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。
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
出处
《临床肝胆病杂志》
CAS
2017年第10期1981-1986,共6页
Journal of Clinical Hepatology
基金
陕西省教育厅科研基金(2013JK0788)
关键词
脂肪肝
自噬
脂类代谢
fatty liver
autophagy
lipid metabolism
