“三通针法”电针调控胞浆型磷脂酶A2介导脊髓损伤大鼠炎性反应的研究  

Study of electroacupuncture with“Santong needling method”mediating inflammatory responses by regulating cytosolic phospholipase A2 in spinal cord injury rats

作　　者：姚海华 闵友江 洪冬英 王立 杨宜花 YAO Haihua;MIN Youjiang;HONG Dongying;WANG Li;YANG Yihua(Shanghai Eighth People’s Hospital,Shanghai 200235,China;School of Traditional Chinese Medicine,Nanchang Medical College,Nanchang 300052,China;Shanghai Huangpu District Shunchang Hospital,Shanghai 200025,China)

机构地区：[1]上海市第六人民医院徐汇分院,上海200235 [2]南昌医学院中医学院,南昌300052 [3]上海市黄浦区顺昌医院,上海200025

出　　处：《上海针灸杂志》2023年第7期748-755,共8页Shanghai Journal of Acupuncture and Moxibustion

基　　金：国家自然科学基金项目(81904289,82260963)。

摘　　要：目的探讨电针调控胞浆型磷脂酶A2(cytosolic phospholipase A2,cPLA2)的表达对脊髓损伤(spinal cord injury,SCI)大鼠炎性反应的影响。方法72只清洁级雌性健康SD大鼠制备SCI模型,Basso-Beattie-Bresnahan(BBB)评分后随机分为模型组、电针组、阻断剂组和电针+阻断剂组,另设立假手术组,每组18只。电针组取腰阳关、大椎以及双侧足三里、次髎进行电针治疗;阻断剂组予以花生甙三氟甲基酮(arachidonyl trifluoromethyl ketone,AACOCF3)隔日1次腹腔注射。干预结束后取材,苏木精-伊红(hematoxylin-eosin,HE)染色观察脊髓组织形态学变化,酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)法检测脊髓组织和血清前列腺素E2(prostaglandin E2,PGE2)、血小板活化因子(platelet activating factor,PAF)的含量;Western blot法检测脊髓cPLA2、p-cPLA2蛋白的表达;实时荧光定量PCR法检测脊髓cPLA2 mRNA的表达。结果与假手术组比较,模型组BBB评分明显降低(P<0.01),脊髓组织形态明显受损;脊髓组织和血清PGE2、PAF水平明显升高(P<0.01);脊髓组织p-cPLA2蛋白及cPLA2 mRNA表达水平明显升高(P<0.01)。除电针组cPLA2 mRNA和各治疗组cPLA2蛋白表达水平外,干预后各治疗组大鼠上述指标较模型组都明显逆转(P<0.01)。电针+阻断剂组BBB评分高于电针组和阻断剂组(P<0.01,P<0.05),脊髓组织和血清PGE2、PAF水平低于电针组和阻断剂组(P<0.05),cPLA2 mRNA表达水平低于电针组(P<0.01)。阻断剂组脊髓组织和血清PGE2、PAF水平低于电针组(P<0.01)。结论电针可能通过抑制cPLA2的活性,进而降低PGE2和PAF的水平,减轻神经细胞炎性反应,达到改善SCI大鼠神经功能的作用。Objective To investigate the impact of electroacupuncture on inflammatory responses by regulating cytosolic phospholipase A2(cPLA2)in spinal cord injury(SCI)rats.Method Seventy-two healthy clean female Sprague-Dawley rats were prepared as SCI models and randomized into a model group,an electroacupuncture group,a blocker group,and an electroacupuncture+blocker group after being scored by Basso-Beattie-Bresnahan(BBB),and a sham operation group was also established,with 18 rats in each group.The electroacupuncture group received electroacupuncture intervention by selecting Yaoyangguan(GV3),Dazhui(GV14),and bilateral Zusanli(ST36)and Ciliao(BL32),and the blocker group received intraperitoneal injection of arachidonyl trifluoromethyl ketone(AACOCF3)once every other day.Samples were collected after intervention.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of spinal cord tissues;enzyme-linked immunosorbent assay(ELISA)was adopted to detect the contents of prostaglandin E2(PGE2)and platelet-activating factor(PAF)in the spinal cord and serum;Western blot was used to detect the expression of cPLA2 and p-cPLA2 proteins in the spinal cord;quantitative real-time polymerase chain reaction was employed to determine the expression of cPLA2 mRNA in the spinal cord.Result Compared with the sham operation group,the model group rats had a significant decrease in the BBB score(P<0.01)and obvious morphological damage to spinal cord tissues,the PGE2 and PAF levels increased markedly in the spinal cord tissues and serum(P<0.01),and the expression levels of p-cPLA2 protein and cPLA2 mRNA also increased notably in the spinal cord tissues(P<0.01).After intervention,all the above indexes in all treatment groups went opposite to those in the model group(P<0.01)except the expression of cPLA2 mRNA in the electroacupuncture group and the expression of cPLA2 protein.The BBB score was higher in the electroacupuncture+blocker group than in the electroacupuncture group and the blocker group(P<0.01,P<0.05),and the PGE2

关 键 词：电针 脊髓损伤 胞浆型磷脂酶A2 前列腺素E2 血小板活化因子 大鼠 

分 类 号：R2-03[医药卫生—中医学]