發(fā)布時間：2018-09-18 10:26

【摘要】：目的： 探討PNPLA3基因I148M過表達Huh7細胞中SREBP-1c與甘油三酯的關(guān)系。為非酒精性脂肪肝的發(fā)病機制研究提供線索。 方法： 用空病毒和攜帶PNPLA3基因I148M突變型的慢病毒感染Huh-7細胞，構(gòu)建空病毒組（對照組）和PNPLA3基因I148M突變型組（突變組）。分別用油紅O染色顯示細胞內(nèi)甘油三酯含量變化，全自動生化分析儀測定各組細胞中甘油三酯（TG）、膽固醇（TC）、谷丙轉(zhuǎn)氨酶（ALT）、谷草轉(zhuǎn)氨酶（AST）含量,熒光定量PCR測定SREBP-1c mRNA的水平變化。Western Blot檢測SREBP-1c蛋白的表達情況。 結(jié)果： 油紅O染色顯示突變組中甘油三酯含量增加。突變組的甘油三酯和總膽固醇含量明顯升高，分別為（0.54±0.03） mmol/L和（0.28±0.03） mmol/L，高于對照組的（0.23±0.02） mmol/L和（0.13±0.02） mmol/L（t值分別為22.58和11.83，，P值均0.01）。谷丙轉(zhuǎn)氨酶和谷草轉(zhuǎn)氨酶也有同樣的表現(xiàn)，分別為（92±10）U/L和（279±12）U/L，明顯高于（170±10）U/L和（585±12）U/L，P值均0.01，差異具有統(tǒng)計學(xué)意義。SREBP-1c mRNA表達量增加，與對照組相比差異具有統(tǒng)計學(xué)意義，P0.01。SREBP-1c蛋白也較對照組有所增加。突變組SREBP-1c與甘油三酯呈正相關(guān)（r=0.912，P0.01），與膽固醇也有一定的相關(guān)性。 結(jié)論： PNPLA3基因I148M多態(tài)性可促進脂肪合成增加，其中甘油三酯的合成增加可能通過SREBP-1c起作用，為探討非酒精性脂肪肝的發(fā)病機制提供了線索。
[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 鹵0.03) mmol/L and (0.28 鹵0.03) mmol/L, respectively, which were significantly higher than those in the control group (0.23 鹵0.02) mmol/L and (0.13 鹵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 鹵10) U / L and (279 鹵12) U / L, respectively, which were significantly higher than those of (170 鹵10) U / L and (585 鹵12) U / L P, respectively. The difference was statistically significant (P < 0.05). SREBP-1c mRNA expression was significantly higher than that of (170 鹵10) U / L and (585 鹵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.
【學(xué)位授予單位】：泰山醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575.5

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