益元灸对骶髓损伤后神经源性膀胱大鼠尿流动力学及膀胱组织TRPV4/ATP/P2X5信号通路的影响  

Effect of Yiyuan moxibustion on urodynamics and TRPV4/ATP/P2X5 signal pathway of bladder tissue in rats with neurogenic bladder after sacral cord injury

作　　者：韦慧麟 张芝兰 黄晓萌 陈净 任亚锋[2] 李冰[2] WEI Hui-lin;ZHANG Zhi-lan;HUANG Xiao-meng;CHEN Jing;REN Ya-feng;LI Bing(Henan University of Chinese Medicine,Zhengzhou 450000,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000)

机构地区：[1]河南中医药大学,郑州450000 [2]河南中医药大学第一附属医院,郑州450000

出　　处：《针刺研究》2023年第10期959-968,共10页Acupuncture Research

基　　金：河南省中医药科学研究专项(No.2022JDZX015,20-21ZY2164,2021JDZY022,2018JDZX010)。

摘　　要：目的:观察益元灸对骶髓损伤后逼尿肌无反射型神经源性膀胱(NB)大鼠尿流动力学的作用,探讨益元灸调控膀胱组织中瞬时受体电位香草酸4(TRPV4)、三磷酸腺苷(ATP)、酪氨酸蛋白激酶KIT(C-Kit)和ATP受体P2X5的表达以促进NB大鼠排尿功能恢复的机制。方法:80只雌性SD大鼠,随机选取12只作为假手术组,其余大鼠用骶髓横断法制备膀胱逼尿肌无反射型NB模型,造模成功的大鼠随机分为模型组、益元灸组、益元灸+抑制剂组、抑制剂组,每组12只。术后第15天开始治疗,益元灸组在手法排尿基础上给予益元灸“神阙”“关元”治疗,抑制剂组在手法排尿基础上给予HC067047膀胱灌注(1 mL,1μmol/L,30 min),益元灸+抑制剂组在手法排尿基础上给予益元灸+HC067047膀胱灌注,模型组除手法排尿外不予其他治疗。末次干预结束后采用尿流动力学评价大鼠膀胱功能,HE染色法观察膀胱组织形态,比色法检测膀胱组织中ATP含量,免疫荧光法观察膀胱组织中C-Kit及P2X5的阳性表达率,Western blot法检测膀胱组织中TRPV4、C-Kit和P2X5蛋白表达水平。结果:与假手术组比较,模型组大鼠最大膀胱容量、膀胱顺应性均升高(P<0.01),漏尿点压力、膀胱组织中ATP含量、C-Kit和P2X5的阳性表达率及TRPV4、C-Kit、P2X5蛋白表达水平均降低(P<0.01)。与模型组比较,益元灸组大鼠最大膀胱容量、膀胱顺应性均降低(P<0.01),漏尿点压力、膀胱组织中ATP含量、C-Kit和P2X5的阳性表达率及TRPV4、C-Kit、P2X5蛋白表达水平均升高(P<0.01,P<0.05);抑制剂组大鼠最大膀胱容量、膀胱顺应性均升高(P<0.01),漏尿点压力、膀胱组织中ATP含量、C-Kit和P2X5的阳性表达率及TRPV4、C-Kit、P2X5蛋白表达水平均降低(P<0.05,P<0.01)。与益元灸+抑制剂组比较,益元灸组大鼠最大膀胱容量、膀胱顺应性均降低(P<0.01),漏尿点压力、膀胱组织中ATP含量、C-Kit和P2X5的阳性表达率及TRPV4、C-KiObjectiveTo observe the effect of Yiyuan m oxibustion on urodynam ics and the expressions of transient receptor potential vanilloid 4(TRPV4),adenosine triphosphate(ATP),tyrosine protein kinase KIT(C-Kit)and adenosine triphosphate receptor P2X5 in bladder tissue of rats with detrusor reflex-free neurogenic bladder(NB)after sacral cord injury(SCI),so as to explore its m echanism in prom oting the recovery of urination function of NB rats.MethodsFem ale SD rats were random ly divided into sham operation,m odel,Yiyuan m oxibustion,Yiyuan m oxibus⁃tion+inhibitor(com bination)and inhibitor groups,with 12 rats in each group.The m odel of detruser reflex-free NB after sacral SCI was established by m odified Hassan Shaker spinal cord transection m ethod.The behavioral score of Basso Beasttie Bresnahan(BBB)and urodynam ic indexes were used to evaluate the m odel of rats after operation.Fifteen daysafter m odeling,Yiyuan m oxibustion was applied to“Shenque”(CV8)and“Guanyuan”(CV4)for 20 m in,once daily for 14 days.Rats of the inhibitor and com bination groups were given intravesical instillation of HC067047(1 m L,1μm ol/L,30 m in).After the interventions,urodynam ics was used to evaluate the bladder function of rats.HE staining was used to observe the m orphology of bladder tissue.ATP content in bladder tissue was detected by colorim etric m ethod.The positive expression rates of C-Kit and their receptor P2X5 in bladder tissue were observed by im m unofluo⁃rescence double labeling m ethod,and TRPV4,C-Kit,and P2X5 protein expression levels in bladder tissue were de⁃tected by W estern blot.ResultsCom pared with the sham operation group,the m axim um bladder capacity and blad⁃der com pliance of rats in the m odel group were increased(P<0.01),the leak point pressure,ATP content,the possi⁃tive expression rates of C-Kit and P2X5,and the protein expression levels of TRPV4,C-Kit,P2X5 in bladder tissue were decreased(P<0.01).In com parison with the m odel and com bination groups,the Yiyuan m oxibustion group showed

关 键 词：骶髓损伤 神经源性膀胱 尿流动力学 瞬时受体电位香草酸4 Cajal间质细胞 

分 类 号：R245.8[医药卫生—针灸推拿学]