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益元艾灸对骶髓损伤后神经源性膀胱大鼠膀胱组织尿动力学及 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.

机构信息

Henan University of Chinese Medicine, Zhengzhou 450000, China.

The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000.

出版信息

Zhen Ci Yan Jiu. 2023 Oct 25;48(10):959-968. doi: 10.13702/j.1000-0607.20221055.

Abstract

OBJECTIVES

To observe the effect of Yiyuan moxibustion on urodynamics 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 mechanism in promoting the recovery of urination function of NB rats.

METHODS

Female SD rats were randomly divided into sham operation, model, Yiyuan moxibustion, Yiyuan moxibustion+inhibitor (combination) and inhibitor groups, with 12 rats in each group. The model of detruser reflex-free NB after sacral SCI was established by modified Hassan Shaker spinal cord transection method. The behavioral score of Basso Beasttie Bresnahan (BBB) and urodynamic indexes were used to evaluate the model of rats after operation. Fifteen days after modeling, Yiyuan moxibustion was applied to "Shenque" (CV8) and "Guanyuan" (CV4) for 20 min, once daily for 14 days. Rats of the inhibitor and combination groups were given intravesical instillation of HC067047 (1 mL, 1 μmol/L, 30 min). After the interventions, urodynamics was used to evaluate the bladder function of rats. HE staining was used to observe the morphology of bladder tissue. ATP content in bladder tissue was detected by colorimetric method. The positive expression rates of C-Kit and their receptor P2X5 in bladder tissue were observed by immunofluorescence double labeling method, and TRPV4, C-Kit, and P2X5 protein expression levels in bladder tissue were detected by Western blot.

RESULTS

Compared with the sham operation group, the maximum bladder capacity and bladder compliance of rats in the model group were increased (<0.01), the leak point pressure, ATP content, the possitive expression rates of C-Kit and P2X5, and the protein expression levels of TRPV4, C-Kit, P2X5 in bladder tissue were decreased (<0.01). In comparison with the model and combination groups, the Yiyuan moxibustion group showed a decrease in maximum bladder capacity and bladder compliance (<0.01), an increase in leakage point pressure, ATP content, the possitive expression rates of C-Kit and P2X5, and TRPV4, C-Kit, and P2X5 protein expression levels (<0.01, <0.05);However, these indicators showed opposite trends in the inhibitor group (<0.01, <0.05).

CONCLUSIONS

Yiyuan moxibustion can improve the urodynamics and bladder function in rats with bladder detrusor nonreflective after SCI, which may be related to its effect in activating the TRPV4 channel in bladder tissue, promoting the release of ATP from bladder epithelium, thus increasing the expression of bladder Cajal interstitial cells and their purinergic P2X5 receptors.

摘要

目的

观察溢元艾灸对骶髓损伤后逼尿肌反射消失性神经源性膀胱(NB)大鼠尿动力学及瞬时受体电位香草酸 4 型(TRPV4)、三磷酸腺苷(ATP)、酪氨酸蛋白激酶 KIT(C-Kit)和三磷酸腺苷受体 P2X5 表达的影响,探讨其促进 NB 大鼠排尿功能恢复的机制。

方法

雌性 SD 大鼠随机分为假手术组、模型组、溢元艾灸组、溢元艾灸+抑制剂(联合)组和抑制剂组,每组 12 只。采用改良 Hassan Shaker 脊髓横断法建立逼尿肌反射消失性 NB 模型。术后采用 Basso-Beasttie Bresnahan(BBB)行为评分和尿动力学指标评价大鼠模型。建模后 15 天,采用溢元艾灸法对“神阙”(CV8)和“关元”(CV4)进行艾灸 20 min,每日 1 次,共 14 天。抑制剂组和联合组大鼠行膀胱内灌注 HC067047(1 mL,1 μmol/L,30 min)。干预后,采用尿动力学评估大鼠膀胱功能。HE 染色观察膀胱组织形态。比色法检测膀胱组织中 ATP 含量。免疫荧光双标法观察膀胱组织中 C-Kit 及其受体 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,模型组大鼠膀胱最大容量和膀胱顺应性增加(P<0.01),漏点压、ATP 含量、C-Kit 和 P2X5 阳性表达率及膀胱组织中 TRPV4、C-Kit、P2X5 蛋白表达水平降低(P<0.01)。与模型组和联合组比较,溢元艾灸组大鼠膀胱最大容量和膀胱顺应性降低(P<0.01),漏点压、ATP 含量、C-Kit 和 P2X5 阳性表达率及膀胱组织中 TRPV4、C-Kit、P2X5 蛋白表达水平升高(P<0.01,P<0.05);抑制剂组上述指标则呈相反变化趋势(P<0.01,P<0.05)。

结论

溢元艾灸可改善 SCI 后逼尿肌反射消失性 NB 大鼠的尿动力学和膀胱功能,其机制可能与激活膀胱组织 TRPV4 通道,促进膀胱上皮细胞释放 ATP,从而增加膀胱 Cajal 间质细胞及其嘌呤能 P2X5 受体表达有关。

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