Background

Ancient prescriptions of Suo Quan Wan (SQW) have therapeutic effects on diabetic bladder dysfunction. However, the underlying mechanism remains unclear. Here, we hypothesized that SQW ameliorates bladder overactivity and regulates neurotransmission via regulating Myosin Va protein expression.

Methods

After diabetic rats were induced by streptozotocin (65 mg/kg), the model of diabetic bladder dysfunction was established by detecting fasting blood glucose, urodynamic test, in vitro muscle strip experiments, and histological examination. One week after induction, SQW was given to observe the therapeutic effect. The expression levels of Myosin Va in control, Model, SQW L and SQW H groups were detected by RT-qPCR, RNAscope and immunofluorescence assay. The expression levels of ChAT, SP, nNOS and VIP proteins were observed by immunofluorescence assay. After knockdown and overexpression of Myosin Va, the expression changes of ChAT, SP, nNOS and VIP and the regulatory role of SQW were observed.

Results

STZ-induced DM rats had significantly higher serum glucose levels and lower body weight. Compared with the diabetic rats, SQW treatment significantly improved urination function with decreased residual volume (RV), bladder compliance (BC), non-voiding contractions (NVCs), and increased voided efficiency (VE). In addition, contractile responses of muscle strips to electrical-field stimulation (EFS), carbachol (CCh), KCl were significantly lower in the SQW H and SQW L groups than those in the model group. RT-qPCR found that the expression of Myosin Va in the bladder tissue or bladder neurons in model group was significantly increased compared with the control group, and SQW treatment significantly decreased the levels of Myosin Va. In DM rats, ChAT and SP expression were significantly increased, while nNOS and VIP expression were significantly decreased, and SQW improved this phenomenon. Interestingly, SQW ameliorated the abnormal expression of ChAT, SP, nNOS and VIP caused by myosin Va knockdown, and Myosin Va overexpression results are consistent with these.

Conclusions

SQW ameliorates overactive bladder and regulate neurotransmission via regulating Myosin Va mRNA and protein expression.

中文翻译：

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锁泉丸通过调节肌球蛋白Va蛋白表达改善膀胱过度活动并调节神经传递

背景

锁泉丸古方对糖尿病膀胱功能障碍有治疗作用。然而，潜在的机制仍不清楚。在这里，我们假设 SQW 通过调节肌球蛋白 Va 蛋白的表达来改善膀胱过度活动并调节神经传递。

方法

链脲佐菌素（65mg/kg）诱导糖尿病大鼠后，通过空腹血糖检测、尿动力学试验、体外肌条实验、组织学检查等方法建立糖尿病膀胱功能障碍模型。诱导1周后给予SQW观察疗效。采用RT-qPCR、RNAscope和免疫荧光法检测对照组、模型组、SQW L组和SQW H组肌球蛋白Va的表达水平。免疫荧光法观察ChAT、SP、nNOS和VIP蛋白的表达水平。Myosin Va敲低过表达后，观察ChAT、SP、nNOS和VIP的表达变化以及SQW的调节作用。

结果

STZ 诱导的 DM 大鼠具有显着更高的血糖水平和更低的体重。与糖尿病大鼠相比，SQW 治疗显着改善了排尿功能，减少了残余体积 (RV)、膀胱顺应性 (BC)、非排尿收缩 (NVC) 和增加排尿效率 (VE)。此外，SQW H 和 SQW L 组肌肉条对电场刺激 (EFS)、卡巴胆碱 (CCh)、KCl 的收缩反应明显低于模型组。RT-qPCR发现模型组膀胱组织或膀胱神经元中肌球蛋白Va的表达较对照组显着升高，SQW治疗显着降低肌球蛋白Va的水平。在DM大鼠中，ChAT和SP表达显着升高。增加，而nNOS和VIP的表达明显下降，而SQW改善了这一现象。有趣的是，SQW改善了肌球蛋白Va敲低引起的ChAT、SP、nNOS和VIP的异常表达，肌球蛋白Va过表达结果与此一致。

结论

SQW 通过调节肌球蛋白 Va mRNA 和蛋白质表达来改善膀胱过度活动症并调节神经传递。