达肝清对小鼠肝细胞异常凋亡影响.docVIP

达肝清对小鼠肝细胞异常凋亡影响 　【摘要】 目的探讨中药复方达肝清对苯巴比妥钠诱导的小鼠肝细胞异常凋亡的影响，进一步揭示其抗肝损伤的作用机理。方法采用撤除苯巴比妥钠（PB）诱导 小鼠肝细胞凋亡的模型；通过流式细胞检测技术，TUNEL原位检测法，HE常规染色等方法，以肝细胞凋亡率、肝组织形态、原位细胞TUNEL反应、肝/体重比为指标，观察不同剂量达肝清对PB诱导肝细胞凋亡的影响。结果流式细胞仪分析表明，达肝清高、中剂量组肝细胞凋亡率明显低于凋亡对照组；达肝清高剂量组肝/体重比回落明显低于凋亡对照组；组织学染色及TUNEL反应，凋亡对照组可见凋亡小体，凋亡现象明显，凋亡细胞明显多于达肝清高、中剂量组。结论达肝清能明显抑制苯巴比妥钠诱导的肝细胞凋亡，进一步提示抑制肝细胞异常凋亡可能为其抗肝损伤的机理之一。 【关键词】 达肝清；肝细胞凋亡；流式细胞术；原位末端缺口标记 　　 　　Abstract：ObjectiveTo study the effects of Chinese herb compound Daganqing on apoptosis caused by phenobarbital withdrawal of liver cells in mice, and to research the mechanism of its preventing acute liver injury. MethodsThe apoptosis model of liver cells in mice caused by phenobarbital withdrawal was used, FCM, TUNEL, and HE routine dye, were performed to check the changes in apoptotic cells. The rate of hepatocyte apoptosis, the hepatic histopathologic changes, tunnel change and liver body weight ratio were observed to study the effects of Daganqing in different dosages on apoptosis caused by phenobarbital withdrawal of liver cells. ResultsFlow cytometry analysis revealed that the apoptotic rate of liver cells and liver body weight ratio of Daganqing high and middle dose groups were obviously lower than positive group. By the analysis of TUNEL, and HE routine dye, apoptotic bodies and cell apoptosis were more conspicuous, and the number of apoptotic cell was more in positive group than other groups. ConclusionChinese herb compound Daganqing in the dosages of 34 g/kg，17 g/kg can obviously inhibit the liver cell apoptosis caused by phenobarbital withdrawal, and further reveal that inhibiting the liver cell apoptosis maybe one of the mechanisms of Daganqing preventing acute liver injury. 　　Key words：Chinese herb compound Daganqing；Apoptosis of liver cells； FCM； TUNEL 　　达肝清是由三姐妹、黄根等药物组成的中药复方，具有清热解毒、益气健脾、利湿化瘀等功效，临床上主要用于治疗急、慢性病毒性肝炎，疗效显著。研究已表明达肝清具有保肝降酶,抗乙肝病毒等作用［1］,对急、慢性肝损伤具有明显的保护作用［2］,同时对2215细胞分泌的乙型肝炎病毒表面抗原（HBsAg）和e抗原（HBeAg）有抑制作用［3］。本实验采用撤除苯巴比妥钠诱导肝细胞凋亡模型，观察达肝清对肝细胞异常凋亡的影响，以进一步揭示其治疗病毒性