摘要
目的探讨LY294002(LY)抑制磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKt)信号通路后对帕金森病(PD)细胞模型中炎症反应的影响。方法体外培养小鼠小胶质瘤细胞(BV2),分为对照组、脂多糖(LPS)组、LY+LPS组及LY组。LPS刺激BV2细胞来诱导炎症反应,运用Western印迹检测PI3K/AKt信号通路标志物磷酸化(p)-AKt蛋白表达情况;LY抑制PI3K/AKt信号通路后,运用荧光定量PCR (Q-PCR)检测炎症因子白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α的mRNA的表达情况,Western印迹检测诱导型一氧化氮合酶(iNOS)蛋白表达情况。结果与对照组相比,LPS组p-AKt蛋白表达显著下降(P<0.01);与LPS组相比,LY+LPS组p-AKt蛋白下降更明显(P<0.01);与对照组相比,LPS组IL-1β、TNF-αmRNA及iNOS蛋白表达均显著升高(均P<0.01);与LPS组相比,LY+LPS组中IL-1β和TNF-αmRNA及iNOS蛋白表达均显著下降(均P<0.01)。结论 LY抑制PI3K/AKt信号通路对BV2细胞中的LPS诱导的炎症反应有一定的抑制作用。
Objective To investigate the effects of PI3K/AKt signaling pathway inhibited by LY294002 on the inflammatory response in the cell model of Parkinson′s disease (PD). Methods The routinely cultured BV2 microglial in vitro were divided into control, lipopolysaccharide (LPS), LY294002 + LPS and the LY294002 groups. LPS was used to stimulate the BV2 cell to induce inflammatory reaction, Western blot was used to detect the expression of p-AKt protein, which was the marker of PI3K/AKt signal pathway. Inhibition of PI3K/AKt signaling pathway by LY294002, real time quantitative PCR was used to detect the expressions of inflammatory factor IL-1β, TNF-α mRNA and Western blot was used to detect the expression of iNOS protein. Results Compared with control group, the expression of p-AKt protein was significantly decreased after LPS stimulation ( P <0.01), compared with the LPS group, the expression of p-AKt protein was decreased more significantly after treated by LY294002 and LPS( P <0.01). Compared with control group, the expressions of IL-1β, TNF-α mRNA and the expression of p-AKt protein were significantly increased after stimulated by LPS(all P <0.01). Compared with the LPS group, the expressions of IL-1β, TNF-α mRNA and p-AKt protein were decreased after treated by LY294002 and LPS(all P <0.01). Conclusions Inhibition of PI3K/AKt signaling pathway by LY294002 could attenuate inflammatory response in BV2 induced by LPS.
作者
邵帅
洪乐鹏
邓雪华
江梅
丁涛
姜经航
王开秀
SHAO Shuai;HONG Le-Peng;DENG Xue-Hua.(Department of Reproductive Medicine, the Second People Hospital of Jingmen City, Jingmen 448000, Hubei, China)
出处
《中国老年学杂志》
CAS
北大核心
2019年第9期2200-2203,共4页
Chinese Journal of Gerontology
基金
广东省中医药局科研项目(20182080)
广东省科学技术厅资助项目(201301)
湖北省自然科学基金项目(2017CFC808
2018CFC859)
荆门市科技计划项目(2018YFZD013)
