JNK/p38 MAPK信号通路在脾虚湿困型溃疡性结肠炎黏膜微环境中的响应及参苓白术散的干预作用

Ulcerative colitis (UC) is a chronic recurrent diseases, which are characterstic of precancerous lesions of colon cancer, and no specific treatment was found untill now. Evidence of pathogenesis of TCM clinical treatment showed that the key pathological mechanisms of UC is syndrome of dampness stagnancy due to spleen deficiency. Based on the theory of excessive dampness causing diarrhea and our previous rearch foundation, this study is to explore dynamic expressions of key molecular of JNK/p38 MAPK signal pathway downstream around JNK, c-myc, c-fos, Caspase-8 and p38MAPK in UC and cross talk of corresponding inflammation medias such as TNF-a,IL-4, IL-6, IL-8 and IL-12 mRNA of UC by using immunohistochemical,western blotting and RT-PCR methods, furtherly, the model rats are treated with Xiangsha Shenlin Baizhu San. Then through correlation analysis, further reveal the mechanism of UC on the basis of JNK/p38 MAPK signal pathway key molecular and the scientific connotations of Shenlin Baizhu San on CAG, and provide theoretical support for clinical application of this therapy and for developing drugs against UC.

溃疡性结肠炎（UC）多呈慢性反复发作,是结肠癌的癌前病变，迄今尚无特异性治疗方法。中医临床证实UC缓解期病机演变多呈脾虚湿困型，故本研究基于"脾虚湿盛则濡泻"和UC病机规律并结合前期基础，拟以脾虚湿困型UC大鼠为研究对象，运用Western blot、免疫组化和RT-PCR技术观察JNK/p38 MAPK信号通路中JNK、c-myc、c-fos、Caspase-8、p38MAPK 基因和蛋白表达水平以及下游关键细胞因子TNF-α、IL-4、IL-6、IL-8、IL-12 mRNA在UC进程中的动态表达规律及其相关性，在此基础上采用参苓白术散进行干预，进一步从跨膜信号转导和炎症因子串联响应角度揭示JNK/p38 MAPK信号通路在脾虚湿困型溃疡性结肠炎黏膜微环境中的响应及参苓白术散的干预作用，并通过相关性分析揭示参苓白术散拮抗UC复发的科学内涵，为临床辨治UC提供有效指导。

溃疡性结肠炎（UC）是目前医学界公认的难治性疾病，已被世界卫生组织列为现代难治病之一。中医学认为，UC缓解期病机多为脾虚湿困，采用参苓白术散加减治疗脾虚湿困型溃疡性结肠炎在临床上取得了良好的效果，但其深层次作用机制尚缺乏深入研究。p38MAPK和JNK是丝裂原活化蛋白激酶（MAPK）家族中两条重要的与炎症相关的信号通路，本实验旨在从JNK/p38 MAPK信号转导通路角度研究UC炎性调控反应机制及参苓白术散对其干预的作用机制。以SPF级Wistar大鼠为实验对象，采用病证结合（即脾虚湿困型UC模型）方法造模成功后，运用ELISA、Western blot、免疫组化和RT-qPCR技术观察参苓白术散干预下大鼠结肠黏膜组织中JNK/p38 MAPK信号通路中JNK、c-myc、c-fos、Caspase-8、p38MAPK等基因、蛋白表达水平以及下游关键细胞因子TNF-α、IL-4、IL-6、IL-8、IL-12 mRNA表达规律及其相关性。结果表明：参苓白术散干预后，运用上述实验方法检测发现IL-4、IL-10表达上调，JNK、c-myc、c-fos、Caspase-8、p38MAPK、TNF-α、IL-6、IL-8、IL-12 、IL-1β表达下调。结论：参苓白术散可通过调控JNK/p38 MAPK信号通路，抑制炎症损伤，减少炎症反应。