基于凝血酶诱导的PAR-1/p38 MAPK/NF-κB通路探讨水蛭素干预特发性膜性肾病及其肾间质纤维化的机制研究

Idiopathic membranous nephropathy is a common cause of nephrotic syndrome, renal interstitial fibrosis is an important basis for the formation of end-stage renal disease. In this study, the incidence of idiopathic membranous nephropathy and its occurrence of interstitial fibrosis as the starting point, around the "thrombin can activate PAR-1 / p38 MAPK / NF-κB pathway mediated idiopathic membranous nephropathy Attack complex formation and renal interstitial fibrosis in early inflammatory cell infiltration and release of fibrogenic factors "in this hypothesis, combined with the characteristics of hirudin-specific binding to thrombin, through in vivo studies, the use of pathology, molecular biology Technology, on the pathogenesis of idiopathic membranous nephropathy after hirudin intervention and PAR-1, p38 MAPK, NF-κB and other related to the early cell pathways of renal interstitial fibrosis, promote fibrosis factor TGF-β1 and inflammatory factor ICAM- 1, ICAM-3, MCP-1, MCP-2, MCP-3 and cell surface markers a-SMA, E-Cadherin were observed to explore the pathogenesis of idiopathic membranous nephropathy and its renal Interventional effects of early cell signal transduction in fibrosis can provide new intervention targets and experimental basis for the research and development of idiopathic membranous nephropathy and anti-renal interstitial fibrosis.

特发性膜性肾病是肾病综合征的常见病因，肾间质纤维化是终末期肾病形成的重要基础。本研究以特发性膜性肾病发病及其肾间质纤维化发生为切入点，围绕“凝血酶可通过激活PAR-1/p38 MAPK/NF-κB细胞通路介导特发性膜性肾病膜攻击复合物形成及肾间质纤维化早期炎症细胞浸润和致纤维化因子的释放”这一假说，结合水蛭素特异性结合凝血酶的特点，通过在体实验研究，利用病理学、分子生物学技术，对水蛭素干预后特发性膜性肾病发病过程及肾间质纤维化早期细胞通路相关的PAR-1,p38 MAPK,NF-κB等，促纤维化因子TGF-β1和炎症因子ICAM-1,ICAM-3,MCP-1,MCP-2,MCP-3及细胞表面标志物a-SMA,E-Cadherin的表达进行观察，探讨水蛭素对于特发性膜性肾病发病过程及其肾间质纤维化早期细胞信号转导的干预效应，为研发治疗特发性膜性肾病、抗肾间质纤维化提供新的干预靶点和实验根据。

特发性膜性肾病是肾病综合征的常见病因，肾间质纤维化是终末期肾病形成的重要基础。本研究以特发性膜性肾病发病及其肾间质纤维化发生为切入点，围绕“凝血酶可通过激活PAR-1/p38 MAPK/NF-κB细胞通路介导特发性膜性肾病膜攻击复合物形成及肾间质纤维化早期炎症细胞浸润和致纤维化因子的释放”这一假说，结合水蛭素特异性结合凝血酶的特点，通过在体实验研究，利用病理学、分子生物学技术，探讨水蛭素对于特发性膜性肾病发病过程及其肾间质纤维化早期细胞信号转导的干预效应。结果表明：水蛭素可在一定程度上降低PHN大鼠24h尿蛋白及血清Scr、BUN和C5b-9水平，升高血Alb水平，减轻足细胞损伤、肾小球基底膜厚度、纤维组织增生、炎性浸润等病理损伤程度；在分子层面，通过免疫组化技术，观察了水蛭素对PHN大鼠肾小球基底膜Nephrin和肾间质FN、Megalin、α-SMA、PAR-1水平及NF-κB和p38 MAPK及其蛋白磷酸化表达水平的影响，发现水蛭素可明显下调PHN大鼠肾间质FN、α-SMA、PAR-1及NF-κB和p38 MAPK蛋白磷酸化的表达水平，上调肾小球基底膜Nephrin和肾间质Megalin水平。通过PCR技术检测，发现水蛭素能够明显下调PHN大鼠肾组织中PAR-1、p38MAPK、NF-κB、MCP-1、MCP-2、MCP-3、FN、ICAM-1、α-SMA及基因表达水平，上调PHN大鼠肾组织中E-Cadherin基因表达水平；且膜性肾病纤维化早期炎症细胞因子MCP-1、MCP-2、MCP-3、ICAM-1、ICAM-3、细胞表面标志物α-SMA及钙粘蛋白E（E-cad）亦有明显下调，促纤维化因子TGF-β1明显上调。以上结果提示了水蛭素对PHN大鼠具有减轻足细胞损伤，改善肾功能，延缓肾间质纤维化进展的作用，其机制可能与水蛭素干预了PHN大鼠特发性膜性肾病发病过程中及其肾间质纤维化早期信号转导通路相关的PAR-1、p38 MAPK、NF-κB等指标表达，进而下调了其下游促炎症指标ICAM-1、ICAM-3、MCP-1、MCP-2、MCP-3、细胞表面标志物a-SMA,E-Cadherin的表达相关。因此，为特发性膜性肾病及其肾间质纤维化治疗提供了数据支持以及新的靶点。

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