抵抗素介导AMPK信号通路对HepG2细胞脂质代谢的影响  

The effect of the recombinant human resistin on lipid metabolism by AMPK pathway in HepG2 cells

作　　者：罗招凡[1] 李芳萍[2] 程桦[2] 

机构地区：[1]中山大学孙逸仙纪念医院检验科,广州市510120 [2]中山大学孙逸仙纪念医院内分泌科,广州市510120

出　　处：《实用医学杂志》2017年第11期1743-1747,共5页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:30570886);教育部留学回国人员科研启动基金资助项目(编号:教外司留[2015]1098号)

摘　　要：目的:探讨抵抗素对Hep G2细胞脂质代谢的影响及其与腺苷酸活化蛋白激酶(AMPK)通路的关系。方法:50 ng/m L抵抗素及0.5 mmol/L的棕榈酸作用Hep G2细胞,利用si RNA技术抑制Hep G2细胞AMPKα2亚基表达,采用实时荧光定量RT-PCR技术检测脂质代谢相关基因的m RNA水平,用蛋白印迹技术检测AMPK蛋白磷酸化(P-AMPK-Thr172)水平及乙酰辅酶A羧化酶(ACC)蛋白磷酸化(P-ACCSer79)水平,尼罗红(Nile Red)染色后激光共聚焦显微镜检测胞内脂质含量。结果:在基础及胰岛素刺激条件下,抵抗素增加ACC2 m RNA的表达水平及胞内脂质含量(P<0.05),降低AMPKα2及肝脂肪酶(HL)m RNA的表达、AMPK及ACC磷酸化水平(P<0.05),对ACC1 m RNA表达无影响(P>0.05)。结论:抵抗素可通过AMPK途径影响脂质代谢:脂肪酸合成增加、抑制三酰甘油分解,导致Hep G2肝细胞胞内脂质积聚。Objective To investigate the effect of rh-resistin on lipid metabolism in HepG2 cells and to elucidate its relation to AMPK pathway. Methods We treated the HepG2 cells with 50 ng/ml rh-resistin and 0.5 mmol/L palmitic acid, used siRNA technique to inhibite α2 subunite expression of AMPK in HepG2 cells and quantitative RT-PCR to detect ACC1, ACC2, and HL mRNA expression levels of related lipid metabolism genes. The P-AMPK-Thr172 of AMPK and P-ACC-Ser79 of ACC were determined by Western blotting. The Lipid accumu- lation in cells was determined by images of Laser Scanning Confocal Microscope after Nile red staining. Results Rh-resistin decreased the AMPKα2, HL mRNA expressions and the phosphorylation level of AMPK and ACC in both basal and insulin-stimulated conditions （P 〈 0.05） , had no influence on ACC2 mRNA expressions （P 〉 0.05） , while it increased ACC2 mRNA expressions and cytoplasmic lipid droplets in the same conditions （P 〈 0.05）. Conclusion Rh-resistin may affect lipid metabolism via AMPK pathway with increase of fatty acid synthesis and inhibition of triglyceride catabolism, which leading to lipid accumulation in HepG2 cells.

关 键 词：抵抗素 游离脂肪酸 脂质 AMPK HEPG2细胞 

分 类 号：R3411[医药卫生—基础医学]