艾灸对骶髓损伤后逼尿肌无反射型神经源性膀胱大鼠膀胱组织M2、P2X3受体的影响  

Effect of moxibustion on M2 and P2X3 receptors of bladder tissue in rats with neurogenic bladder of detrusor areflexia after lumbar-sacral spinal cord injury

作　　者：李冰 王永福 任亚锋[2] 冯晓东[2] 白俊敏[2] 牛秋妍 LI Bing;WANG Yong-fu;REN Ya-feng;FENG Xiao-dong;BAI Jun-min;NIU Qiu-yan(School of Rehabilitation Medicine,Henan University of CM,Zhengzhou 450000,China;Rehabilitation Center,First Affiliated Hospital of Henan University of CM,Zhengzhou 450000)

机构地区：[1]河南中医药大学康复医学院,郑州450000 [2]河南中医药大学第一附属医院康复中心,郑州450000

出　　处：《中国针灸》2022年第3期291-297,共7页Chinese Acupuncture & Moxibustion

基　　金：河南省中医药管理局研究基地科研专项项目:2018JDZX010;河南省中医药科学研究专项课题项目:2017JDZX020;河南中医药大学研究生创新创业项目:2018CXCY20。

摘　　要：目的:观察艾灸“关元”“神阙”穴对骶髓损伤后逼尿肌无反射型神经源性膀胱(NB)大鼠膀胱组织乙酰胆碱(Ach)、三磷酸腺苷(ATP)及毒蕈碱型胆碱受体(M)2、嘌呤能受体P2X3的影响,探讨艾灸促进逼尿肌收缩的可能机制。方法:将60只SD大鼠随机分为模型制备组(n=45)和假手术组(n=15),模型制备组采用改良的Hassan Shaker脊髓横断法建立NB模型,假手术组仅咬除椎板暴露骶髓,不进行骶髓横断。从造模成功的大鼠中选取30只随机分为模型组和艾灸组,每组15只。艾灸组大鼠于术后第15天予艾灸“关元”“神阙”穴干预,每穴每次10 min,每日1次,7 d为一疗程,共干预2个疗程。采用尿流动力学检测大鼠膀胱功能,HE染色法观察大鼠膀胱组织形态学改变,生化法测定膀胱组织Ach、ATP含量,Western blot法和实时荧光定量PCR法检测大鼠膀胱组织M2、P2X3受体蛋白和mRNA表达。结果:与假手术组比较,模型组大鼠最大膀胱容量增加、漏尿点压力及膀胱顺应性升高(P<0.05);与模型组比较,艾灸组大鼠最大膀胱容量减小、漏尿点压力及膀胱顺应性降低(P<0.05)。模型组大鼠逼尿肌纤维排列紊乱,膀胱上皮组织断续、细胞排列紊乱、伴大量空泡细胞;艾灸组大鼠逼尿肌纤维较模型组排列清晰,膀胱上皮细胞排列较整齐、空泡细胞减少。与假手术组比较,模型组大鼠膀胱组织Ach、ATP含量减少(P<0.05),M2和P2X3受体蛋白和mRNA表达降低(P<0.05);与模型组比较,艾灸组大鼠膀胱组织Ach、ATP含量增加(P<0.05),M2和P2X3受体蛋白和mRNA表达升高(P<0.05)。结论:艾灸“关元”“神阙”穴可有效改善骶髓损伤后逼尿肌无反射型NB大鼠膀胱功能,其作用机制可能是通过促进Ach释放并上调M2受体表达,进而促进ATP释放、增加P2X3受体表达,以促进逼尿肌收缩。Objective To observe the effect of moxibustion at"Guanyuan"(CV 4)and"Shenque"(CV 8)on acetylcholine(Ach),adenosine triphosphate(ATP)and muscarinic-type choline receptor(M2)and purine receptor P2 X3 in bladder tissue in the rats with neurogenic bladder(NB)of detrusor areflexia after lumbar-sacral spinal cord injury and explore the underlying mechanism of moxibustion for promoting detrusor contraction.Methods Sixty SD rats were randomly divided into a model preparation group(n=45)and a sham-operation group(n=15).In the model preparation group,the modified Hassan Shaker spinal cord transection method was used to prepare the model of NB.In the sham-operation group,the spinal cord transection was not exerted except laminectomy and spinal cord exposure.Among the rats with successfully modeled,30 rats were selected and divided randomly into a model group and a moxibustion group,with 15 rats in each one.On the 15 th day after the operation,moxibustion was applied at"Guanyuan"(CV 4)and"Shenque"(CV 8)in the moxibustion group,10 min at each acupoint,once a day.The consecutive 7-day treatment was as one course and the intervention for 2 courses was required.Urodynamic test was adopted to evaluate bladder function in rats.Using HE staining,the morphological changes in bladder tissue were observed.The content of Ach and ATP in bladder tissue was measured with biochemical method,and the protein and mRNA expression levels of M2 and P2 X3 receptors in bladder tissue were detected with Western blot and real-time fluorescence quantification PCR method.Results Compared with the sham-operation group,the maximum bladder capacity,leakage point pressure and bladder compliance were increased in the rats of the model group(P<0.05).Compared with the model group,the maximum bladder capacity,the leakage point pressure and bladder compliance were decreased in the rats of the moxibustion group(P<0.05).In the model group,the detrusor fibres were arranged irregularly,bladder epithelial tissues were not tightly connected and cell arrangement was

关 键 词：骶髓损伤 神经源性膀胱 逼尿肌无反射 艾灸 M2受体 P2X3受体 

分 类 号：R245.8[医药卫生—针灸推拿学]