蒿鳖养阴软坚方对肝纤维化大鼠抗氧化损伤与诱导凋亡的作用  

Effect of HBYYRJ Herbal Extract on Anti-Oxidation Injury and Induction of Apoptosis in Rat Models of Hepatic Fibrosis

作　　者：刘红艳[1,2] 方步武[1] 率红莉[1,3] 邢伟[1,4] 刘洁莹[1] 赵敏[1] 郭娟[1] 

机构地区：[1]天津医科大学药理教研室,300070 [2]天津市南开医院 [3]天津市安宁医院 [4]天津医科大学总医院

出　　处：《天津医药》2010年第10期884-887,共4页Tianjin Medical Journal

基　　金：天津市应用基础研究计划重点项目(项目编号:06YFJZJC02900)

摘　　要：目的:探讨蒿鳖养阴软坚方抗氧化损伤和诱导凋亡而预防CCl4复合因素所致肝纤维化的作用。方法:大鼠80只按体质量分层随机抽样法分为7组。正常组6只,模型组14只,蒿鳖养阴软坚方低剂量组12只,中剂量组12只,高剂量组12只,复方鳖甲软肝片组12只和秋水仙碱组12只。采用复合因素CCl4注射、高脂高胆固醇玉米饼饲料、间断少量乙醇灌胃制备大鼠肝纤维化模型,以蒿鳖养阴软坚方粗提物预防用药,HE染色及胶原染色观察肝组织形态,基于T4 DNA连接酶的ApopTag ISOL法检测凋亡,免疫组化SABC法检测α-平滑肌肌动蛋白(α-SMA),水解氧化比色法测肝组织羟脯氨酸(Hyp),比色法测血清丙二醛(MDA)。结果:与模型组相比,蒿鳖养阴软坚方中剂量与高剂量组具有显著降低肝纤维化程度分期和胶原蛋白含量的作用(P<0.01)。高剂量组可降低丙二醛(MDA)水平(P<0.05),减低α-SMA表达。中剂量组可诱导间质细胞凋亡(P<0.01),但抑制肝细胞凋亡(P<0.01)。结论:蒿鳖养阴软坚方通过抗氧化损伤、抑制HSC活化、促进活化的HSC凋亡等而发挥抗肝纤维化的作用。Objective：To evaluate the effect of HBYYRJ herbal extract on anti-oxidation injury and induction of apoptosis in rat models of hepatic fibrosis caused by CCl4 composed factor.Methods：Eighty rats were divided into 7 groups by randomized block of weight,normal group（n = 6）,model group（n = 14）,low dose HBYYRJ group（n = 12）,middle dose group（n = 12）,high dose group（n = 12）,composed formula treatment group（n = 12） and colchicine treatment group（n = 12）.The rat models of hepatic fibrosis were established by injection of CCl4 composed factor,and by feeding a high lipid and high cholesterol corn diet and intermittent minimal alcohol gastric perfusion.HBYYRJ herbal crude extract was used to prevent hepatic fibrosis.Histological observation of the liver tissue was done with HE stain and collagen stain.Apoptosis was tested with ApopTag ISOL on the basis of T4 DNA ligase.The value of α-smooth muscle actin（α-SMA） was determined with immunohistochemical SABC method.Hydroxyproline in liver tissue was tested by hydrolyzed oxidation colorimetry.The serum malondialdehyde（MDA） was tested with colorimetry.Results：As compared with the model group,the degrees of liver fibrosis and the contents of collagen were significantly reduced in the middle and high dose treatment groups（P 0.01）.The level of MDA and the expression of α-SMA were significantly decreased in high dose treatment group than those of model group（P 0.05）.The apoptosis of interstitial cells was induced（P 0.01）,but apoptosis of hepatic cells was inhibited,in middle dose group（P 0.01）.Conclusion：HBYYRJ herbal extract can prevent and inhibit liver fibrosis through the effects of anti-oxidation injury,inhibiting activation of hepatic stellate cells and promoting apoptosis of active hepatic stellate cells.

关 键 词：四氯化碳 肝硬化 丙二醛 肌动蛋白类 脂质过氧化作用 细胞凋亡 大鼠 Wistar 

分 类 号：R259.752[医药卫生—中西医结合]