P2X7R参与炎性刺激大鼠硬脑膜三叉神经节中NLRP3炎性小体的激活

目的采用炎性刺激大鼠硬脑膜疼痛模型,探讨P2X7R是否参与偏头痛发作过程中三叉神经节内NLRP3炎性小体的激活。方法选择雄性SD大鼠42只,免疫印迹实验取27只随机分为9组,每组3只,分别为空白对照组、致炎剂3h组、致炎剂6h组、致炎剂1d组、致炎剂2d组、致炎剂2d对照组、致炎剂3d组、生理盐水对照组和亮蓝G治疗组。免疫荧光双重染色实验取15只随机分为3组,每组5只,分别为空白组、生理盐水组和致炎剂组。观察各组大鼠P2X7R和NLRP3在三叉神经节中的表达情况。结果免疫荧光双重染色结果显示,NLRP3蛋白主要表达于三叉神经元细胞,而P2X7R主要表达于卫星胶质细胞。致炎剂组NLRP3及P2X7R荧光强度明显高于空白组和生理盐水组。免疫印迹结果显示,各致炎剂组NLRP3、P2X7R蛋白表达明显高于空白对照组(P<0.05)。亮蓝G治疗组P2X7R和NLRP3蛋白表达明显低于致炎剂2d对照组和生理盐水对照组(P<0.05)。结论在炎性刺激大鼠硬脑膜模型中可以观察到三叉神经节内P2X7R和NLRP3炎性小体的激活。

P2X7R paiticipates in activation of NLRP3 inflammasome in inflammation-stimulated rat dural trigeminal ganglia

Objective To study whether P2X7Rpaiticipates in activation of NLRP3inflammasome in inflammation-stimulated rat dural trigeminal ganglia.Methods Twenty-seven SD rats undergoing immunoblotting experiments were randomly divided into blank control group,3-h inflammation group,6-h inflammation group,1-day inflammation group,2-day inflammation group,2-day inflammation control group,3-day inflammation group,saline control group,BBG treatment group(3in each group).Fifteen rats undergoing immunofluorescence experiments were randomly divided into blank group,saline group and inflammation group.Expression of P2X7Rand NLRP3protein was detected in trigeminal ganglion of different groups.Results Immunofluorescence double staining showed that the NLRP3was mainly expressed in trigeminal neurons and the P2X7R was chiefly expressed in satellite glial cells.The fluorescence intensity of NLRP3and P2X7Rwas significantly higher in inflammation group than in blank group and saline group.Immunoblotting showed that the expression level of NLRP3and P2X7Rprotein was significantly higher in different inflammation groups than in blank control group.The expression level of P2X7Rand NLRP3 protein was significantly lower in BBG treatment group than in 2-day inflammation contral group and saline control group(P<0.05).Conclusion P2X7Rand NLRP3inflammasome is activated in inflammation-stimulated rat dural trigeminal ganglia models.