目的:观察复方鳖甲软肝方药物血清对离体SD大鼠肝星形细胞(hepaticstellate cells,HSC)结蛋白、突触素表达的影响,以探讨其抗肝纤维化作用机制.方法:将成年健康雄性SD大鼠75只,随机分为正常对照组、模型组、高、中、低剂量药物组共5组,并制备各组血清.采用大鼠HSC分离技术,将分离得到的HSC分别添加各组血清培养24,48,72 h后,免疫组化检测各组HSC的结蛋白、突触素表达,并经图像分析系统作定量分析.结果:培养24 h正常对照组、高、中、低剂量药物组大鼠HSC结蛋白、突触素的表达(A值)(78.93±18.56,396.29±78.56;58.95±5.21,138.92±20.21;63.83±12.97,190.87±52.97;75.46±26.72,284.54±66.72)与模型组(119.69±23.84,528.79±26.84)比较,差异有显著性意义(P＜0.05);培养48,72 h各组结蛋白、突触素的表达(A值)与模型组比较,差异也有显著性意义(P＜0.05).除低剂量药物组外的其余各组大鼠HSC结蛋白、突触素表达与正常对照组比较,差异有显著性意义(P＜0.05).结论:复方鳖甲软肝方能显著抑制HSC结蛋白、突触素的表达,其抗肝纤维化作用机制可能与药物抑制HSC活化有关.
AIM:To observe the influence of drug serumof fufang biejia ruangan fang on the expressions of desmin and synapsin of hepatic stellate cell(HSC) in vitro,so as to discuss its mechanism of anti hepatic fibrosis of SD rats. METHODS:A total of 75 adult healthy male SD rats were randomly divided into normal control group,model group,high, middle and low dose groups(fufang biejia ruangan fang),and the serum was prepared in all the groups.The HSCs were separated with the HSC separating technique,and then cultured with different serum for 24,48 and 72 hours,then the expressions of desmin and synapsin in HSC were detected by using immunohistochemistry,and then the quantitative analysis was performed by using image analysis system. RESULTS:After culture for 24 hours,the expressions of desmin and synapsin of HSC(absorbance,A) in the normal control group (78.93±18.56,396.29±78.56),high dose drug group(58.95±5.21,138.92±20.21),middle dose group (63.83±12.97,190.87±52.97)and low dose drug group(75.46±26.72,284.54±66.72) were significantly different from those in the model group(119.69±23.84,528.79±26.84)(P< 0.05),and the A values were all significantly different from those in the model group 48 and 72 hours after culture(P< 0.05);the A values in all groups except the low dose drug group had significant differences with those in the normal control group(P< 0.05).CONCLUSION:Fufang biejia ruangan fang can significantly inhibit the expressions of desmin and synapsin of HSC.The mechanism for anti hepatic fibrosis by fufang biejia ruangan fang may be related with the anti activation of HSC by drug.
Chinese Journal of Clinical Rehabilitation