论文部分内容阅读
目的研究小鼠肾缺血再灌注后不同时间点热休克蛋白HSP90α和miRNA-144的表达变化。方法用动脉夹夹闭小鼠双侧肾蒂45min建立肾缺血再灌注损伤模型,测定再灌注4、8、12、24、48h组及假手术组血清中肌酐(Scr)、尿素氮(BUN)水平,光镜下观察肾组织形态学改变,采用定量RT-PCR的方法检测肾缺血再灌注各时间点HSP90αmRNA和miRNA-144的表达变化,Western blot法检测HSP90α蛋白的表达变化。结果 HE染色显示假手术组小鼠的肾组织结构及肾功能正常,再灌注组可见肾小管管腔扩张,肾小管上皮细胞有不同程度的肿胀、坏死、刷状缘消失,管腔内有管型,肾间质充血水肿,组织损伤以24h最重;肾功能指标显示再灌注组与假手术组相比,Scr、BUN水平随再灌注时间延长而升高,24h达峰值(P<0.01)。定量RTPCR和Western blot法检测结果显示,再灌注4h后HSP90α的mRNA和蛋白表达开始逐渐增强,mRNA在12h达高峰,蛋白水平在24h达高峰(P<0.01);miRNA-144在再灌注后表达降低(P<0.01)。结论 HSP90α在缺血再灌注损伤的肾组织中表达增高,提示其可能参与肾缺血再灌注损伤的保护作用;miRNA-144可能为调控HSP90α功能的潜在miRNA。
Objective To study the expression of heat shock protein HSP90α and miRNA-144 at different time points after renal ischemia-reperfusion in mice. Methods The model of renal ischemia-reperfusion injury was established by clamping the bilateral renal pedicles 45min with the artery clamp. Serum creatinine (Scr), blood urea nitrogen (BUN ). The morphological changes of renal tissue were observed under light microscope. The expression of HSP90αmRNA and miRNA-144 at different time points after renal ischemia-reperfusion were detected by quantitative RT-PCR. The expression of HSP90α protein was detected by Western blot. Results HE staining showed that the kidneys of mice in sham-operation group had normal renal structure and renal function. Renal tubular dilatation was observed in reperfusion group. Tubular epithelial cells were swollen and necrotic in some degree and disappeared in brush border, Type and renal interstitial hyperemia and edema, the histological injury was the heaviest in 24 h. The renal function indexes of Scr and BUN increased with the reperfusion time prolonged and peaked at 24 h (P <0.01) . The results of quantitative RT-PCR and Western blot showed that the mRNA and protein expression of HSP90α began to increase after 4h of reperfusion, reached its peak at 12h and peaked at 24h (P <0.01). The expression of miRNA-144 was detected after reperfusion Decreased (P <0.01). Conclusions HSP90α expression is increased in renal tissue with ischemia-reperfusion injury, suggesting that HSP90α may be involved in the protective effect of renal ischemia-reperfusion injury. MiRNA-144 may be a potential miRNA for regulating the function of HSP90α.