【摘要】 目的探讨梗阻性黄疸引起肾损伤过程中细胞凋亡的意义。方法32只雄性SD大鼠随机均分为2组,假手术组(SO组)只暴露腹腔但不结扎胆管,胆总管结扎组(BDL组)以双重结扎胆管中间离断方法制成梗阻性黄疸大鼠模型。于术后行光镜下肾脏病理学检查,琼脂糖凝胶电泳检测肾细胞DNA的断裂形式,测定血清中D-Bil、TBA、GOT、GPT、Cr和BUN的含量,检测尿β2-微球蛋白,流式细胞术检测肾脏细胞凋亡率,免疫组化法检测肾组织凋亡抑制基因bcl-2蛋白的表达。结果BDL组大鼠肾脏细胞凋亡率明显高于SO组(P<0.05),出现细胞凋亡特征性DNA梯状带;其肾脏细胞bcl-2蛋白的表达阳性细胞率明显高于SO组(P<0.01),并于术后7d达高峰,14d时bcl-2表达阳性细胞率开始下降。结论梗阻性黄疸大鼠肾功能损害过程中存在肾脏细胞凋亡,梗阻性黄疸可反馈性启动肾小管上皮细胞凋亡抑制基因bcl-2的表达。更多还原

【Abstract】 Objective To investigate the effect of renal cell apoptosis induced by obstructive jaundice on the expression of bcl-2 in rats, and to explore the mechanism of renal impairment induced by obstructive jaundice. Methods Thirty-two male SD rats were randomly divided into 2 groups: SO group and BDL group. The rats in SO group received sham operation. Bile ducts of rats in BDL group were ligated. Pathology of kidneys was observed under the microscope. The levels of D-Bil, TBA, GOT, GPT, Cr and BUN in serum and β2-MG in urine were measured. The apoptotic rate of renal cells was calculated by flow cytometry and the forms of DNA fragmentation in renal cells were detected by agarose gel electrophoresis. The expression of inhibitory gene bcl-2 in the renal tissues was detected by immunohistochemistry. Results The color of urine in BDL group became dark yellow in day 2 after operation; The ears, tails and the muscle of abdominal wall and splanchnic organs, such as liver and kidney, also became yellow and swollen in day 7. The D-Bil, TBA, GOT, GPT, BUN of serum and β2 -MG of urine in BDL group were higher than those in SO group (P<0.05, P<0.01), and each value (except β2 -MG) in BDL group of 14 d was higher than that in BDL group of 7 d (P<0.05, P<0.01), respectively. The result of flow cytometry showed that the apoptotic rate of SO group and BDL (7 d and 14 d) group were (2.10±0.75)%, (18.17±0.86)% and (36.39±2.23)% respectively, there were significantly difference among them (P<0.05). The expression rate of bcl-2 of renal cell in BDL group of 7 d was higher than that in BDL group of 14 d. Conclusion Obstructive jaundice could induce apoptosis of the renal cells, and activate the expression of bcl-2 of the renal tubular epithelial cells in feedback, which may regulate the process of apoptosis.更多还原