槲皮素通过Akt-mTOR途径上调高糖培养RSC96细胞的自噬

Quercetin up-regulates autophagy in RSC96 cells cultured in high glucose via the pathway of Akt-mTOR

目的探讨槲皮素对高糖离体培养的RSC96细胞自噬变化规律的影响。方法 RSC96细胞培养于含有槲皮素的高糖培养基中,超微结构应用透射电子显微镜观察,用免疫荧光法、Western blot法检测Beclin1、LC3Ⅱ、磷酸化的Akt及磷酸化的m TOR蛋白表达,用实时荧光定量PCR法检测Beclin1 mRNA和LC3B mRNA的转录。结果高糖造成RSC96细胞Beclin1和LC3Ⅱ的表达下调,增加Akt和m TOR的磷酸化水平(P<0.05,P<0.01),造成病理形态改变,减少自噬体数量。槲皮素能够上调Beclin1和LC3Ⅱ的表达,下调Akt和m TOR的磷酸化水平(P<0.05,P<0.01),修复病理损伤,增加自噬体数量。结论槲皮素通过下调Akt-m TOR途径磷酸化水平而增加高糖培养RSC96细胞的自噬。

Objective To observe the effect of the quercetin on autophagy of RSC96 cultured in high glucose.Methods RSC96 cells were cultured with high glucose or quercetin. Ultrastructure was performed using transmission electron microscopy. The expression of beclin1,LC3Ⅱ,phosphorylation of Akt（ p-Akt）,and phosphorylation of mT OR（ p-mT OR） were detected by immunofluorescence and Western blot. Real-time fluorescent PCR analysis showed that the level of Beclin1 mRNA and LC3 mRNA. Results Quercetin significantly alleviated the pathological morphology of RSC96 cells and up-regulated the Beclin1 and LC3-Ⅱexpressions（ P〈0. 05,P〈0. 01）,increased autophagosome,but down-regulated the p-Akt and p-mT OR expressions（ P〈0. 05,P〈0. 01）. Conclusions Quercetin up-regulates autophagy in RSC96 cells cultured in high glucose by the pathway of Akt-mT OR.