Protective effects of silybin on human umbilical vein endothelial cell injury induced by H2O2 in vitro.
Oxidative injury induces cellular and nuclear damages that lead to cell injury. Agents or antioxidants that can inhibit production of reactive oxygen species can prevent injury. We tested the hypothesis that silybin can inhibit H2O2-induced injury in human umbilical vein endothelial cells. Eighteen hours of treatment with 750 micromol l(-1) H2O2 significantly stimulated expression of caspase-3 and cell apoptosis. In addition, it is observed that H2O2 increased the amounts of malondialdenhyde (MDA), the dehydrogenase (LDH) leakage, and decreased the activity of GSH-Px and NO contents in ECV-304 cells. In the H2O2 apoptosis model, the addition of 6.25-25 mg/L of silybin, which has in vitro radical scavenging activity, partially restored cell viability with a reduction in H2O2-induced apoptotic DNA damage, and decreased the expression of caspase-3. Moreover, it decreased other H2O2-induced damage in a concentration-dependent manner. The endothelial cell apoptosis was detected by AO/EB dual staining as well as flow cytometry, and the activity of pro-apoptotic factor caspase-3 was detected by immunocytochemical method. Our results suggest that the antioxidant, silybin, protects ECV-304 cells against H2O2-induced injury probably through its antioxidant activity, increasing the NO content, the activity GSH-Px and inhibiting signaling pathways mediated by caspase-3.
Wang
Yun-Kai
YK
Department of Cardiology, The Affiliated Hospital of Zhejiang Traditional Medicine College, Hangzhou 310006, PR China.
Hong
Ya-jun
YJ
Huang
Zhao-Quan
ZQ
D000975:Antioxidants / Q000494:pharmacology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects*; D004847:Epithelial Cells / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects; D005920:Glomerular Mesangium / Q000166:cytology / Q000187:drug effects; D005979:Glutathione Peroxidase / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000097:blood; D008315:Malondialdehyde / Q000097:blood; D009569:Nitric Oxide / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology*; D011794:Quercetin / Q000494:pharmacology; D000077385:Silybin; D012838:Silymarin / Q000494:pharmacology; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects
D000975:Antioxidants; D016877:Oxidants; D012838:Silymarin; D009569:Nitric Oxide; D000077385:Silybin; D008315:Malondialdehyde; D011794:Quercetin; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D005979:Glutathione Peroxidase; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases