姜黄素与阿米洛利联用对大鼠肝星状细胞氧应激所致纤维化的抑制作用

目的：探讨钠氢交换抑制剂阿米洛利(Amibride)与抗氧化剂姜黄素(curcumin，Cur)联用对大鼠肝星状细胞(hepatic stellate cells，HSC)氧应激所致纤维化的作用。方法：HSC与三醋酸亚硝酸铁(FeNTA)共同培养。以MTY法观察阿米洛利、姜黄素对HSC增殖的效应。LDH法观察阿米洛利、姜黄素对HSC的毒性作用。以：ELISA法测定细胞培养液中Ⅰ型胶原的水平。以试剂盒推荐方法分别测定细胞培养液中丙二醛(MDA)、谷胱苷肽(GSH)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)水平。结果：HSC与FeNTA共同培养，细胞内MDA、GSH水平明显升高，GSH-Px、SOD活力明显降低。FeNTA与HSC共同培养，能明显促进：HSC增殖和提高细胞培养上清中Ⅰ型胶原的水平。阿米洛利、姜黄素可以逆转上述所有FeNTA所致效应，且联用组优于单用组。结论：作用于HSC不同靶点的抗纤维化药物姜黄素、阿米洛利合用优于单用。

Inhibitive Effect of Curcumin and Amiloride on the Fibrosis of Rat Hepatic Stellate Cells Induced by Oxidative Stress

OBJECTIVE: To investigate the effects of antioxidant curcumin and Na+/H+ exchanger inhibitor amiloride on the fibrosis of rat hepatic stellate cells (HSC) induced by oxidative stress in vitro. METHODS: HSC were incubated with 0.1 mmol/L ferric nitrilotriacetate complex (FeNTA). MTT colorimetry was used for assaying proliferation of HSC. Cytotoxicity was measured by LDH colorimetry. Collagen type I accummulation in the culture media was measured by ELISA. Intracellular malonildialdehyde (MDA), SOD, GSH and GSH-Px levels in the culture media were measured by respective reagent boxes. RESULTS: HSC incubation with FeNTA resulted in a significant production of intracellular MDA and GSH, associated with decrease of SOD and GSH-PX activity. Exposure of HSC to FeNTA significantly enhanced the number of proliferating HSC and collagen type I levels in the culture medium. All these effects were reversed by the antioxidant curcumin and by the Na+/H+ exchanger inhibitor amiloride. Combining curcumin with amiloride had better effects on the fibrosis of rat hepatic stellate cells induced by oxidative stress than curcumin or amiloride. CONCLUSION: The effects of combination of antifibrosis drugs curcumin and amiloride on different targets of HSC are superior to curcumin or amiloride.