TLR4/MyD88介导的炎症反应在体外培养癫痫海马组织中的作用研究

Inflammation through TLR4/MyD88 signaling pathway in low-Mg^(2+)-induced epileptic hippocampal slice models

目的 研究在无镁液所致自发性癫痫样放电海马组织中Toll样受体4(TLR4)、髓样分化因子88(MyD88)、IL-1β和IL-6的表达,探讨TLR4介导的炎症反应在癫痫发病中的作用。方法 取新生6～9 d SD鼠的海马,体外培养至7 d时用无Mg^(2+) ACSF灌流3 h,于造模前和造模后1 d、3 d、1 w,采用real-time PCR检测TLR4、MyD88、IL-1β、IL-6基因,采用Western blot检测TLR4和信号蛋白MyD88的表达。结果 与体外培养条件下正常海马组织相比,无镁细胞外液灌流后的癫痫样放电模型的TLR4、MyD88、IL-1β、IL-6 mRNA升高,TLR4和MyD88的蛋白表达升高,并且随时间延长而增强(P <0. 001)。结论 体外培养无Mg^(2+) ACSF所致的癫痫样放电海马组织中,可通过TLR4-MyD88信号通路,激活炎症反应,在癫痫的发病机制中发挥一定的作用。

Objective To explore The expression of toll-like receptor 4(TLR4),medullary differentiation factor(MyD88),IL-1βand IL-6 in low-Mg^2+-induced epileptic hippocampal slice models,and the role of TLR4 mediated inflammatory response in epilepsy was investigated.Methods He hippocampal slice from newborn 6~9 d SD rat was cultured in vitro to 7 days treated by low-Mg^2+ACSF perfusion 3 h.Quantitative real-time-polymerase chain reaction(qRT-PCR)was used for detections of TLR4,MyD88,IL-1βand IL-6 genes,western blot was also determined to detect TLR4 and MyD88 proteins before and after the molding 1 d,3 d,1 w.Results Compared to control group,the mRNA levels of TLR4,MyD88,IL-1βand IL-6 increased significantly in epileptic hippocampal slice after low-Mg^2+ACSF treatment,there was also a significant increase in TLR4,MyD88 proteins expression.Conclusion Neuroinflammation plays an important role in the develpment of epilepsy via TLR4/MyD88 signaling pathway in low-Mg^2+-induced epileptic hippocampal slice models.