外泌体通过递送miR-130a介导胰腺腺泡细胞-星状细胞间通讯参与胰腺纤维化分子调控

胰腺纤维化是慢性胰腺炎（CP）的主要病理特征，与胰腺星状细胞（PSC）的活化密切相关，但其调节机制仍有待深入研究。本课题组基于前期miRNA芯片、mRNA芯片筛选及生物信息学分析结果提出假说：“被激活的胰腺腺泡细胞释放外泌体，外泌体携载miR-130a， miR-130a递送至受体细胞（PSC）后结合并抑制其靶基因（PPARγ），从而促进PSC活化及胰腺纤维化”。 为验证这一假说，本研究拟通过荧光标记并追踪外泌体及其携载的RNAs在细胞间的转运；通过体外和体内功能获得性研究及功能缺失性研究验证miR-130a对PSC活化及胰腺纤维化等表型的影响；通过双荧光素酶报告实验及RNA结合蛋白免疫沉淀实验(RIP)验证miR-130a是否与PPARγ直接结合。从而阐述外泌体递送的miR-130a介导胰腺腺泡细胞-PSC间通讯的作用机制。本课题的完成，将为CP的发病机制及防治研究提供新的线索。

Pancreatic fibrosis, the main pathological features of chronic pancreatitis (CP), is closely related to the activation of the pancreatitic stellate cell (PSC). But its regulatory mechanism still needs to be further researched. Based on our early experiments about miRNA chip, mRNA chip and bioinformatics analysis, we prompt a hypothesis that “During CP the pancreatic acinar cells can release exosomes in which miR-130a was carried. The carried miR-130a then was delivered to the recipient cell (PSC) and activate PSC targeting PPARγ, finally promote pancreatic fibrosis” . In order to verify the truth of this hypothesis, a tracer technique of fluorescence labelling exosome and the carried RNAs was using to demonstrate the transportation of the exosome and the carried RNAs; in vitro and in vivo study of “gain of function” or “loss of function” was using to demonstrate effects of miR-130a on PSC activation and pancreatic fibrosis; luciferase experiment and RIP (RNA Binding Protein Immunoprecipitation) was using to demonstrate whether the carried miR-130a were combined to it’s target gene -PPARγ.Thereby to demonstrate the function mechanism of communications between pancreatic acinar cells and PSC mediated by exosome and the carried miR-130a. The success of this study will undoubtedly provide solid theoretical basis for the studies of pathogenesis, prevention and treatment of CP.