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为探讨Shh信号通路激活剂,2,6,9-三取代嘌呤化合物(Purmorphamine,PM)对帕金森病炎症细胞模型中炎症因子TNF-α,IL-1β和Peli1等的表达,以及对PC12细胞的影响及其作用机制,本研究应用MTT法和实时荧光定量PCR(real-time quantitative PCR,Q-PCR)技术分别检测了细胞活性和相关基因的表达。结果显示:(1)MTT法检测结果显示:与对照组细胞相比,脂多糖(LPS)组、PM组和二甲基亚砜(DMSO)组BV2细胞的存活率无明显差异(P>0.05);而与LPS组相比,PM+LPS组PC12细胞的存活率显著升高(P<0.01)。2)Q-PCR实验结果显示:与对照组相比,LPS-24 h组TNF-α、IL-1β、Peli1的表达显著升高,LPS-4 h组Smo和Gli1的表达显著升高,而LPS-24 h组TRAF3表达显著下降(P<0.01);与对照组相比,PM-24 h组的Smo、Gli1表达显著升高(P<0.01);与LPS组相比,PM+LPS组BV2细胞TRAF3的表达显著升高,而炎症因子TNF-α,IL-1β,Peli1的表达显著下降(P<0.01)。由此我们推测:PM可提高炎症因子作用下PC12细胞的存活率,其作用机制可能与增加TRAF3的表达有关。
To investigate the expression of inflammatory factors TNF-α, IL-1β and Peli1 in the Parkinson’s disease model induced by 2,6,9-trisubstituted purmorphamine (PM) and Shh signaling pathway activator, And its mechanism of action. In this study, MTT assay and real-time quantitative PCR (Q-PCR) were used to detect the cell viability and related gene expression. The results showed that: (1) MTT assay showed that the survival rate of BV2 cells in LPS group, PM group and DMSO group had no significant difference (P> 0.05 ). Compared with LPS group, the survival rate of PC12 cells in PM + LPS group was significantly increased (P <0.01). 2) The results of Q-PCR showed that the expression of TNF-α, IL-1β and Peli1 in LPS-24 h group was significantly increased compared with the control group, while the expressions of Smo and Gli1 in LPS-4 h group were significantly increased Compared with the control group, the expression of Smo and Gli1 in PM-24 h group was significantly increased (P <0.01); Compared with LPS group, the expression of TRAF3 in LPS-24 h group was significantly decreased (P <0.01) BV2 cells TRAF3 expression was significantly increased, while the inflammatory cytokines TNF-α, IL-1β, Peli1 expression was significantly decreased (P <0.01). Thus we speculate: PM can increase the survival rate of PC12 cells under the action of inflammatory cytokines, and its mechanism may be related to increasing the expression of TRAF3.