PNPLA3基因I148M过表达Huh7细胞中SREBP-1c与甘油三酯的关系

发布时间：2018-09-18 10:26

【摘要】：目的：探讨PNPLA3基因I148M过表达Huh7细胞中SREBP-1c与甘油三酯的关系。为非酒精性脂肪肝的发病机制研究提供线索。方法：用空病毒和携带PNPLA3基因I148M突变型的慢病毒感染Huh-7细胞，构建空病毒组（对照组）和PNPLA3基因I148M突变型组（突变组）。分别用油红O染色显示细胞内甘油三酯含量变化，全自动生化分析仪测定各组细胞中甘油三酯（TG）、胆固醇（TC）、谷丙转氨酶（ALT）、谷草转氨酶（AST）含量,荧光定量PCR测定SREBP-1c mRNA的水平变化。Western Blot检测SREBP-1c蛋白的表达情况。结果：油红O染色显示突变组中甘油三酯含量增加。突变组的甘油三酯和总胆固醇含量明显升高，分别为（0.54±0.03） mmol/L和（0.28±0.03） mmol/L，高于对照组的（0.23±0.02） mmol/L和（0.13±0.02） mmol/L（t值分别为22.58和11.83，，P值均0.01）。谷丙转氨酶和谷草转氨酶也有同样的表现，分别为（92±10）U/L和（279±12）U/L，明显高于（170±10）U/L和（585±12）U/L，P值均0.01，差异具有统计学意义。SREBP-1c mRNA表达量增加，与对照组相比差异具有统计学意义，P0.01。SREBP-1c蛋白也较对照组有所增加。突变组SREBP-1c与甘油三酯呈正相关（r=0.912，P0.01），与胆固醇也有一定的相关性。结论： PNPLA3基因I148M多态性可促进脂肪合成增加，其中甘油三酯的合成增加可能通过SREBP-1c起作用，为探讨非酒精性脂肪肝的发病机制提供了线索。
[Abstract]:Aim: to investigate the relationship between SREBP-1c and triglyceride in Huh7 cells with PNPLA3 gene I148M overexpression. To provide clues for the pathogenesis of non-alcoholic fatty liver. Methods: Huh-7 cells were infected with empty virus and lentivirus carrying I148M mutation of PNPLA3 gene. The empty virus group (control group) and PNPLA3 gene I148M mutant group (mutant group) were constructed. The content of triglyceride in cells was detected by oil red O staining, and the content of triglyceride (TG), cholesterol (TC), transglutaminase (ALT), alanine aminotransferase (AST) was measured by automatic biochemical analyzer. The level of SREBP-1c mRNA was determined by fluorescence quantitative PCR. Western Blot was used to detect the expression of SREBP-1c protein. Results: oil red O staining showed an increase in triglyceride content in mutant group. The contents of triglyceride and total cholesterol in the mutant group were (0.54 卤0.03) mmol/L and (0.28 卤0.03) mmol/L, respectively, which were significantly higher than those in the control group (0.23 卤0.02) mmol/L and (0.13 卤0.02) mmol/L (t = 22.58 and 11.83 mmol/L, respectively). Alanine aminotransferase and alanine aminotransferase also showed the same characteristics, which were (92 卤10) U / L and (279 卤12) U / L, respectively, which were significantly higher than those of (170 卤10) U / L and (585 卤12) U / L P, respectively. The difference was statistically significant (P < 0.05). SREBP-1c mRNA expression was significantly higher than that of (170 卤10) U / L and (585 卤12) U / L, respectively. Compared with the control group, the difference was statistically significant, P0.01. SREBP-1c protein was also increased compared with the control group. SREBP-1c in mutant group was positively correlated with triglyceride (r = 0.912, P 0.01), and was also correlated with cholesterol. Conclusion: the PNPLA3 gene I148M polymorphism can promote the increase of fat synthesis, and the increase of triglyceride synthesis may play a role through SREBP-1c, which provides clues for the pathogenesis of non-alcoholic fatty liver.
【学位授予单位】：泰山医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.5

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