.
doi:10.4103/0974-8490.129037
PMCID: PMC3996753  PMID: 24761121
Biochemical parameters; experimentally induced NASH; hydroxy citric acid; liver marker enzymes; non alcoholic steatohepatitis; pioglitazone; non alcoholic fatty liver disease; non alcoholic steatohepatitis; quercetin
20.  Protective Effect of Quercetin on Atrazine-Induced Oxidative Stress in the Liver, Kidney, Brain, and Heart of Adult Wistar Rats 
Toxicology International  2014;21(2):148-155.
Background:
The conflicting roles of quercetin against tissue pathologies associated with oxidative stress are known.
Objective:
To evaluate the effect of quercetin at doses of 5 mg/kg (Q5) or 10 mg/kg (Q10) against atrazine (120 mg/kg, ATZ)-induced oxidative stress in various tissues of rats.
Materials and Methods:
Adult male albino Wistar rats were administered ATZ, Q5, and Q10 alone or in combination for 16 days. At the end of the 16th day, the animals were sacrificed by cervical dislocation; and the blood, heart, brain, kidney and liver were collected and used for biochemical determinations and histopathological examination.
Results:
Q10 but not Q5 attenuated ATZ-induced increase in the levels of serum enzyme markers sorbitol dehydrogenase (SDH), acid phosphatase (ACP), alkaline phosphatase (ALP), and aspartate aminotransferase (AST). The heart was less susceptible to ATZ-induced oxidative stress than the liver, kidney, and brain of treated animals, and there were tendencies for synergistic effects in the heart and liver of Q5 + ATZ-treated rats. Oxidative stress-induced by ATZ in terms of increased lipid peroxidation level and superoxide dismutase (SOD) activity was decreased in the brain of the Q5 + ATZ-treated rats but not that of the Q10 + ATZ-treated rats. Conversely, histopathological changes and oxidative stress-induced by ATZ in terms of elevated lipid peroxidation level, decreased SOD, and catalase (CAT) activities were prevented in the kidney and liver of the Q10 + ATZ-treated rats but not that of the Q5 + ATZ-treated rats.
Conclusion:
Quercetin at the investigated doses and especially the low dose may not protect against ATZ-induced oxidative stress in rat tissues in an overall sense.
doi:10.4103/0971-6580.139794
PMCID: PMC4170555  PMID: 25253923
Atrazine; malondialdehyde; oxidative stress; quercetin; serum marker enzymes
21.  Dose-response relationship between arsenic exposure and the serum enzymes for liver function tests in the individuals exposed to arsenic: a cross sectional study in Bangladesh 
Environmental Health  2011;10:64.
Background
Chronic arsenic exposure has been shown to cause liver damage. However, serum hepatic enzyme activity as recognized on liver function tests (LFTs) showing a dose-response relationship with arsenic exposure has not yet been clearly documented. The aim of our study was to investigate the dose-response relationship between arsenic exposure and major serum enzyme marker activity associated with LFTs in the population living in arsenic-endemic areas in Bangladesh.
Methods
A total of 200 residents living in arsenic-endemic areas in Bangladesh were selected as study subjects. Arsenic concentrations in the drinking water, hair and nails were measured by Inductively Coupled Plasma Mass Spectroscopy (ICP-MS). The study subjects were stratified into quartile groups as follows, based on concentrations of arsenic in the drinking water, as well as in subjects' hair and nails: lowest, low, medium and high. The serum hepatic enzyme activities of alkaline phosphatase (ALP), aspartate transaminase (AST) and alanine transaminase (ALT) were then assayed.
Results
Arsenic concentrations in the subjects' hair and nails were positively correlated with arsenic levels in the drinking water. As regards the exposure-response relationship with arsenic in the drinking water, the respective activities of ALP, AST and ALT were found to be significantly increased in the high-exposure groups compared to the lowest-exposure groups before and after adjustments were made for different covariates. With internal exposure markers (arsenic in hair and nails), the ALP, AST and ALT activity profiles assumed a similar shape of dose-response relationship, with very few differences seen in the higher groups compared to the lowest group, most likely due to the temporalities of exposure metrics.
