Injecting RNA interference lentiviruses targeting the muscarinic 3 receptor gene into the bladder wall inhibits neurogenic detrusor overactivity in rats with spinal cord injury
Injecting RNA interference lentiviruses targeting the muscarinic 3 receptor gene into the bladder wall inhibits neurogenic detrusor overactivity in rats with spinal cord injury
复制标题
将针对毒蕈碱3受体基因的RNA干扰慢病毒注射到膀胱壁可抑制脊髓损伤大鼠的神经源性逼尿肌过度活动
DOI：
10.1002/nau.23894
复制
发表时间：
2019-02-01
影响因子：
2
通讯作者：
Ou, Tongwen
中科院分区：
文献类型：
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作者：
Shang, Zhenhua;Jia, Chunsong;Ou, Tongwen
目的探讨膀胱壁注射针对M-3受体基因的RNA干扰（RNAi）慢病毒对脊髓损伤（SCI）大鼠膀胱活动的影响。方法构建4种M-3 RNAi慢病毒，感染原代培养的膀胱平滑肌细胞（BSMCs）。通过Western blotting和定量逆转录聚合酶链反应（qRT-PCR）确定具有最高干扰效率的最佳RNAi慢病毒。雌性Wistar大鼠在T9-10进行脊髓横断，并随机分为3组（n = 8），即空白对照组、阴性对照组和实验组，并在脊髓横断后1周分别向膀胱壁内注射生理盐水、阴性对照shRNA和M-3 RNAi慢病毒。正常大鼠作为正常对照组。观察各组尿动力学指标及膀胱组织形态学变化。结果构建并鉴定了干扰效率最高的M-3 RNAi慢病毒株（78.9%）。膀胱壁注射M-3 RNAi慢病毒3周后，Western blotting和qRT-PCR显示实验组M-3受体表达明显下调。膀胱测压评估表明，下调M-3受体表达可以显著降低SCI大鼠的基础压、残余容量和非排尿收缩次数，增加收缩间期，并显著改善膀胱顺应性。结论膀胱壁注射靶向M-3受体基因的RNAi慢病毒能有效抑制脊髓损伤后神经源性逼尿肌过度活动（NDO）。因此，这种方法可能是一种潜在的治疗脊髓损伤的NDO。
Aims To investigate the effects of injecting RNA interference (RNAi) lentiviruses targeting the muscarinic 3 (M-3) receptor gene into the bladder wall on bladder activity in rats with spinal cord injury (SCI). Methods Four M-3 RNAi lentiviruses were constructed and used to infect primary cultured bladder smooth muscle cells (BSMCs). Western blotting and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were performed to determine the optimal RNAi lentivirus with the highest interference efficiency. Female Wistar rats were subjected spinal cord transection at T9-10 and randomly divided into three groups (n = 8), namely, blank control, negative control, and experimental groups, and injected into the bladder wall with saline, negative control shRNA, and M-3 RNAi lentiviruses, respectively, 1 week after spinal cord transection. The normal rats were used as normal control group. Urodynamic parameters and bladder tissues were evaluated in the different groups. Results An M-3 RNAi lentivirus with the highest interference efficiency (78.9%) was constructed and identified. Three weeks after injecting M-3 RNAi lentiviruses into the bladder wall, Western blotting and qRT-PCR showed that the M-3 receptor was significantly downregulated in the experimental group. Cystometric evaluation suggested that downregulating M-3 receptor expression could substantially decrease basal pressure, residual volume, and non-voiding contraction number, increase intercontraction interval, and significantly improve bladder compliance in rats with SCI. Conclusion Injecting RNAi lentiviruses targeting the M-3 receptor gene into the bladder wall could effectively inhibit neurogenic detrusor overactivity (NDO) due to SCI. Thus, this approach may be a potential treatment for NDO in SCI.