针刺“百会”透“曲鬓”对大鼠脑出血后小胶质细胞M1/M2型极化的影响

作者	单位
李丹;朱曦;王中鹏;李龙;万思琦;赵永厚	1.北京中医药大学第三附属医院针灸科（北京 100029）, 2.北京中医药大学东直门医院重点实验室（北京 100700）,3.黑龙江神志医院神志病科（哈尔滨 150036）

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中文摘要:

目的探讨针刺“百会”透“曲鬓”对大鼠脑出血（ICH）后炎性反应程度影响的作用机制。方法 90只SD大鼠按随机数表法分为假手术组、模型组、针刺组、针刺+激动剂阴性对照组、针刺+miR-34a-5p 激动剂组，每组按 1、3、7天3个时间点分为3个亚组，每组6只。针刺+激动剂阴性对照组、针刺+miR-34a-5p 激动剂组于建模前3天侧脑室注射miR-34a-5p 激动剂阴性对照物或miR-34a-5p 激动剂，造模时假手术组颅内注入生理盐水，其余4组采用自体血注入尾壳核制备ICH大鼠模型。建模12 h后针刺干预，针刺“百会”透“曲鬓”。分别采用Western Blot（1、7天）、Real-time PCR（1、3、7天）检测针刺对诱导型一氧化氮合酶（iNOS）、干扰素-γ（IFN-γ）、白细胞介素4（IL-4）、精氨酸酶1（Arg-1）蛋白及mRNA表达的影响。结果在1、7天时，与假手术组同期比较，模型组同期iNOS、IFN-γ、IL-4及Arg-1蛋白表达水平升高（P＜0.01，P＜0.05）；与模型组同期比较，针刺组iNOS、IFN-γ蛋白表达水平降低，Arg-1、IL-4蛋白表达水平升高（P＜0.01）；与针刺+激动剂阴性对照组同期比较，针刺+miR-34a-5p激动剂组iNOS、IFN-γ蛋白表达水平升高，Arg-1、IL-4蛋白表达水平降低（P＜0.01）。在1、3、7天时，与假手术组同期比较，模型组iNOS、IFN-γ、Arg-1、IL-4 mRNA表达水平升高（P＜0.01）；与模型组同期比较，针刺组iNOS、IFN-γ mRNA表达水平降低，Arg-1、IL-4 mRNA表达水平升高（P＜0.01）；与针刺+激动剂阴性对照组同期比较，针刺+ miR-34a-5p激动剂组iNOS、IFN-γmRNA表达水平升高，Arg-1、IL-4 mRNA表达水平降低（P＜0.01）。结论针刺“百会” 透 “曲鬓”减轻大鼠ICH后炎性反应程度影响可能的机制是降低ICH后炎症因子表达，提高抗炎因子表达，抑制小胶质细胞向M1型极化，促进其向M2型极化。

英文摘要:

Objective To explore the effect and mechanism of acupuncture at Baihui（DU20） penetrating Qubin（GB7）on the level of inflammatory response in intracerebral hemorrhage（ICH）model rats. Methods Totally 90 SD rats were divided into the sham-operation group， the ICH group， the acupuncture group， the acupuncture + agomir-NC group， and the acupuncture + miR-34a-5p agomir group by random digit table， each group was subdivided into 3 subgroups according to day 1， 3， and 7，6 in each group. The acupuncture + agomir-NC group and the acupuncture + miR-34a-5p agomir group were given a lateral ventricular injection of either the negative control of the miR-34a-5p activator or the miR-34a-5p activator for 3 days before modeling. The sham-operation group was given an intracranial injection of saline.The ICH rat models were established in all groups except the sham-operation group using intracerebral injecting of autologous blood into the caudate nucleus. Rats were intervened by needling Baihui（DU20）Penetrating Qubin（GB7）12 hours after modeling. The protein and mRNA expression of inducible nitric oxide synthase （iNOS）， interferon-γ（IFN-γ）， interleukin-4（IL-4），and arginase-1 （Arg-1）were detected by Western Blot（day 1 and 7） and Real-time PCR（day 1，3 and 7）. Results At day 1 and 7， compared with the sham-operation group， the expression levels of iNOS， IFN-γ， IL-4， and Arg-1 protein increased in the ICH group （P＜0.01，P＜0.05）. Compared with the ICH group， the expression levels of iNOS and IFN-γ protein reduced， while the expression levels of Arg-1 and IL-4 protein increased in the acupuncture group （P＜0.01）. Compared with the acupuncture + agomir-NC group， the expression levels of iNOS and IFN-γprotein increased， while the expression levels of Arg-1 and IL-4 protein decreased in the acupuncture + miR-34a-5p agomir group（P＜0.01）. At day 1， 3， and 7，compared with the sham-operation group， the expression levels of iNOS， IFN-γ， Arg-1， and IL-4 mRNA increased in the ICH group （P＜0.01）. Compared with the ICH group， the expression levels of iNOS and IFN-γ mRNA reduced， while the expression levels of Arg-1 and IL-4 mRNA increased in the acupuncture group（P＜0.01）. Compared with the acupuncture + agomir-NC group， the expression levels of iNOS and IFN-γ mRNA increased， while the expression levels of Arg-1 and IL-4 mRNA decreased in the acupuncture + miR-34a-5p agomir group （P＜0.01）. Conclusions Baihui（DU20）penetrating Qubin（GB7）could alleviate the degree of inflammatory response in ICH rats. And its mechanism might be related to reducing the expression of inflammatory cytokines， increasing the expression of anti-inflammatory cytokines， inhibiting microglia polarization towards the M1 phenotype， and promoting their polarization towards the M2 phenotype.

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