Conclusions
The present study demonstrated that arsenic concentrations in the drinking water were strongly correlated with arsenic concentrations in the subjects' hair and nails. Further, this study revealed a novel exposure- and dose- response relationship between arsenic exposure metrics and serum hepatic enzyme activity. Elevated serum hepatic enzyme activities in the higher exposure gradients provided new insights into arsenic-induced liver toxicity that might be helpful for the early prognosis of arsenic-induced liver diseases.
doi:10.1186/1476-069X-10-64
PMCID: PMC3146918  PMID: 21740555
22.  Hypothetical mode of action of earthworm extract with hepatoprotective and antioxidant properties 
The hepatoprotective potential of earthworm extract (EE) (Lampito mauritii, Kinberg) was evaluated against paracetamol-induced liver injury in Wistar albino rat, in comparison with silymarin, the standard hepatoprotective drug. We observed a reduction in liver antioxidants, such as glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) and in serum total protein, and an increase in serum alkaline phosphatase (ALP), serum aspertate aminotranferase (AST), serum alanine aminotranferase (ALT), bilirubin and liver thiobarbituric acid reactive substances (TBARS) due to liver injury in the paracetamol-administered rats (2 g/kg). On the contrary, increased activities of liver GSH, SOD, GPx, CAT and serum total protein level, and decrease in the contents of serum ALP, AST, ALT, bilirubin and liver TBARS were observed in rats administered with different doses of EE (100, 200 and 300 mg/kg), which are similar to the activities of hepatoprotective drug silymarin (150 mg/kg). The mode of action of EE as evidenced by the above parameters may suggest that EE, on the one hand, prevents the formation of the reactive oxygen groups, or scavenges these groups, thereby preventing the damage on the hepatic cells, and, on the other hand, modulates the genes responsible for synthesis of antioxidant enzymes such as GPx, CAT and SOD in liver tissue and decreases the serum enzymatic activities such as ALP, AST and ALT.
doi:10.1631/jzus.B0720194
PMCID: PMC2225496  PMID: 18257136
Earthworm extract; Hepatoprotective potential; Antioxidant; Paracetamol; Lipid peroxidation
23.  Evaluation of Metabolic Enzymes in Response to Excel Mera 71, a Glyphosate-Based Herbicide, and Recovery Pattern in Freshwater Teleostean Fishes 
BioMed Research International  2014;2014:425159.
Metabolic enzymes such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were evaluated in Indian teleostean fishes, namely, Anabas testudineus (Bloch) and Heteropneustes fossilis (Bloch), for an exposure to 30 days of Excel Mera 71 (17.2 mg/L), a glyphosate formulation, and subsequent depuration under Liv.52, a plant extract at a dose of 187.5 mg/d/250 L for the same period in the same tissues under laboratory condition. ALT activity was significantly increased (P < 0.05) in all the tissues and raised up to 229.19% in liver of A. testudineus (229.19%) and 128.61% in liver of H. fossilis. AST also increased significantly (P < 0.05) and was maximum in liver of H. fossilis (526.19%) and minimum in gill of A. testudineus (124.38%). ALP activity was also raised highly in intestine of H. fossilis (490.61%) but was less in kidney of H. fossilis (149.48%). The results indicated that Excel Mera 71 caused alterations in the metabolic enzymatic activities in fish tissues and AST showed the highest alteration in both the fishes, while lowest in ALP and ALT in A. testudineus and H. fossilis, respectively. During depuration under Liv.52, all the enzyme activities came down towards the control condition which indicated the compensatory response by the fish against this herbicidal stress and it was in the following order: AST > ALT > ALP, in A. testudineus, while H. fossilis showed the following trend: ALT > AST > ALP. Therefore, these parameters could be used as indicators of herbicidal pollution in aquatic organisms and were recommended for environmental monitoring for investigating the mechanism involved in the recovery pattern.
doi:10.1155/2014/425159
PMCID: PMC4071806  PMID: 25013782
24.  Inhibitory potential of methanolic extracts of Aristolochia tagala and Curcuma caesia on hepatocellular carcinoma induced by diethylnitrosamine in BALB/c mice 
Context:
Aristolochia tagala (AT) and Curcuma caesia (CC) have been used traditionally by local herbal practitioners for cancer treatment and as chief ingredients of many polyherbal formulations for various types of ailments. However, there is void in scientific study to evaluate their anti-cancer property.
