基于肝细胞线粒体PINK1/parkin介导的自噬信号通路的急性肝衰竭调节机制及解毒化瘀颗粒干预效应

The massive necrosis of liver ceslls is the key progression of liver failure.Mitochondria have been serious damaged by oxidative stress,but after injury the mechanism of how fragmented mitochondria degrade is still unknown.The newest clinical and experimental research have show that the Jieduhuayu granules has antioxidant,protecting mitochondria stress effects.So let’s assume that the Jieduhuayu granules can delay the progression of liver failure and revive the liver cells by adjusting PINK1/prakin mediated selective autophagy of liver cell mitochondria.This project is to copy acute liver failure rats model by D-GalN+LPS method,extract liver mitochondria,segregate Cytoplasmic Protein,laser confocal microscope observeing mitochondria protein localization,immunohistochemistry,RT-PCR detecting PINK1/prakin signal pathway and its downstream mitochondrial autophagy specificity protein Drp1、Mfn1、Mfn2、LC3I、LC3II,et al,and by doing this we can finally discuss the of intervention effects of the Jieduhuayu granules in the liver cell mitochondria PINK1/parkin mediate autophagy signaling pathway of liver failure mechanism,and provide provide clinical services of Jieduhuayu granules in provetion of liver failure.

肝细胞大量坏死是肝衰竭病情进展的关键病理环节。线粒体在氧化应激下大量损伤，损伤后的线粒体碎片如何被降解机制未明。最新研究表明：线粒体选择性自噬途径降解受损线粒体是肝细胞存活和再生的关键。前期临床和实验证实解毒化瘀颗粒具有抗氧化应激、保护线粒体作用。我们假说解毒化瘀颗粒可能通过调节PINK1/prakin介导肝细胞线粒体选择性自噬而起到延缓肝衰竭进程、促进肝细胞再生作用。本项目通过D-GalN+LPS复制急性肝衰竭大鼠模型，提取肝脏线粒体，分离胞浆蛋白，激光共聚焦显微镜观察线粒体蛋白定位，电镜观察线粒体自噬体，免疫组化、RT-PCR检测PINK1/prakin介导的自噬信号通路及其下游线粒体自噬特异性蛋白Drp1、Mfn1、Mfn2、LC3I、LC3II等，探讨基于肝细胞线粒体PINK1/parkin介导的自噬信号通路的肝衰竭调节机制及解毒化瘀颗粒干预效应，为解毒化瘀颗粒防治肝衰竭临床服务。

肝细胞大量坏死是肝衰竭病情进展的关键病理环节。线粒体在氧化应激下大量损伤，损伤后的线粒体碎片如何被降解机制未明。最新研究表明：线粒体选择性自噬途径降解受损线粒体是肝细胞存活和再生的关键。前期临床和实验证实解毒化瘀颗粒具有抗氧化应激、保护线粒体作用。我们假说解毒化瘀颗粒可能通过调节PINK1/prakin介导肝细胞线粒体选择性自噬而起到延缓肝衰竭进程、促进肝细胞再生作用。本项目通过D-GalN+LPS复制急性肝衰竭大鼠模型，采用流式细胞仪、TUNEL法检测、WB、 RT-PCR等方法检测PINK1/prakin 介导的自噬信号通路及其下游线粒体自噬特异性蛋白 Drp1、Mfn1、Mfn2、LC3I、LC3II 的表达，揭示了解毒化瘀颗粒具有提高肝细胞合成能力、增强解毒作用降低内毒素产生、减少肝细胞坏死、促进肝再生拮抗肝衰竭的作用；明确调控免疫炎症机制介导PINK1/prakin 自噬信号途径减少线粒体损伤促进肝再生的作用机制；全面阐释解毒化瘀颗粒通过PINK1/prakin 介导肝衰竭肝细胞线粒体自噬的干预效应。已在国内期刊发表相关论文6篇，培养硕士研究生5名，毕业并取得硕士学位3名，在读研究生2名。