【摘要】 目的 :探讨氧化苦参碱 (OM )阻断苯巴比妥钠 (PB)诱导的肝细胞凋亡的机制。方法 :10 5只雄性昆明种小鼠随机分为 5组。空白对照组 :腹腔注射生理盐水 ,连续 7d ,停药 12h后处死。PB组 :5 g/LPB 75mg/kg ,1次/d ,腹腔注射 ,连续 7d ,停药后 12h处死。PB撤除组 :5 g/LPB 75mg/kg ,1次 /d ,腹腔注射 ,连续 6d ,停药后 36h处死。OM治疗Ⅰ组 :首先腹腔注射 5g/LPB 75mg/kg ,1次 /d ;第 6d后腹腔注射OM 15 0mg/kg ,3次 /d ,至停PB后 36h ,将动物处死。OM治疗Ⅱ组 :首先腹腔注射 5g/LPB 75mg/kg ,1次 /d ,第 6d后腹腔注射OM 75mg/kg ,3次 /d ,至停PB后 36h ,将所有动物脱颈处死 ,取出肝脏 ,体积分数为 4 %多聚甲醛固定 ,石蜡包埋。用Tunel法检测肝细胞凋亡 ,用免疫组化和原位杂交技术检测bcl 2、bax基因的表达。结果 :仅PB撤除组Tunel法检测到细胞凋亡。Bcl 2蛋白在OM治疗Ⅰ、Ⅱ组的表达较PB撤除组显著增强 (P <0 .0 5 ) ,在OM治疗Ⅰ组和Ⅱ组之间差异无统计学意义。Bax蛋白在PB撤除组表达较其他各组显著增强 (P <0 .0 5 ) ,而在其他各组之间差异无统计学意义(P >0 .0 5 )。bcl 2、bax基因表达的mRNA阳性信号强度与Bcl 2、Bax蛋白免疫组化反应的强度一致。结论 :氧化苦参碱可能通过上调bcl 2基因的表达来阻断肝?更多还原

【Abstract】 Aim:To explore the mechnism of OM in inhibition of hepatocyte from apoptosis induced by PB.Methods: 105 Kumming mice were divided randomly into 5 groups. The TUNEL assay was used to detect the apoptosis of hepatocytes, the immunohistochemistry and in situ hybridization techniques were used to detect the gene expression of bcl 2 and bax in hepatocytes, respectively.Results:The apoptosis detected by Tunel method is positive in the PB withdrawal group, the other groups is negative.The immunohistochemistry staining showed that the positive immunoreactivity (IR) of Bcl 2 and Bax appeared as brownish granules and was localized at cytoplasm and nuclear envelope. The Bcl 2 IR was negative in the blank control group and the PB group, the level of Bcl 2 IR in the OM Ⅰ and OM Ⅱ treatment groups was significantly higher than that of the PB withdrawal group ( P <0.05), but there were no significant difference between the OM Ⅰ and OM Ⅱ treatment groups ( P >0.05). The level of Bax IR in the PB withdrawal group was higher significantly than that of the other groups ( P <0.05), there were no significant difference between the other groups ( P >0.05).In situ hybridization result. The positive intensity of mRNA signal was coincident to that of the protein immunoreactivity in all groups, the positive signal of bcl 2 mRNA is strong in the OM Ⅰ and the OM Ⅱ treatment groups, the positive signal of bax mRNA is strong in the PB withdrawal groups. Conclusions:OM could inhibit hepatocyte from apoptosis induced by PB by up regulate the expression of bcl 2 gene.更多还原