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氧化苦参碱对苯巴比妥钠诱导的肝细胞凋亡小鼠肝细胞bcl-2、bax基因表达的影响

Effects of oxymatrine on expression of both bcl-2 and bax genes related to apoptotic in mouse hepatocytes

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【作者】 唐翠兰江河清孙长宇安敬军周舟

【Author】 TANG Cuilan, JIANG Heqing, ZHOU Zhou, SUN Changyu, HAN Mingguang 1)Department of infectious diseases, the First Affiliated Hospital, Zhengzhou university Zhengzhou 450052 2)Department of infectious diseases, the Third Affiliated Hospital of Xinxiang Medical College, Xinxiang 453003

【机构】 新乡医学院第三附属院传染科郑州大学第一附属医院传染科新乡医学院第三附属院传染科 新乡453003郑州450052新乡453003

【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?

【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.

【关键词】 氧化苦参碱肝细胞细胞凋亡bcl2苯巴比妥钠
【Key words】 oxymatrinehepatocyteapoptosisbcl 2phenobarbital
  • 【文献出处】 郑州大学学报(医学版) ,Journa of Henan Medical University , 编辑部邮箱 ,2002年06期
  • 【分类号】R285.5
  • 【被引频次】5
  • 【下载频次】143
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