Erbin对TGF-β1诱导胰腺上皮细胞间质转化（EMT）的作用机制研究

Chronic pancreatitis(CP) is a clinical refractory disease with resistance to cure, when it reaches the advanced stage, the patients will present with diabetes, exocrine insufficiency or pancreatic cancer, which will affect their quality of life seriously. As we all know, pancreatic fibrosis is one of the most typical histopathological features, but there are not any effective treatments available for it. Erbin, as a new member of LAP family, has the capability to reverse the effects of TGF-β1 on the process of renal epithelial-to-mesenchyal transition(EMT). Whether erbin has a similar impact on pancreatic fibrosis and EMT is yet unknown. Before this, we have already estasblished animal models of CP induced by DBTC and mastered the skills of purifying and culturing pancreatic stellate cell(PSC) . Our preliminary study confirmed that the expression levels of erbin and TGF-β1 increased in both the serum and pancreatic tissue of CP patients. In this study, we will go further to prove that TGF-β1 could activate quiescent PSC and thus induce EMT by means of RT-PCR, Western Blot, recombinant plasmid and the like. We also want to make sure whether erbin can inhibit pancreatic EMT induced by TGF-β1 and do the research on the signal conducting pathway such as Smad and ERK. In a word, we aim to find out the modifying system of pancreatic fibrosis and the molecular mechanism of EMT, to diversify the pathogenesis of pancreatic fibrosis and to provide a new way for the treatment of CP.

慢性胰腺炎（CP）是一种迁延不愈、临床难治性疾病，晚期可进展为糖尿病、胰腺外分泌功能不全或胰腺癌，严重影响患者生活质量，其主要病理学特征是胰腺纤维化，但目前尚无有效干预措施。Erbin是最近发现的LAP家族重要成员，具有逆转TGF-β1诱导肾上皮细胞间质转化（EMT）作用，但对胰腺纤维化和EMT作用尚不清楚。我们前期研究成功构建了二丁基二氯化锡诱导CP纤维化动物模型并分离和原代培养了胰腺星状细胞（PSC），且发现CP患者血清及组织中erbin和TGF-β1显著上调。本研究拟通过RT-PCR、Western Blot和质粒转染等技术，证实TGF-β1可诱导PSC发生EMT，并明确erbin对TGF-β1诱导PSC发生EMT的抑制作用，同时检测Smad和ERK信号转导通路相关基因和蛋白表达，明确其调控胰腺纤维化及EMT的分子机制，丰富胰腺纤维化机制研究，为抑制或逆转胰腺纤维化提供新的治疗靶点。

慢性胰腺炎（CP）以胰腺组织进行性不可逆性损伤为特征，最终导致胰腺内外分泌功能不全，临床常表现为慢性疼痛、消化不良和糖尿病。CP病程长，病情反复，预后不良，目前治疗仅以缓解症状为目的。本研究拟探讨①组织块法分离培养小鼠胰腺星状细胞（PSC），②TGF-β1诱导PSC发生EMT样改变，③Erbin可通过TGF-β信号通路抑制PSC发生EMT改变，进而抑制或逆转胰腺纤维化，④探讨CP发病新机制。结果显示：①组织块法可成功分选CP模型组和对照组PSC，且两组贴壁早期细胞均为静息状态；②PSC体内、外激活后，Erbin和TGF-β1表达均上调；③PSC活化过程发生EMT样改变，E-cadherin表达下降，α-SMA过表达，且上调Erbin、TGF-β/Smad通路及ERK通路靶点分子表达，该过程参与胰腺纤维化的形成；④基因测序发现754个有统计学意义的相关基因；⑤蛋白液相芯片发现OPN和MCP在CP病人中高表达。通过上述研究，期望丰富胰腺纤维化的发生机制并为抑制或逆转胰腺纤维化提供新的治疗靶点，以有效改善CP患者的预后。