基于miR-200b-3p调控Wnt-EMT研究“主客交”理论治疗大鼠肝纤维化的分子机制

The serious impact of liver cirrhosis caused by liver disease generates huge medical and social problems in our motherland.As liver fibrosis plays an inevitable phase in the development of liver cirrhosis, or even liver cancer, naturally， anti-fibrosis research holds key importance in the research fields of preventing liver fibrosis and liver cancer.miR-200 family is an important regulatory factors of epithelium - interstitial transformation (EMT) ， which can reduce β-catenin / Wnt signaling pathway to do a strong inhibition of EMT.Based on the previous research，the high correlationship between miR-200b-3p and treatment of TouxieTongluo (modified Sanjiasan) processed in the anti-fibrosis of rat which under the instructing of Zhukejiao theory discovered by applying high-throughput mircoRNAs sequencing technology，and miR-200b-3p could regulate the Wnt signal pathway which was demonstrated by bio-informatics analysis. Based on the findings above, this topic proposed the hypothesis that treatment of TouxieTongluo (Sanjiasan) instructed by the theory of Zhukejiao could regulate Wnt-EMT pathway to perform anti-fibrosis.In the study, methods as qRT-PCR, Western blot and so on were applied to observe the relationship among miR-200b-3p, Wnt signal pathway and characteristic indexes of EMT, not only to reveal the function of miR-200b-3p regulating Wnt-EMT in anti-fibrosis, but also, initially, to show the molecular mechanism of anti-fibrosis instructed by TCM Zhukejiao theory in the angle of miR-200b-3p regulating Wnt-EMT.

肝纤维化是各种肝病向肝硬化发展的必经阶段，抗肝纤维化研究是有效阻止各种肝病向肝硬化、肝癌发展的关键环节。miR-200家族是一类重要的上皮-间质转化（EMT）调控因子，可通过下调β-catenin /Wnt信号通路抑制EMT。我们前期采用miroRNAs高通量测序技术发现“主客交”理论指导下的透邪通络法（加减三甲散）抗肝纤维化与miR-200b-3p密切相关，生物信息学分析提示miR-200b-3p可调控Wnt信号通路。故我们提出“主客交”理论指导下的透邪通络法通过miR-200b-3p调控Wnt-EMT途径抗肝纤维化的假说,采用定量PCR、免疫印迹等法检测miR-200b-3p、Wnt通路关键信号、EMT特征指标表达，不仅研究miR-200b-3p调控Wnt-EMT在肝纤维化形成中的作用，并首次从“miR-200b-3p调控Wnt-EMT”角度揭示中医“主客交”理论抗肝纤维化的分子机制。

明代著名医家吴又可在《温疫论》中立“主客交”专论，拟“三甲散”名方，后薛生白承其说并加以发展。“主客交”为邪气混处血脉之中，导致络脉瘀滞而成痼疾。此与肝纤维化形成的主要病理环节相似。前期研究证明加减三甲散治疗肝纤维化有良效，本次实验进一步探讨其作用机制。试图从“miR-200b-3p 调控Wnt-EMT”角度揭示“主客交”理论指导下的透邪通络法（加减三甲散）抗肝纤维化的分子机制。从而寻找“主客交”病机理论的实证依据。