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

闫蓉; 牛春燕; 于璐; 田宇

doi:10.3969/j.issn.1001-5256.2017.10.027

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

DOI: 10.3969/j.issn.1001-5256.2017.10.027

1. 西安医学院第一附属医院消化内科
2. 河南省人民医院内分泌科
3. 西安医学院第一附属医院检验科

基金项目:

陕西省教育厅科研基金(2013JK0788)；

详细信息

中图分类号: R575.5
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出版历程
- 出版日期: 2017-10-20

Interaction between autophagy and lipid metabolism in cell models of nonalcoholic fatty liver disease

Department of Gastroenterology, The First Affiliated Hospital of Xi' an Medical College

Research funding:

摘要

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...
- 脂肪肝 /
- 自噬 /
- 脂类代谢
Abstract:
Objective To investigate the interaction between autophagy and lipid metabolism in nonalcoholic fatty liver disease ( NAFLD) .Methods Human 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. Results Compared 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) . Conclusion Upregulation of autophagy can promote the elimination of liver fat, while downregulation of autophagy can promote lipid accumulation.
- fatty liver /
- autophagy /
- lipid metabolism

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参考文献(14)

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