非酒精性脂肪性肝病风险基因SIRT3的关联验证和体内过表达效应研究

发布时间：2018-11-02 15:39

【摘要】：目的研究肝脏脂肪酸氧化关键分子SIRT3编码基因变异与非酒精性脂肪性肝病(nonalcoholic fatty liver disease,NAFLD)发生的关系,并在小鼠体内研究人源SIRT3保护性等位型基因过表达对脂肪肝相关表型的影响。方法建立非酒精性脂肪肝的人群研究队列,提取基因组DNA对编码区基因变异rs11246020进行基因分型,并与疾病发病与否做关联分析;建立小鼠NAFLD模型,在小鼠肝脏过表达人源SIRT3基因,8周后检测小鼠肝脏相关生化指标及肝脏HE染色、油红O染色情况,并提取RNA和蛋白采用Real-time PCR和Western blot检测Pparα、Acox1、cpt1a、cpt1b和SIRT3的转录或表达水平。结果与对照组相比,SIRT3基因编码区错意突变rs11246020的GA+AA基因型在NAFLD组分布较低,而GG基因型分布较高,差异具有统计学意义(P0.05);在小鼠肝脏过表达人源SIRT3的A等位型基因后,肝脏脂肪蓄积降低,肝细胞空泡化程度降低,肝脏和血清游离脂肪酸以及肝脏甘油三酯、总胆固醇水平显著降低(P0.05),肝脏脂肪酸代谢相关基因Pparα和Acox1基因转录水平降低(P0.05)。同时,小鼠体质量上升被有效控制(P0.05)。结论 SIRT3基因变异rs11246020与NAFLD风险相关,小鼠体内过表达SIRT3可显著改善脂肪酸代谢和肝脏脂肪蓄积。
[Abstract]:Objective to study the relationship between the mutation of SIRT3 gene encoding key molecule of fatty acid oxidation in liver and the occurrence of (nonalcoholic fatty liver disease,NAFLD in non-alcoholic fatty liver disease. The effects of human SIRT3 protective allele overexpression on fatty liver related phenotypes were studied in mice. Methods A study cohort of non-alcoholic fatty liver was established and genomic DNA was extracted for genotyping of gene variation rs11246020 in the coding region, and correlation analysis was made with the incidence or absence of the disease. The mouse NAFLD model was established. The human SIRT3 gene was overexpressed in the mouse liver. After 8 weeks, the relative biochemical indexes of the liver and the HE staining and the oil red O staining of the liver were detected. The RNA and protein were extracted to detect Ppar 伪 and Acox1,cpt1a, by Real-time PCR and Western blot. The transcription or expression level of cpt1b and SIRT3. Results compared with the control group, the GA AA genotype of rs11246020 was lower in the NAFLD group than that in the control group, but the distribution of GG genotype was higher in the NAFLD group, the difference was statistically significant (P0.05). After over-expression of human SIRT3 A allele in the liver of mice, the accumulation of liver fat decreased, the degree of vacuolation of liver cells decreased, the levels of free fatty acids and triglycerides in liver and serum decreased significantly (P0.05). The transcription levels of Ppar 伪 and Acox1 genes were decreased (P0.05). At the same time, the increase of body mass was effectively controlled (P0.05). Conclusion rs11246020 mutation of SIRT3 gene is associated with the risk of NAFLD. Overexpression of SIRT3 in mice can significantly improve fatty acid metabolism and hepatic fat accumulation.
【作者单位】：第三军医大学军事预防医学院营养与食品卫生学教研室重庆市营养与食品安全重点实验室重庆市医学营养研究中心;
【基金】：国家自然科学基金面上项目(81502807)~~
【分类号】：R575.5

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