【摘要】：目的明確圍脂滴蛋白2(perilipinin-2,PLIN2)在白藜蘆醇(resveratrol,RSV)改善脂肪變性HepG2細(xì)胞脂滴代謝中的作用機(jī)制。方法 0.2 mmol/L棕櫚酸處理Hep G2細(xì)胞24 h,建立肝細(xì)胞脂肪變性模型。使用0、10、20、40、80μmol/L RSV及轉(zhuǎn)染PLIN2 si RNA或PKI過表達(dá)質(zhì)粒后,再用40μmol/L RSV干預(yù)24 h,油紅O染色檢測脂滴形成情況,酶法檢測細(xì)胞內(nèi)甘油三酯以及細(xì)胞外游離甘油含量,Western blot檢測PLIN2蛋白表達(dá),實時定量PCR檢測PLIN2 m RNA表達(dá)。結(jié)果 RSV可以降低脂肪變性Hep G2細(xì)胞脂滴蓄積程度,促進(jìn)脂滴的脂解。RSV可以通過促進(jìn)磷酸化增強(qiáng)PKA的活性,并呈濃度依賴性地上調(diào)PLIN2 m RNA和蛋白表達(dá)水平(P0.05)。抑制PKA活性或下調(diào)PLIN2表達(dá)后,RSV促進(jìn)脂肪變性Hep G2細(xì)胞脂解的作用顯著減弱(P0.05)。結(jié)論 RSV通過激活PKA-PLIN2通路促進(jìn)Hep G2細(xì)胞脂滴脂解過程,從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.
【作者單位】： 第三軍醫(yī)大學(xué)軍事預(yù)防醫(yī)學(xué)院營養(yǎng)與食品衛(wèi)生學(xué)教研室營養(yǎng)與食品安全研究中心重慶市醫(yī)學(xué)營養(yǎng)研究中心 重慶市營養(yǎng)與食品安全重點(diǎn)實驗室;
【基金】：國家自然科學(xué)基金面上項目(81673154)~~
【分類號】：R575

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