microRNAs在肝星状细胞生物力学响应中的作用及其机制

microRNAs have been found to play essential roles in hepatic stellate cells (HSCs) activation. Hepatic sinusoidal pressure and stretch can directly or indirectly transmit to HSCs and contribute to its activation during portal hypertension. However, little is known concerning the role of microRNAs in HSCs responsing to biomechanics during portal hypertension. In this study, cultured rat HSCs will be exposed to pressure and stretch using a pressure-inducing apparatus and programmed cell traction device Flexercell Strain Unit 4000 respectively. Using the microRNA microarray approach, we will compare the microRNA expression profiles in HSCs following proper pressure or stretch. Then we will screen biomechanics responsive microRNAs compared with previous microRNA expression profiles during HSCs activation in vitro; Next, the recombinant lentivirus encoding biomechanics responsive microRNAs will be constructed and their effects on HSCs responsing to biomechanics will be studied; At last, target gene will be predicted and validated by testing its expression and fluorescence after stable transfection of biomechanics responsive microRNAs in HSCs and fluorescent reporter assays respectively. These researches have important clinical and academic significance to better understanding of pathogenesis of portal hypertension.

microRNAs在肝星状细胞活化过程中有重要的调控作用。肝硬化门脉高压时肝窦压力和肝窦扩张后牵拉力直接或间接地作用于HSC并促进其活化。然而，microRNAs在机械应力诱导HSC活化过程中的调控作用，迄今国内外均尚未见报道。本项目拟应用细胞压力加载和张应变加载装置分别对HSC施加压力和张应变，用microRNA芯片检测力学加载前后HSC的microRNA表达谱，并与自然活化HSC microRNA表达谱比较，筛选力学应答的目标microRNA；构建目标microRNA慢病毒载体，研究目标microRNA在力学因素诱导HSC活化过程中的作用及其机制；预测靶基因，通过检测microRNA转染前后靶基因表达及构建荧光质粒报告系统验证靶基因。这些力学生物学研究对深入了解肝硬化门脉高压的发病机制有重要的理论和临床意义。

microRNAs 在肝星状细胞活化过程中有重要的调控作用。肝硬化门脉高压时肝窦压力和肝窦扩张后牵拉力直接或间接地作用于 HSC 并促进其活化。然而，microRNAs 在机械应力诱导 HSC 活化过程中的调控作用，迄今国内外均尚未见报道。本项目通过应用细胞压力加载和牵拉力加载装置分别对 HSC 施加压力和牵拉力，用 microRNA 芯片检测力学加载前后 HSC 的 microRNA 表达谱，并与自然活化 HSCmicroRNA 表达谱比较，筛选出力学应答敏感的目标microRNA；构建了目标 microRNA的化合物载体，研究力学敏感的目标microRNA 在力学因素诱导 HSC 活化过程中的作用及其机制；并对其进行预测靶基因，通过检测 microRNA 转染前后靶基因的表达及构建荧光质粒报告系统验证力学敏感microRNA靶基因。对深入了解肝硬化门脉高压的发病机制有重要的理论和临床意义。