白藜芦醇通过激活PKA-PLIN2通路促进HepG2细胞脂滴脂解

发布时间：2018-11-02 17:14

【摘要】：目的明确围脂滴蛋白2(perilipinin-2,PLIN2)在白藜芦醇(resveratrol,RSV)改善脂肪变性HepG2细胞脂滴代谢中的作用机制。方法 0.2 mmol/L棕榈酸处理Hep G2细胞24 h,建立肝细胞脂肪变性模型。使用0、10、20、40、80μmol/L RSV及转染PLIN2 si RNA或PKI过表达质粒后,再用40μmol/L RSV干预24 h,油红O染色检测脂滴形成情况,酶法检测细胞内甘油三酯以及细胞外游离甘油含量,Western blot检测PLIN2蛋白表达,实时定量PCR检测PLIN2 m RNA表达。结果 RSV可以降低脂肪变性Hep G2细胞脂滴蓄积程度,促进脂滴的脂解。RSV可以通过促进磷酸化增强PKA的活性,并呈浓度依赖性地上调PLIN2 m RNA和蛋白表达水平(P0.05)。抑制PKA活性或下调PLIN2表达后,RSV促进脂肪变性Hep G2细胞脂解的作用显著减弱(P0.05)。结论 RSV通过激活PKA-PLIN2通路促进Hep G2细胞脂滴脂解过程,从而可能改善非酒精性脂肪性肝病。
[Abstract]:Objective to investigate the role of peri lipid drop protein 2 (perilipinin-2,PLIN2) in improving lipid droplet metabolism of HepG2 cells with fatty degeneration by resveratrol (resveratrol,RSV). Methods Hep G2 cells were treated with 0.2 mmol/L palmitic acid for 24 h and the hepatocyte steatosis model was established. After transfection of PLIN2 si RNA or PKI overexpression plasmids, the lipid droplets were detected by oil red O staining, the intracellular triglyceride and extracellular free glycerol content were detected by enzymatic method, and then treated with 40 渭 mol/L RSV for 24 h. The expression of PLIN2 protein was detected by Western blot and the expression of PLIN2 m RNA was detected by real time quantitative PCR. Results RSV could decrease the accumulation of lipid droplets in Hep G2 cells and promote the lipid hydrolysis of Hep G2 cells. RSV could enhance the activity of PKA by promoting phosphorylation, and up-regulate the expression of PLIN2 m RNA and protein in a concentration dependent manner (P0.05). After inhibiting the activity of PKA or down-regulating the expression of PLIN2, the effect of RSV on lipolysis of Hep G2 cells was significantly decreased (P0.05). Conclusion RSV can promote the lipid dripping process of Hep G2 cells by activating the PKA-PLIN2 pathway, which may improve the non-alcoholic fatty liver disease.
【作者单位】：第三军医大学军事预防医学院营养与食品卫生学教研室营养与食品安全研究中心重庆市医学营养研究中心重庆市营养与食品安全重点实验室;
【基金】：国家自然科学基金面上项目(81673154)~~
【分类号】：R575

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