针刺松弛哮喘急性发作大鼠气道平滑肌的作用机制  

Effect and mechanism of acupuncture on airway smooth muscle relaxation during acute asthma attack in rats

作　　者：乔赟[1] 梁燕娣 卓少元 杨雪捷 梁韬[1] 凌希[1] 张倩[1] 时宜蓉 易蔚[3] QIAO Yun;LIANG Yandi;ZHUO Shaoyuan;YANG Xuejie;LIANG Tao;LING Xi;ZHANG Qian;SHI Yirong;YI Wei(College of Acupuncture-Moxibustion and Tuina,Guangxi University of CM,Nanning 530200,China;Basic Medicine School,Guangxi University of CM,Nanning 530200,China;Graduate School,Guangxi University of CM,Nanning 530200,China)

机构地区：[1]广西中医药大学针灸推拿学院,南宁530200 [2]广西中医药大学基础医学院,南宁530200 [3]广西中医药大学研究生院,南宁530200

出　　处：《中国针灸》2024年第3期295-302,共8页Chinese Acupuncture & Moxibustion

基　　金：2020年广西自然科学基金项目:2020GXNSFAA297021。

摘　　要：目的:探讨针刺“肺俞”“定喘”和“孔最”“鱼际”松弛哮喘急性发作大鼠气道平滑肌的作用机制,并比较两组穴位及联合配伍的疗效。方法:将40只SPF级雄性SD大鼠随机分成空白组、模型组、肺俞定喘组(针刺“肺俞”“定喘”)、孔最鱼际组(针刺“孔最”“鱼际”)和联合组(针刺“肺俞”“定喘”“孔最”“鱼际”),每组8只。除空白组外,其余各组均以卵清蛋白(OVA)联合氢氧化铝凝胶的方法诱导大鼠哮喘急性发作模型。第15天起,除空白组和模型组外,其余各组分别予针刺干预,每次30 min,每日1次,连续14 d。比较各组大鼠的引喘时间,应用肺功能检测仪测定各组大鼠气道阻力(RL)和肺顺应性(Cdyn),ELISA法检测各组大鼠血清及支气管肺泡灌洗液(BALF)内皮素1(ET-1)、肿瘤坏死因子α(TNF-α)、环磷酸腺苷(cAMP)、环磷酸鸟苷(c GMP)含量,Masson染色及透射电镜观察各组大鼠气道平滑肌形态及超微结构,实时荧光定量PCR法检测各组大鼠肺组织ET-1、β_(2)肾上腺素受体(β_(2)-AR)mRNA表达,Western blot法检测各组大鼠肺组织ET-1、β_(2)-AR蛋白表达。结果:与空白组比较,模型组大鼠引喘时间缩短(P<0.05),不同乙酰甲胆碱浓度(0.025 mg/kg、0.05 mg/kg、0.1 mg/kg、0.2 mg/kg)下RL均升高、Cdyn均降低(P<0.05);与模型组比较,肺俞定喘组、孔最鱼际组及联合组大鼠引喘时间延长(P<0.05),不同乙酰甲胆碱浓度下RL均降低、Cdyn均升高(P<0.05);与肺俞定喘组比较,联合组大鼠引喘时间延长(P<0.05)。与空白组比较,模型组大鼠血清及BALF ET-1、TNF-α、cGMP含量升高(P<0.05),cAMP含量降低(P<0.05);与模型组比较,肺俞定喘组、孔最鱼际组、联合组大鼠血清及BALF ET-1、TNF-α、c GMP含量降低(P<0.05),cAMP含量升高(P<0.05)。空白组大鼠肺组织结构正常;模型组肺组织可见大量胶原纤维增生,平滑肌增厚,线粒体明显肿胀,嵴大量断裂、减少;孔最鱼际Objective To explore the effect and mechanism of acupuncture at"Feishu"(BL 13)and"Dingchuan"(EXB 1),and"Kongzui"(LU 6)and"Yuji"(LU 10)for relaxing the airway smooth muscle in the rats during acute asthma attack and compare the effect among the two pairs of acupoints and the acupoints combination.Methods Forty SD male rats with SPF grade were randomly divided into a blank group,a model group,a pair-point A group(acupuncture at"Feishu"[BL 13]and"Dingchuan"[EX-B 1]),a pair-point B group(acupuncture at"Kongzui"[LU 6]and"Yuji"[LU 10])and a point combination group(acupuncture at"Feishu"[BL 13],"Dingchuan"[EX-B 1],"Kongzui"[LU 6]and"Yuji"[LU 10]),with 8 rats in each group.Except the rats in the blank group,the model of acute asthma attack was induced by ovalbumin(OVA)combined with aluminum hydroxide gel in the rest groups.Started on the 15th day of modeling,except in the blank group and the model group,acupuncture was delivered in the other groups,30 min in each intervention,once daily,for 14 days.In each group,the latent period of asthma inducing was measured;the lung resistance(LR)and dynamic lung compliance(Cdyn)were determined using lung function detector;the levels of endothelin-1(ET-1),tumor necrosis factor-α(TNF-α),cyclic adenosine monophosphate(cAMP)and cyclic guanosine monophosphate(cGMP)in serum and bronchoalveolar lavage fluid(BALF)were measured by ELISA;with Masson staining and electron microscopy adopted,the morphology and ultrastructure of airway smooth muscle of the rats were observed;the mRNA and protein expressions of ET-1 and beta-2 adrenergic receptor(β_(2)-AR)were detected by quantitative real-time fluorescence and Western blot,respectively.Results Compared with the blank group,the latent period of asthma inducing was shortened(P<0.05),RL increased and Cdyn decreased(P<0.05)with the different concentrations of methacholine(0.025 mg/kg,0.05 mg/kg,0.1 mg/kg,0.2 mg/kg)in the model group.In the pairpoint A group,the pair-point B group and the point combination group,the latent period of asthma inducin

关 键 词：哮喘 针刺 平滑肌痉挛 内皮素1 β2肾上腺受体 

分 类 号：R245[医药卫生—针灸推拿学]