清肠化湿方基于TNF-α调控MLCK/MLC通路修复溃疡性结肠炎肠黏膜屏障的作用机制研究

Ulcerative colitis is a refractory disease that epithelium mucosa sustained damage as the main pathological changeis. MLCK/MLC signaling pathway participates in the UC mucosal injury.TNF-α can mediate NF-κB to activate MLCK/MLC, increase permeability of the intestinal mucosa, damage the structure and function of intestinal mucosa barrier,and make the immune inflammatory cascade amplification.So it plays an important role in the occurrence and development of ulcerative colitis.Early studies showed that:QingChangHuaShi decoction can significantly improve the clinical symptoms of ulcerative colitis patients,inhibit inflammation, promote ulcer healing,and the mechanism is connected with the regulation of TNF-α、NF-κB expression in intestinal mucosa,whether with MLCK/MLC pathway remains to be elucidated.In this study,we will establish intestinal epithelial cell tight junction injury model and TNBS-induced rat model, compare the decoction with SASP and TNF-α inhibitor，than confirm that QingChangHuaShi decoction can reduce the mucous membrane permeability, repair the intestinal mucosal barrier, reduce inflammation reaction and then paly a role of treatment,because of the machanism that it can inhibite TNF-α to regulate MLCK/MLC pathway.The study can provide objective basis for clinical application of QingChangHuaShi decoctio.

溃疡性结肠炎是以结肠上皮黏膜持续性损伤为主要病理改变的临床难治性疾病，MLCK/MLC通路参与了肠黏膜屏障损伤过程。促炎因子TNF-α介导NF-κB促进MLCK/MLC活化，增加肠黏膜通透性，损伤肠黏膜屏障的结构和功能，继而使免疫炎症级联放大，在溃疡性结肠炎的发生、发展过程中起着重要作用。前期研究表明：临床效方清肠化湿方能够显著改善溃疡性结肠炎患者的临床症状，抑制炎症反应，促进黏膜愈合，其作用机制与调控肠黏膜组织TNF-α、NF-κB表达有关，但与MLCK/MLC通路的关系有待进一步阐明。本研究从体外建立肠上皮细胞紧密连接损伤模型、体内建立TNBS诱导的大鼠结肠炎模型两个方面展开研究，以SASP、TNF-α抑制剂为对照，观察清肠化湿方介导TNF-α调控MLCK/MLC信号通路从而降低黏膜通透性，修复肠粘膜屏障，减轻炎症反应，从而发挥治疗作用的机制，为清肠化湿方的临床应用提供客观依据。

溃疡性结肠炎是以结肠上皮黏膜持续性损伤为主要病理改变的临床难治性疾病，MLCK/MLC通路参与了肠黏膜屏障损伤过程。临床效方清肠化湿方能够显著改善溃疡性结肠炎患者的临床症状，抑制炎症反应，促进黏膜愈合。前期研究表明，其作用机制与调控肠黏膜组织TNF-α、NF-κB表达有关，但与MLCK/MLC通路的关系有待进一步阐明。本研究从体外建立肠上皮细胞紧密连接损伤模型、体内建立TNBS诱导的大鼠结肠炎模型两个方面展开研究，以SASP、TNF-α抑制剂为对照，探讨清肠化湿方促进黏膜愈合机制与调控肠黏膜组织TNF-α、NF-κB以及MLCK/MLC通路的关系，结果发现，促炎因子TNF-α介导NF-κB促进MLCK/MLC活化，增加肠黏膜通透性，损伤肠黏膜屏障的结构和功能，继而使免疫炎症级联放大，在溃疡性结肠炎的发生、发展过程中起着重要作用，清肠化湿方则能介导TNF-α，抑制MLCK/MLC通路活化，从而降低黏膜通透性，促进紧密连接修复，保护肠粘膜屏障，减轻炎症因子的释放，发挥治疗作用。研究结果能够为清肠化湿方的临床应用提供客观依据。