TM5275对系膜细胞增殖与凋亡的影响及机制的研究

The abnormal proliferation of mesangial cells (MC) caused by the relative lack of apoptosis and the increase of mesangial matrix are the important reasons for the lesion aggravation of Mesangial proliferative glomerulonephritis (MsPGN). Plasminogen activator inhibitor-1 (PAI-1) can inhibit ECM degradation, promote cell proliferation and inhibit apoptosis. As PAI-1 significantly expresses itself in many kinds of acute or chronic kidney damage, it is a potent fibrosis factor for kidney. TM5275 is a novel PAI-1 inhibitor. Our research shows that by promoting the degradation of ECM and the induction of apoptosis of fibroblast, TM5275 can effectively prevent pulmonary fibrosis. Preliminary experiments show that TM5275 can reduce urinary protein and mesangial proliferation on nephritis rats. Thus we speculate the potential prospects that by promoting the degradation of ECM, inhibiting MC proliferation and enhancing apoptosis, TM5275 can have a great effect on the treatment of MsPGN. In this study, by the model of MC fibrosis and proliferation induced by TGF - beta 1, we observe the effect of TM5275 on MC proliferation and apoptosis, and ECM. After the intervention of the animal MsPGN models with TM5275, we observe the pathological changes of the kidney and the effect of TM 5275 on tissue cell apoptosis and the change of ECM. Through these vitro and vivo experiments, we can explore the effect of TM5275 on mesangial cell proliferation and apoptosis and its regulatory mechanism, which is expected to provide evidences for the application of TM5275 to the treatment of chronic kidney diseases.

系膜细胞（MC）相对性凋亡不足导致其异常增殖以及系膜基质增多是系膜增生性肾小球肾炎(MsPGN)进展的重要原因。纤溶酶原激活物抑制剂-1(PAI-1)具有抑制细胞外基质(ECM)降解、促进细胞增生和抑制其凋亡的作用,在各种急慢性肾损害肾脏中显著地表达，是强效的促肾纤维化因子。TM5275是新颖的PAI-1抑制剂，我们的研究表明其通过促进ECM的降解和诱导成纤维细胞的凋亡而抗肺纤维化，预试验表明TM5275可降低肾炎大鼠尿蛋白和系膜增生。由此推测其通过促进ECM的降解、抑制MC增生和促进其凋亡而具有治疗MsPGN的潜在前景。我们将用TGF-β1诱导的MC纤维化增殖模型，观测TM5275对ECM、MC增殖和凋亡的影响。TM5275干预MsPGN动物模型后，观察肾脏病理改变，TM5275对肾组织细胞凋亡和ECM的影响。通过体内外实验为TM5275应用于临床治疗慢性肾脏病提供理论依据。

系膜细胞（MC）相对性凋亡不足导致其异常增殖以及系膜基质增多是系膜增生性肾小球肾炎(MsPGN)进展的重要原因。纤溶酶原激活物抑制剂-1(PAI-1)具有抑制细胞外基质(ECM)降解、促进细胞增生和抑制其凋亡的作用,在各种急慢性肾损害肾脏中显著地表达，是强效的促肾纤维化因子。TM5275是新颖的PAI-1抑制剂，我们的研究表明其通过促进ECM的降解和诱导成纤维细胞的凋亡而抗肺纤维化，预试验表明TM5275可降低肾炎大鼠尿蛋白和系膜增生。由此推测其通过促进ECM的降解、抑制MC增生和促进其凋亡而具有治疗MsPGN的巨大前景。我们将用TGF-β1诱导的MC增殖模型，观测TM5275对MC增殖、凋亡和ECM的影响。TM5275干预抗Thy1.1系膜增生性肾炎大鼠动物模型后，观察肾脏病理改变，TM5275对肾组织细胞凋亡和ECM的影响。通过体内外实验为TM5275应用于临床治疗慢性肾脏病提供理论依据。.体外细胞试验结果：TM5275明显抑制TGF-β1诱导的人系膜细胞增殖、并促进其凋亡(P<0.05); TGF-β1诱导的系膜细胞外基质Fibronectin、Collagen-IV的表达显著增多可被TM5275显著抑制(P<0.05)，TM5275干预后，促凋亡蛋白Bax、caspase-3表达明显增加(P<0.05)，阻止凋亡的蛋白Bcl-2未见明显表达。结论: TM5275可抑制TGF-β1诱导的系膜细胞增殖和细胞外基质的表达，同时诱导促凋亡蛋白的表达，促进系膜细胞凋亡。.抗Thy1.1系膜增生性肾炎大鼠动物实验：M5275干预后，肾炎大鼠异常增高的尿蛋白、血肌酐（Scr）、血尿素氮（BUN）显著下降，肾小球细胞增殖程度、小管间质病理损伤显著减轻，肾组织胶原蛋白沉积也显著下降。另外，异常增殖的肾小球细胞显著凋亡，单核巨噬细胞对肾小球的浸润减少，未见肝功能损害与血常规异常。结论 TM5275能显著降低肾炎大鼠尿蛋白，抑制系膜增生，促进异常增殖的肾小球细胞凋亡，减轻其肾损伤，逆转肾炎大鼠所并发的急性肾损伤，且无肝毒性及骨髓抑制等严重不良反应。其作用机制可能与其抑制系膜细胞增殖、促进其凋亡，同时通过抑制细胞外基质合成或促进其降解，以及抑制单核巨噬细胞对肾组织的侵润损害有关。