Negundoside, an irridiod glycoside from leaves of Vitex negundo, protects human liver cells against calcium-mediated toxicity induced by carbon tetrachloride.
OBJECTIVE
To evaluate the protective effect of 2'-p-hydroxybenzoylmussaenosidic acid [negundoside (NG), against carbon tetrachloride (CCl(4))-induced toxicity in HuH-7 cells.


METHODS
CCl(4) is a well characterized hepatotoxin, and inducer of cytochrome P450 2E1 (CYP2E1)-mediated oxidative stress. In addition, lipid peroxidation and accumulation of intracellular calcium are important steps in the pathway involved in CCl(4) toxicity. Liver cells (HuH-7) were treated with CCl(4), and the mechanism of the cytoprotective effect of NG was assessed. Silymarin, a known hepatoprotective drug, was used as control.


RESULTS
NG protected HuH-7 cells against CCl(4) toxicity and loss of viability without modulating CYP2E1 activity. Prevention of CCl(4) toxicity was associated with a reduction in oxidative damage as reflected by decreased generation of reactive oxygen species (ROS), a decrease in lipid peroxidation and accumulation of intracellular Ca(2+) levels and maintenance of intracellular glutathione homeostasis. Decreased mitochondrial membrane potential (MMP), induction of caspases mediated DNA fragmentation and cell cycle arrest, as a result of CCl(4) treatment, were also blocked by NG. The protection afforded by NG seemed to be mediated by activation of cyclic adenosine monophosphate (cAMP) synthesis and inhibition of phospholipases (cPLA2).


CONCLUSIONS
NG exerts a protective effect on CYP2E1-dependent CCl(4) toxicity via inhibition of lipid peroxidation, followed by an improved intracellular calcium homeostasis and inhibition of Ca(2+)-dependent proteases.
Tasduq
Sheikh A
SA
Experimental Toxicology Lab, Division of Pharmacology, Indian Institute of Integrative Medicine, CSIR, Canal Road, Jammu 180001, Jammu and Kashmir, India. tasduq11@gmail.com
Kaiser
Peerzada J
PJ
Gupta
Bishan D
BD
Gupta
Vijay K
VK
Johri
Rakesh K
RK
D000975:Antioxidants / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D002251:Carbon Tetrachloride / Q000633:toxicity*; D053148:Caspase 3 / Q000378:metabolism; D061945:Caspase Inhibitors; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D000242:Cyclic AMP / Q000378:metabolism; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D019610:Cytoprotection; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D006801:Humans; D057889:Iridoid Glycosides; D039823:Iridoids / Q000302:isolation & purification / Q000494:pharmacology*; D015227:Lipid Peroxidation / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D054510:Phospholipases A2, Cytosolic / Q000037:antagonists & inhibitors / Q000378:metabolism; D018515:Plant Leaves; D017382:Reactive Oxygen Species / Q000378:metabolism; D012838:Silymarin / Q000494:pharmacology; D020950:Vitex / Q000737:chemistry
D000975:Antioxidants; D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D057889:Iridoid Glycosides; D039823:Iridoids; D017382:Reactive Oxygen Species; D012838:Silymarin; C532410:negundoside; D045304:Cytochromes c; D002251:Carbon Tetrachloride; D000242:Cyclic AMP; D019392:Cytochrome P-450 CYP2E1; D054510:Phospholipases A2, Cytosolic; C505407:CASP3 protein, human; D053148:Caspase 3; D005978:Glutathione; D002118:Calcium