Caspase-1介导的PARP1激活在肾小管间质纤维化中的作用及机制

发布时间：2018-05-05 23:58

本文选题：肾间质纤维化 + 单侧输尿管梗阻　；参考：《华中科技大学》2014年博士论文

【摘要】：目的：探讨caspase-1和PARP1在肾间质纤维化中的作用及其机制。方法：将C57BL/6J野生型小鼠和caspase-1-/-型小鼠分别制备单侧输尿管梗阻(uuo)模型,共分为4组：野生型假手术组(WT sham-operated);野生型模型组(WT UUO); caspase-1-/-型假手术组(KO sham-operated)和caspase-1-/-型模型组(KO UUO).7天后处死小鼠取肾组织标本,应用Western Blot检测caspase-7表达及激活水平、PARP1剪切与激活水平和α-SMA表达；免疫组化检测α-SMA和fibronectin的表达和分布；Masson染色检测胶原纤维的表达与分布；荧光实时定量PCR检测α-SMA mRNA表达水平。结果：(1)与WT sham-operated组相比,WT UUO组激活型caspase-7表达明显上调,差异具有统计学意义(P0.05)。KO UUO组与WT UUO组相比,caspase-7激活明显减少(P0.05),且α-SMA蛋白表达水平和mRNA表达下调(P0.05),而PARP1的剪切和激活并无明显变化。(2)免疫组化结果显示：与WT sham-operated组比,WT-UUO组α-SMA和fibronectin表达增加,而KOUUO组与WTUUO组相比,α-SMA和fibronectin表达明显减少(P0.05)。 Masson染色结果表明,KOUUO组与WTUUO组相比,胶原含量减少(P0.05)。结论：Caspase-1基因敲除可降低caspase-7的活性,并使α-SMA和fibronectin表达下调,但对PARP1的激活无明显影响。这提示caspase-1-caspase-7信号通路在肾纤维化中起重要作用。
[Abstract]:Objective: to investigate the role and mechanism of caspase-1 and PARP1 in renal interstitial fibrosis. Methods: C57BL/6J wild-type mice and caspase-1-r-type mice were used to establish unilateral ureteral obstruction models. The mice were divided into four groups: wild type sham-operated group (WT sham-operated) group; wild type model group (WWT UUO); caspase-1-r-type sham-operated group (KO sham-operated group) and caspase-1-r-type model group (Caspase-1-r-type model group). The expression and distribution of 伪 -SMA and 伪 -SMA were detected by Western Blot, the expression and distribution of 伪 -SMA and 伪 -SMA by immunohistochemistry, and the expression of 伪 -SMA mRNA by real-time fluorescence quantitative PCR. Results compared with WT sham-operated group, the expression of activated caspase-7 increased significantly in WT UUO group. Compared with WT UUO group, the activation of caspase-7 in KO UUO group was significantly lower than that in WT UUO group, and the expression level of 伪 -SMA protein and mRNA expression were down-regulated in P0.05, while the shear and activation of PARP1 had no significant change. The immunohistochemical results showed that: compared with WT sham-operated group, the expression of 伪 -SMA protein was lower than that of WT sham-operated group. The expression of 伪 -SMA and fibronectin increased in WT-UO group. The expression of 伪 -SMA and fibronectin in KOUUO group was significantly lower than that in WTUUO group. The results of Masson staining showed that the content of collagen in KUO group was lower than that in WTUUO group. Conclusion the knockout of the Caspase-1 gene can decrease the activity of caspase-7 and down-regulate the expression of 伪 -SMA and fibronectin, but has no effect on the activation of PARP1. This suggests that caspase-1-caspase-7 signaling pathway plays an important role in renal fibrosis.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R692.6

【共引文献】