标本配穴电针预处理抗脑缺血再灌注大鼠炎性损伤的研究

Effect of Electroacupuncture Preconditioning of Combined Biao and Ben Acupoints on Rats of Cerebral Ischemia-Reperfusion Injury from Inflammatory Response

目的从SIRT1/NF-κB炎性通路途径,探讨电针预处理抗脑缺血再灌注损伤的可能作用机制。方法将SD雄性大鼠随机分为假手术组(S组)、缺血组(I/R组)和电针预处理组(EA组),每组各20只。I/R组采用改良的MCAO线栓法缺血2 h后行再灌注3 h,制备大鼠右侧局灶性脑缺血再灌注损伤模型。S组仅分离右侧颈总动脉,不给予其它处理。EA组造模前取"百会""肾俞""三阴交"电针仪预处理,疏密波,频率2/100 Hz,强度1 m A,每15 min为一刺激单元(通电10 min,留针5 min),连续重复4次,共计1 h。各组再灌注3 h后进行神经行为学评分;HE染色法观察海马神经元形态;ELISA法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)等炎性因子浓度;Western-blot方法检测SIRT1和NF-κB蛋白表达。结果电针预处理后,EA组行为学评分、IL-1β、IL-6、TNF-α炎性因子浓度和NF-κB蛋白表达均较模型组降低(P<0.01),SIRT1蛋白表达较模型组升高(P<0.01),神经元形态结构损伤较模型组明显减轻。结论电针预处理可能通过上调内源性脑SIRT1表达,进而抑制NF-κB表达,减轻炎性反应,从而抗脑缺血再灌注损伤。

Objective To research the mechanism of electroacupuncture(EA)preconditioning on cerebral ischemia-reperfusion injury from SIRT1/NF-κB.Methods A total of 60 SPF SD rats were randomly divided to sham group,model group and EA group.The model was set up by the improved MCAO method with ischemia for 2 h and reperfusion for 3 h.Before establishing the model,EA group was electroacupuncture at"Baihui"(GV20),"Shenshu"(SP6)and"Sanyinjiao"(BL23)acupoints and stimulated by instrument of HANS-200 type with 2/100 Hz and 1 m A for 10 min,then resting for 5 min,and this pattern was recycled4 times.After perfusion for 3 h,we observed neuroethology and neurons by HE,and tested the levels of IL-1β,IL-6 and TNF-αby ELISA,and the protein levels of SIRT1 and NF-κB in hippocampus by Western blot.Results After preconditioning of EA,the neurological score,the contents of IL-1β,IL-6 and TNF-αand the protein level of NF-κB in EA group were all obviously lower than those of the model group(P<0.01),but the protein level of SIRT1 in EA group was obviously higher than that of the model group(P<0.01),and the degree of neurons damage in EA group was lighter than that of the model group.Conclusion The preconditioning of EA could resist harm of cerebrall ischemia-reperfusion injury through up-regulating SIRT1 and down-regulating NF-κB to relieve inflammatory reaction.