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

Hui-Lin Wei, Zhi-Lan Zhang, Xiao-Meng Huang, Jing Chen, Ya-Feng Ren, Bing Li
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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 (P<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 (P<0.01). In comparison with the model and combination groups, the Yiyuan moxibustion group showed a decrease in maximum bladder capacity and bladder compliance (P<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 (P<0.01, P<0.05);However, these indicators showed opposite trends in the inhibitor group (P<0.01, P<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.

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益元灸对骶髓损伤后神经源性膀胱大鼠尿动力学及膀胱组织TRPV4/ATP/P2X5信号通路的影响。
目的:观察益元灸对无逼尿肌反射神经源性膀胱(NB)大鼠骶髓损伤(SCI)后尿动力学及膀胱组织瞬时受体电位香草素4(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、益元灸组、益元灸+抑制剂(联合)组和抑制剂组,每组12只。采用改良Hassan-Shaker脊髓横断法建立骶骨SCI后逼尿肌反射自由型NB模型。采用Basso Beasttie Bresnahan(BBB)行为评分和尿动力学指标对大鼠术后模型进行评价。造模后15d,以益元灸“神阙”和“关元”,每次20min,每日1次,共14d。抑制剂组和联合用药组大鼠膀胱内滴注HC067047(1mL,1μmol/L,30min)。干预后,用尿动力学方法评价大鼠膀胱功能。HE染色观察膀胱组织形态。用比色法测定膀胱组织中ATP含量。免疫荧光双标记法观察C-Kit及其受体P2X5在膀胱组织中的阳性表达率,Western印迹法检测膀胱组织中TRPV4、C-Kit和P2X5蛋白的表达水平。结果:与假手术组相比,模型组大鼠的最大膀胱容量和膀胱顺应性增加(PPPPPPP结论:益元灸可改善SCI后膀胱逼尿肌无反射大鼠的尿动力学和膀胱功能,这可能与其激活膀胱组织TRPV4通道,促进膀胱上皮ATP释放,从而增加膀胱Cajal间质细胞及其嘌呤能P2X5受体的表达有关。
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