内质网分子伴侣PDIA3在非酒精性脂肪性肝病中的作用及机制研究

发布时间：2018-04-29 17:06

本文选题：非酒精性脂肪性肝病 + 内质网应激　；参考：《浙江大学》2014年硕士论文

【摘要】：目的：非酒精性脂肪性肝病(nonalcoholic fatty liver disease, NAFLD)是近年来影响人民生活健康的常见肝病之一。研究发现,内质网应激(endoplasmic reticulum stress, ERS)在NAFLD发生发展过程中发挥重要作用,其确切分子机制尚未明确。本课题旨在分析内质网分子伴侣蛋白质二硫键异构酶A3前体(protein disulfide isomerase A3precursor, PDIA3/ERp57)在NAFLD中的作用及分子机制。方法：本研究运用软脂酸(palmitic acid, PA)诱导人正常肝细胞系L02细胞建立NAFLD细胞模型。在此基础上,采用小分子RNA干扰(siRNA)、免疫印记、细胞凋亡检测等分子生物学手段,系统研究内质网分子伴侣PDIA3在NAFLD发生发展过程中的作用及分子机制。结果：(1)用PA成功建立NAFLD细胞模型；(2)PDIA3在NAFLD细胞模型中表达上调；(3)NAFLD细胞模型中,siRNA抑制PDIA3表达后引起细胞脂变加重、凋亡增多；(4)NAFLD细胞模型中,siRNA抑制PDIA3表达后引起脂肪酸合成酶(fatty acid synthase, FAS)、磷酸化的PKR样内质网调节激酶(phospho-PKR-like ER kinase, P-PERK)、葡萄糖调节蛋白78(glucose-regulatedprotein78, GRP78)和C/EBP同源蛋白(C/EBP homologous protein, CHOP)表达升高。结论：(1)PDIA3参与NAFLD的发生发展；(2)PDIA3表达抑制通过上调FAS蛋白表达加重软脂酸诱导的肝细胞脂肪变性；(3)PDIA3表达抑制通过激活PERK-CHOP通路加重软脂酸诱导的肝细胞ERS及其相关凋亡。本课题的顺利实施为深入认识NAFLD的分子机制,发现新的诊断和治疗靶标提供了理论依据。
[Abstract]:Objective: nonalcoholic fatty liver disease (NAFLD) is one of the most common liver diseases affecting people's health in recent years. It is found that endoplasmic reticulum stress (ERSs) plays an important role in the development of NAFLD, and its exact molecular mechanism is not clear. The purpose of this study was to analyze the role and molecular mechanism of endoplasmic reticulum molecular chaperone protein disulfide isomerase A3 precursor. PDIA 3 / ERp57 in NAFLD. Methods: NAFLD cell model was induced by palmitic acidin (PAA) in human normal liver cell line L02. On this basis, the role and molecular mechanism of endoplasmic reticulum molecular chaperone PDIA3 in the pathogenesis and development of NAFLD were systematically studied by means of molecular biological methods such as small molecular RNA interference siRNAs, immunological imprinting and cell apoptosis detection. Results (1) NAFLD cell model was successfully established with PA. In NAFLD cell model, the expression of NAFLD was upregulated and increased after inhibiting the expression of PDIA3. The inhibition of PDIA3 expression by siRNA in NAFLD cell model resulted in the increased expression of fatty acid synthase (FASA), phospho-PKR-like kinase (P-PERKN), glucose-regulated protein 78 (GRP78) and C/EBP homologous protein C / EBP homologous protein, CHOP). Conclusion the inhibition of PDIA3 expression in NAFLD by up-regulating the expression of FAS protein increases the inhibition of PDIA3 expression in hepatocytes induced by palmitic acid. The inhibition of PDIA3 expression by activating the PERK-CHOP pathway can aggravate the ERS and its related apoptosis in hepatocytes induced by palmitic acid. The successful implementation of this project provides a theoretical basis for further understanding the molecular mechanism of NAFLD and finding new diagnostic and therapeutic targets.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575

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