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非酒精性脂肪性肝病细胞模型中自噬与脂质代谢的相互调节
Interaction between autophagy and lipid metabolism in cell models of nonalcoholic fatty liver disease
文章发布日期:2017年09月07日  来源:  作者:闫蓉, 牛春燕, 于璐, 等  点击次数:100次  下载次数:9次

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【摘要】:目的探讨非酒精性脂肪性肝病(NAFLD)中自噬与脂质代谢的相互作用。方法体外人肝细胞培养(脂肪变性)制备NAFLD细胞模型,雷帕霉素诱导细胞自噬,3-甲基腺嘌呤抑制细胞自噬,MTT比色法测定细胞活力,ELISA法检测各组细胞TG、ALT、AST、LDH、GGT、Alb水平,IF法检测 LC3-Ⅱ的定位与分布,Western Blot 检测LC3-Ⅱ/LC3-Ⅰ比值。计量资料多组间比较采用方差分析,进一步两两比较采用SNK-q检验。结果诱导自噬组的吸光度值和细胞活率与脂肪变组比较,明显下降(HL-7702细胞q值分别为4.160、4.110,SK-HEP-1细胞q值分别为4.407、4.032;P值均<0.05)。脂肪变组TG、ALT、AST、LDH、GGT、Alb水平与对照组相比,明显升高(HL-7702细胞q值分别为5.316、3.730、4.013、6.967、6.192、5.531,SK-HEP-1细胞q值分别为4.963、3.603、4.774、7.479、6.319、5.193;P值均<0.05)。诱导自噬组TG、ALT、AST、LDH、GGT、Alb水平与脂肪变组相比,明显降低(HL-7702细胞q值分别为4.978、3.695、3.960、5.130、4.695、3.192,SK-HEP-1细胞q值分别为3.846、5575、4184、5.019、4203、3.049;P值均<0.05)。LC3-Ⅱ在各组肝细胞中的标记值,诱导自噬组最高(HL-7702细胞为90.1%,SK-HEP-1细胞为800%),其次是脂肪变组(HL-7702细胞为47.2%,SK-HEP-1细胞为48.4%)及抑制自噬组(HL-7702细胞为30.2%,SK-HEP-1细胞为455%)。诱导自噬组LC3-Ⅱ/LC3-Ⅰ比值与脂肪变组相比,明显升高(HL-7702细胞q值为6786,SK-HEP-1细胞q值为5.926;P值均<0.05)。结论自噬的上调有利于促进肝脏脂肪的清除,而下调则促进脂质的聚积。
【Abstract】:ObjectiveTo investigate the interaction between autophagy and lipid metabolism in nonalcoholic fatty liver disease (NAFLD). MethodsHuman hepatocytes (steatosis) were cultured in vitro to establish a cell model of NAFLD. Rapamycin was used to induce autophagy and 3-methyladenine was used to inhibit autophagy. MTT colorimetry was used to measure cell viability. ELISA was used to measure the levels of triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), gamma-glutamyl transpeptidase (GGT), and albumin (Alb). IF method was used to determine the location and distribution of LC3-II. Western blot was used to measure LC3-II/LC3-I ratio. An analysis of variance was used for comparison of continuous data between groups, and the SNK-q test was used for further comparison between two groups. ResultsCompared with the steatosis group, the induced autophagy group had significant reductions in absorbance and cell viability (HL-7702 cells: q=4.160 and 4.110, P<0.05; SK-HEP-1 cells: q=4.407 and 4.032, P<0.05). Compared with the control group, the steatosis group had significant increases in the levels of TG, ALT, AST, LDH, GGT, and Alb (HL-7702 cells: q=5.316, 3.730, 4.013, 6.967, 6.192, and 5.531, P<0.05; SK-HEP-1 cells: q=4.963, 3.603, 4.774, 7.479, 6.319, and 5.193, P<0.05). Compared with the steatosis group, the induced autophagy group had significant reductions in the levels of TG, ALT, AST, LDH, GGT, and Alb (HL-7702 cells: q=4.978, 3.695, 3.960, 5.130, 4695, and 3.192, P<0.05; SK-HEP-1 cells: q=3.846, 5.575, 4.184, 5.019, 4.203, 3.049, P<0.05). The induced autophagy group had the highest percentage of LC3-II-positive HL-7702 cells (90.1%) and LC3-II-positive SK-HEP-1 cells (80.0%), followed by the steatosis group (47.2% LC3-II-positive HL-7702 cells and 48.4% LC3-II-positive SK-HEP-1 cells) and the autophagy inhibition group (30.2% LC3-II-positive HL-7702 cells and 45.5% LC3-II-positive SK-HEP-1 cells). The induced autophagy group had a significant increase in LC3-II/IC3-I ratio compared with the steatosis group (HL-7702 cells: q=6.786, P<0.05; SK-HEP-1 cells: q=5.926, P<0.05). ConclusionUpregulation of autophagy can promote the elimination of liver fat, while downregulation of autophagy can promote lipid accumulation.
【关键字】:脂肪肝; 自噬; 脂类代谢
【Key words】:fatty liver; autophagy; lipid metabolism
【引证本文】:闫蓉, 牛春燕, 于璐, 等. 非酒精性脂肪性肝病细胞模型中自噬与脂质代谢的相互调节[J]. 临床肝胆病杂志, 2017, 33(10): 1981-1986.

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