Purmorphamine对帕金森病炎症细胞模型的影响及其作用机制

Effects of purmorphamine on inflammatory cell model of parkinson's disease

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 testify the effect of Shh signaling pathway activator Purmorphamine （PM） on PC12cell and the expression changes of inflammatory cytokines, TNF-α, IL-1β, Pelil, in inflammatory cell model of Parkinson＇s disease. MTr method and real-time quantitative PCR （Q-PCR） technology were used to detect cells cytoactive and to determine the expression of related genes, respectively. The results showed ： （ 1 ） MTT analysis revealed that LPS group, PM group and DMSO group were no significant differences in BV2 cell survival which compared with control group （ P 〉 0.05 ） ; but there was a significantly increased survival of PC12cell in the PM ＋ LPS group, which compared with LPS group （P 〈 0.01 ）. （2） Q-PCR analysis showed： there was a significantly increased expression of TNF-α, IL-1β, and Pelil in the LPS-24 h group, and there was a significantly increased expression of Smo and Glil in the LPS-4 h group; but there was a significantly decreased expression of TRAF3 in the LPS-24 h group, compared with the control group （ P 〈 0.01 ）. Analysis of Q-PCR showed that PM-24 h group have a significantly increased expression of Smo and G/il, compared with the control group（P 〈 0.01 ）. Analysis of Q-PCR showed that PM ＋ LPS group have a significantly increased expression of TRAF3, and the expressions of TNF-α, IL-1β, Pelil were significantly decreased compared with the LPS group （P 〈 0.01 ）. The present results suggested that PM increased the survival of PC12 cell, possibly through enhancing the expression of TRAF3.