白藜芦醇预处理减轻神经元焦亡及对神经元缺血性损伤的保护作用

Resveratrol Alleviates the Effect of Neuron Pyroptosis on Ischemic Injury of Neurons

目的探讨白藜芦醇(Res)调控炎性相关蛋白——Nod样受体蛋白3(NLRP3)表达干预神经元焦亡的作用和调控机制。方法培养原代神经元至成熟,随机分为空白组(Sham组)、生理盐水组(Con组)和Res预处理组(Res组)。先对Res组和Con组进行预处理72h,再对3组进行氧糖剥夺损伤6h,最后恢复正常培养液培养2h后取材。利用荧光免疫组化法检测NLRP3的表达情况;利用Western blot检测NLRP3的蛋白水平表达变化。利用标准ELISA试剂盒检测IL-1β和IL-18表达情况。结果与Sham组比较,Con组神经元大量死亡,但存活神经元中NLRP3的染色阳性率高,而Res 10μmol/L、50μmol/L和100μmol/L组均有较多神经元存活,且NLRP3染色阳性率较高;与Con组比较,Res组的NLRP3表达显著降低(均P<0.05),且IL-1β和IL-18表达也明显减少(均P<0.05),各组间差异有统计学意义(P<0.05)。结论 Res预处理下调NLRP3的表达,并减轻IL-1β和IL-18分泌和释放,发挥抗神经元焦亡的作用。

Objective To investigate the effect of resveratrol(Res)on the neuron pyroptosis by regulating the expression of the inflammatory-related protein Nod-like receptor family pyrin domain containing 3(NLRP3)and its mechanism.Methods The primary neurons were cultured and then randomly divided into the blank group(sham group),the physiological saline group(control group)and the Res pretreatment group(Res group).Cells in the control and Res groups were pretreated for 72 h,then subjected to oxygen-glucose deprivation(OGD)for 6 h,and finally re-cultivated in the normal culture medium for another 2 h.The expression of NLRP3 was detected by immunofluorescent staining and Western blotting respectively.ELISA kit was used to detect the expression of IL-1βand IL-18.Results In the control group relative to the sham group,agreater number of neurons were found to die,and the positive rate of NLRP3 staining in surviving neurons was significantly increased.More neurons survived,but the positive rate of NLRP3 staining was still higher in Res groups than in the control group.The expression of NLRP3,IL-1βand IL-18 was significantly decreased in Res groups as compared with that in the control group(P<0.05),and the difference was statistically significant among the 10,50 and 100μmol/L Res groups(P<0.05).Conclusion The Res pretreatment can effectively protect against neuron pyroptosis by down-regulating the expression of NLRP3 and the secretion and release of IL-1βand IL-18.