Aims:
The aim of this study was to evaluate the anti-carcinogenic properties of the crude methanolic extracts of roots of AT and rhizomes of CC in BALB/c mice exposed to a hepatocarcinogen, diethylnitrosamine (DEN).
Settings and Design:
(I) Toxicity of herbal plant extracts (HPE); (II) Anticancer studies; (III) Histological studies; and (IV) Biochemical studies.
Materials and Methods:
To evaluate the effects of these two HPE either alone or following DEN exposure, serum transaminases (aspartate aminotransferase [AST], alanine aminotransferase [ALT]), alkaline phosphatase (ALP), and cancer marker enzyme acetylcholine esterase (AChE) were assayed in mice. In addition, histological study was also carried out under similar conditions. The antioxidant potentials of the HPE were evaluated by monitoring the activity of antioxidant enzymes and metabolites, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione (GSH).
Statistical Analysis Used:
Statistical analysis was performed by GraphPad Prism 6 Software using one-way analysis of variance followed by the Tukey's multiple comparisons test. Significance was set at P < 0.05.
Results:
Our findings show that DEN administration elevated AST, ALT, ALP, and AChE activities. CC or AT extracts attenuated the increased activities of these marker enzymes. The activities of antioxidant enzymes, which were decreased following DEN administration, were significantly increased in mice treated with CC or AT.
Conclusions:
The present study clearly documents anticarcinogenic and antioxidant properties of AT and CC in DEN-induced mouse liver cancer in vivo.
doi:10.4103/1477-3163.133520
PMCID: PMC4141361  PMID: 25191135
Antioxidant; Aristolochia tagala; cancer; Curcuma caesia; diethylnitrosamine; liver
25.  Antiurolithiatic and antioxidant activity of Hordeum vulgare seeds on ethylene glycol-induced urolithiasis in rats 
Indian Journal of Pharmacology  2012;44(6):672-677.
Objective:
The objective was to investigate the antiurolithiatic and antioxidant activity of ethanolic extract of Hordeum vulgare seeds (EHV) on ethylene glycol-induced urolithiasis in Wistar albino rats.
Materials and Methods:
Urolithiasis was produced in Wistar albino rats by adding 0.75% v/v ethylene glycol (EG) to drinking water for 28 days. The ethanolic extract of Hordeum vulgare seeds (EHV) was assessed for its curative and preventive action in urolithiasis. In preventive treatment, the EHV given from 1st day to 28th day, while in the curative regimen, the EHV was given from 15th day to 28th day. Various renal functional and injury markers such as urine volume, calcium, phosphate, uric acid, magnesium, urea, and oxalate were evaluated using urine, serum, and kidney homogenate. Antioxidant parameters such as lipid peroxidation, superoxide dismutase, and catalase were also determined.
Results:
The EHV treatment (both preventive and curative) increased the urine output significantly compared to the control. The EHV treatment significantly reduced the urinary excretion of the calcium, phosphate, uric acid, magnesium, urea, and oxalate and increased the excretion of citrate compared to EG control. The increased deposition of stone forming constituents in the kidneys of calculogenic rats were significantly lowered by curative and preventive treatment with EHV. It was also observed that the treatment with EHV produced significant decrease in lipid peroxidation, and increased levels of superoxide dismutase and catalase.
Conclusion:
These results suggest the usefulness of ethanolic extract of Hordeum vulgare seeds as an antiurolithiatic and antioxidant agent.
doi:10.4103/0253-7613.103237
PMCID: PMC3523490  PMID: 23248392
Antioxidant; ethylene glycol; Hordeum vulgare; urolithiasis

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