抗氧化剂对大鼠肝星形细胞增殖和凋亡影响的实验研究

Effect of antioxidant on hepatic stellate cell proliferation and apoptosis during recovery from liver fibrosis

目的 了解膳食维生素E和硒对大鼠肝纤维化恢复期肝星形细胞 (HSC)增殖和凋亡的影响。方法 采用腹腔内注射 5 0 %CCl4 制备大鼠肝纤维化模型 ,在饲料中添加适量维生素E(2 5 0mg kg饲料 )和Se(0 . 2mg kg饲料 )进行营养干预 ,在最后一次注射后 3、7、14、2 8天 (恢复期 )各处死 3只大鼠取其肝组织 ,用HE和Siriusred染色结合图象分析检测肝纤维化指标 ,用α SMA免疫组化方法检测激活的HSC细胞 ,用原位凋亡 (TUNEL)技术和α SMA免疫组化双标染色检测HSC凋亡。结果 在恢复期各时间点 ,病理对照组和抗氧化干预组激活的HSC数量呈逐步下降趋势 ,同时 ,从恢复期第 7d开始 ,HSC凋亡细胞数以及肝组织内的胶原量亦同步下降 ;在同一时间点 ,干预组激活的HSC数量低于病理对照组 ,而HSC凋亡数和凋亡率均高于病理对照组。结论 饲料中添加适量维生素E和硒能减轻肝纤维化的程度 ,并可能使恢复期HSC凋亡增加 ,激活HSC数量减少。

Objective Effects of dietary supplementation of vitamin E and selenium on prolif eration and apoptosis of activatd hepatic stellate cell(HSC) were investigated in the rat model of liver fibrosis induced by intraperitoneal injection with CCl 4.Methods Activatd HSC was determined by α-smooth muscle ac tin immunohistochemistry st aining and apoptotic HSC determined by dual staining both of the terminal deoxy nucleotidyl transferase UTP nick end labeling(TUNEL) and of α-smooth muscle ac t in immunohistochemistry. Result During fibrosis reco very,the numb er of activated HSCs both in pathological group and in intervention group went d own gradually,meanwhile, both the number of apoptotic HSCs and the collegen live r also descend little by little.These data confirmed that HSCs had the core effe ct on liver fibrogenesis and apoptosis may be a major factor regulating HSCs num bers during the injury-fibrosis-recovery sequence.At each time point, the numb er of actived HSCs in pathological group is more than intervention group, while a poptic HSCs are less,which suggested dietary supplement with antioxidative nutri ents had effect on HSC apoptosis but more studies are necessary to make the mech anism clearer.Conclusion dietary supplement with prope r vitamin E and selenium can effectively lighten the hepatic fibrosis and promot e the recovery of hepatocyte and the degradation of the existing collagens,ie,it is beneficial to the recovery of hepatic fibrosis.