Purmorphamine 对 BV2细胞帕金森病相关基因 Nurr1表达的影响

摘要

目的：探讨Purmorphamine（PM）激活小胶质细胞瘤BV2细胞中Sonic hedgehog（SHH）信号通路对帕金森病（PD）相关基因Nurr1表达的影响。方法体外培养BV2小胶质细胞并分为对照组、脂多糖（LPS）处理组、PM＋LPS处理组以及PM处理组，运用荧光定量 PCR（Q‐PCR）检测经LPS处理后BV2细胞中SHH信号通路Smoothened（Smo）、Gli1及Nurr1基因mRNA 表达情况；PM 激活SHH信号通路后，Q‐PCR检测Nurr1 mRNA含量以及炎性反应因子白细胞介素‐1β（IL‐1β）、肿瘤坏死因子‐α（TNF‐α）的mRNA 表达情况。结果（1）与对照组相比，LPS处理后4 h和24 h时Smo和Gli1 mRNA表达均升高（P＜0.01，P＜0.05）；（2）与对照组相比，PM处理组细胞Smo和Gli1 mRNA表达升高（P＜0.01），LPS组Nurr1、IL‐1β、TNF‐αmRNA表达亦均升高（均 P＜0.01）；而PM＋LPS组Nurr1、IL‐1β、TNF‐αmRNA表达均较LPS处理组下降（均 P＜0.01）。结论PM激活SHH信号通路能够抑制BV2细胞中Nurr1的表达，并能发挥抑制炎性反应作用。%Objective To investigate the effect of Sonic hedgehog (SHH) signaling pathway activated by purmorphamine (PM ) on the expression of Parkinson‐related gene Nurr1 in BV2 microglial cells. Methods The routinely cultured BV2 microglial cells in vitro were divided into the control group , the lipopolysaccharide (LPS) group ,the PM + LPS group and the PM group.Real time quantitative PCR was used to detect contents of SHH relevant gene Smoothened (Smo ) , Gli1 and Nurr1 mRNA in BV2 cell after LPS stimulation. The contents of Nurr1 and the expression of inflammatory factor ‐1β (IL‐1β) , tumor necrosis factor‐α (TNF‐α) mRNA were detected by real time quantitative PCR after SHH signaling pathway activation by PM. Results (1) Compared with the control group , the expression levels of Smo and Gli1 significantly increased 4 hours and 24 hours after LPS stimulation (P<0.01 , P<0.05) . (2) Compared with the control group , the expression levels of Smo and Gli1 significantly increased after PM treatment ( P< 0.01 , respectively ) . The expression levels of Nurr1 and IL‐1β, TNF‐α also significantly increased in the LPS group ( P< 0.01 , respectively ) . Compared with the LPS group , the expression levels of Nurr1 , IL‐1βand TNF‐α decreased in the PM + LPS group (P< 0.01 , respectively). Conclusions SHH signaling pathways activated by PM can inhibit the expression of Nurr1 in BV2 cells , and play a role in inhibiting inflammation.