电针对局灶脑缺血/再灌注模型大鼠大脑缺血皮质区P2X7R与NLRP3炎性小体表达的影响  

Effects of electroacupuncture on expression of P2X7R and NLRP3 in ischemic cerebral cortex of focal cerebral ischemia and reperfusion rats

作　　者：卞炜 秦文熠[2] Bian Wei;Qin Wenyi(Department of Rehabilitation, Chongqing Yongchuan Traditional Chinese Medicine Hospital, Chongqing, 402160, China;Department of Integrated Traditional Chinese and Western Medicine, The First Affiliated Hospital of Chongqing Medical ,Chongqing, 400016, China)

机构地区：[1]重庆市永川区中医院康复科,重庆402160 [2]重庆医科大学附属第一医院中西医结合科,重庆400016

出　　处：《中国组织化学与细胞化学杂志》2018年第1期23-29,共7页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：目的观察电针治疗对局灶脑缺血/再灌注模型大鼠大脑缺血皮质区嘌呤受体配体门控性离子通道7(purinergic2X7 receptor,P2X7R)和Nod样受体蛋白3(NOD-like receptor pyrin 3,NLRP3)炎性小体表达的影响,探讨电针治疗减轻局灶脑缺血/再灌注炎性损伤的可能机制。方法雄性SD大鼠随机分为假手术组、模型组、电针组,每组16只。采用改良线栓法制备局灶脑缺血/再灌注模型。以大鼠"百会"、"合谷"和"太冲"为电针穴位。采用Bederson行为学评分评价各组大鼠神经功能缺损程度,Western blot和RT-q PCR检测大脑缺血皮区P2X7R、NLRP3蛋白及m RNA表达情况,ELISA检测脑内IL-1β和IL-18含量,荧光共聚焦显微镜检测脑内Iba-1阳性小胶质细胞数量。结果脑缺血再灌注后24h,电针治疗可明显改善神经功能缺损症状。RT-q PCR和Western blot检测显示,模型组大脑皮质缺血区P2X7R和NLRP3 m RNA和蛋白表达较假手术组明显升高,电针治疗可明显减少脑缺血再灌注后P2X7R和NLRP3 m RNA和蛋白表达的升高。ELISA测定表明,与模型组相比,电针组大脑皮质缺血区IL-1β和IL-18含量明显降低。激光扫描共聚焦显微镜观察发现,脑缺血再灌注后24h,电针治疗可明显减少大脑皮质缺血区Iba-1阳性小胶质细胞数量。结论电针可抑制局灶脑缺血/再灌注模型大鼠脑内P2X7R、NLRP3表达的上调,削弱小胶质细胞激活,减轻炎症因子分泌,从而减轻脑缺血/再灌注炎性损伤。Objective To observe the effect of electroacupuncture on the expression of purinergic2X7receptor(P2X7R)and NOD-like receptor pyrin3(NLRP3)in ischemic cerebral cortex and explore the possible mechanism of electroacupuncture in reducing inflammatory injury followed by focal cerebral ischemia/reperfusion.Methods Male SD rats were randomly divided into sham group,focal cerebral ischemia and reperfusion with(I/RE)or without(I/R)electroacupuncture intervention groups,with16rats in each group.The focal cerebral ischemia/reperfusion model was prepared by modified suture method.Electroacupuncture was given at"Baihui","Hegu","Taichong"acupoints.Bederson behavior score was used to evaluate the neurological deficit in each group.Western Blot and RT-qPCR were used to detect the protein and mRNA expression of P2X7R and NLRP3in ischemic cortex.ELISA was used to measure the levels of IL-1βand IL-18.Iba-1positive microglial cells were detected using fluorescence confocal microscopy.Result Twenty-four hours after cerebral ischemia and reperfusion,electroacupuncture treatment significantly improved the symptoms of neurological deficit.The results of RT-qPCR and Western blot showed that the expressions of P2X7R and NLRP3in cerebral cortex were significantly higher in I/R group than in sham group at both mRNA and protein levels.Electroacupuncture significantly reduced such increase.Compared with I/R group,IL-1βand IL-18expression in ischemic area significantly reduced by electroacupuncture.Imaging data indicated that electroacupuncture significantly reduced Iba-1positive microglia in cerebral ischemic cortex compared to non-treatment group24h after reperfusion.Conclusion Electroacupuncture treatment can inhibit ischemia/reperfusion induced upregulation of P2X7R and NLRP3,microglia activation and secretion of inflammatory cytokines,thus alleviating the inflammatory injury in cerebral ischemia and reperfusion.

关 键 词：脑缺血/ 再灌注炎性损伤 电针 嘌呤受体配体门控性离子通道7 NOD 样受体蛋白3 

分 类 号：R332[医药卫生—人体生理学]