针刺对脑出血大鼠NLRP3炎性小体的影响  

Effect of acupuncture on NLRP3 inflammatory corpuscle in rats with intracerebral hemorrhage

作　　者：刘昊 张菶 杜嘉 冯培培[1,2] 阮晨 张伟波 朱仲华[1,2] 周驰 李新伟 LIU Hao;ZHANG Beng;DU Jia;FENG Pei-pei;RUAN Chen;ZHANG Wei-bo;ZHU Zhong-hua;ZHOU Chi;LI Xin-wei(Zhejiang Academy of TCM,Hangzhou 310012,China;Department of Acupuncture-Moxibustion,Tongde Hospital of Zhejiang Province,Hangzhou 310012;Department of Acupuncture-Moxibustion,First Afiliated Hospital of Heilongiang University of CM)

机构地区：[1]浙江省中医药研究院,杭州310012 [2]浙江省立同德医院针灸科,杭州310012 [3]黑龙江中医药大学附属第一医院针灸科

出　　处：《中国针灸》2020年第7期757-763,共7页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金青年项目:81804168;黑龙江中医药大学科研基金面上项目:201825。

摘　　要：目的:观察针刺"百会"透"曲鬓"穴对脑出血(ICH)大鼠NLRP3炎性小体的影响,探究针刺促进ICH大鼠神经功能恢复的作用机制。方法:将40只SPF级6周龄雄性SD大鼠随机分为假手术组、模型组、非穴位组、针刺组,每组10只。模型组、非穴位组、针刺组采用自体血注入法制备ICH大鼠模型,假手术组仅行各种手术操作但不注入自体血。造模后约3 h,针刺组予针刺"百会"透患侧"曲鬓"穴,每12小时1次,连续干预7 d;非穴位组取患侧非穴位(平行"百会"穴,距正中线1 cm处)向前进行透刺,余同针刺组操作。干预结束后,评价每组大鼠复合神经功能评分;HE染色法观察大鼠出血半暗带区脑组织形态学变化;免疫组织化学法检测大鼠脑组织Nod样受体蛋白3(NLRP3)炎性小体的表达;Western blot法检测大鼠脑组织NLRP3、白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)蛋白相对表达水平。结果:干预7d后,与假手术组比较,模型组大鼠复合神经功能各分项评分及总评分均明显降低(P<0.01,P<0.05),脑组织神经元水肿、固缩,部分细胞坏死及炎性细胞浸润;与模型组和非穴位组比较,针刺组复合神经功能总评分及肢体对称运动(LS)和触须本体感觉(VP)2个分项评分升高(P<0.01,P<0.05),并能减轻细胞坏死和炎性细胞浸润。与假手术组比较,模型组大鼠脑组织NLRP3炎性小体及蛋白相对表达和IL-1β、IL-18蛋白相对表达增高(P<0.01);与模型组、非穴位组比较,针刺组脑组织NLRP3炎性小体及蛋白相对表达和IL-1β、IL-18蛋白相对表达较低(P<0.01)。结论:针刺"百会"透"曲鬓"穴能下调ICH大鼠脑组织NLRP3、IL-1β、IL-18蛋白表达,抑制炎性反应,进而促进大鼠神经功能恢复。Objective To observe the effect of acupuncture at"Baihui"(GV 20)through"Qubin"(GB 7)on NLRP3 inflammatory corpuscle in rats with intracerebral hemorrhage(ICH),and to explore the action mechanism of acupuncture on promoting the recovery of neural function in rats with ICH.Methods Forty SPF six-week-old male SD rats were randomly divided into a sham operation group,a model group,a non-acupoint group and an acupuncture group,10 rats in each group.The rats in the model group,non-acupoint group and acupuncture group were intervened with autologous blood injection to prepare ICH model,while the rats in the sham operation group were only intervened with operation but not injection with autologous blood.About 3 hours after the establishment of the model,the rats in the acupuncture group were intervened with acupuncture at"Baihui"(GV 20)through"Qubin"(GB 7),once every 12 hours,for 7 days;the rats in the non-acupoint group were intervened with acupuncture at the non-acupoint[parallel to the"Baihui"(GV 20),1 cm next to the midline]on the affected side,and other treatment was the same as the acupuncture group.At the end of the intervention,the composite nerve function score of each group was evaluated;the histomorphology of the hemorrhage penumbra was observed by HE staining;the expression of NLRP3 inflammatory corpuscle in the brain was detected by immunohistochemistry;the relative protein expression levels of NLRP3,interleukin-1β(IL-1β)and interleukin-18(IL-18)in brain were detected by the method of Western blot.Results Seven days into intervention,compared with the sham operation group,each item score and total score of composite nerve function in the model group were significantly reduced(P<0.01,P<0.05).There was edema and karyopyknosis in brain neuron as well as necrocytosis and inflammatory cell infiltration in the model group.Compared with the model group and the non-acupoint group,the total score of composite nerve function and the scores of symmetrical movement of limbs(LS)and proprioception of tentacles(VP)in the

关 键 词：脑出血 针刺 复合神经功能评分 NLRP3炎性小体 白细胞介素-1β(IL-1β) 白细胞介素-18(IL-18) 

分 类 号：R245.97[医药卫生—针灸推拿学]