Injecting RNA interference lentiviruses targeting the muscarinic 3 receptor gene into the bladder wall inhibits neurogenic detrusor overactivity in rats with spinal cord injury

Abstract

Aims To investigate the effects of injecting RNA interference (RNAi) lentiviruses targeting the muscarinic 3 (M3) receptor gene into the bladder wall on bladder activity in rats with spinal cord injury (SCI). Methods Four M3 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 M3 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 M3 RNAi lentivirus with the highest interference efficiency (78.9%) was constructed and identified. Three weeks after injecting M3 RNAi lentiviruses into the bladder wall, Western blotting and qRT‐PCR showed that the M3 receptor was significantly downregulated in the experimental group. Cystometric evaluation suggested that downregulating M3 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 M3 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.

Antifibrosis treatment by inhibition of VEGF, FGF, and PDGF receptors improves bladder wall remodeling and detrusor overactivity in association with modulation of C‐fiber afferent activity in mice with spinal cord injury

“…Previous studies have shown that SCI induces high bladder pressure voiding because of DO and urethral obstruction due to DSD, 10 which is associated with the overexpression of muscarinic receptors and the enhancement of acetylcholine release from parasympathetic nerve terminals. 22 Shang et al 23 revealed that the interference of muscarinic M3 receptors in SCI rats resulted in the improvement of DO. In this study, nintedanib reduced the expression of M2 and M3 receptors although these changes were prominent in the mucosa rather than the detrusor at 4 weeks after SCI in mice.…”

Molecular Mechanism Operating in Animal Models of Neurogenic Detrusor Overactivity: A Systematic Review Focusing on Bladder Dysfunction of Neurogenic Origin

“…The vast majority of the analyzed reports, 74%, use spinal cord injury as an NDO model ( Figure 2 B). From these publications, 59% report complete transection of thoracic segments to induce SCI [ 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 38 , 39 , 43 , 44 , 46 , 48 , 49 , 50 , 55 , 57 , 62 , 63 ], followed by spinal contusion (13%). The use of spinal hemisections was less frequent (10%) [ 42 , 47 , 52 ], as were spinal compressions (8%) [ 32 , 36 , 54 , 56 ] and other SCI methods (10%) [ 41 , 51 , 53 , 58 ] ( Figure 3 ).…”

MicroRNA expression profile in the spinal cord injured rat neurogenic bladder by next-generation sequencing

“…All experimental procedures were implemented in compliance with the National Institute of Health Guidelines for the Care and Use of Laboratory Animals (2) and approved by the Institutional Animal Care and Use Committee of Xuanwu Hospital Capital Medical University (No. 20190128).…”

“…It has been estimated that 250,000 to 500,000 people worldwide are suffering from spinal cord injury and approximately 81% of these patients experience different levels of neurogenic bladder (NB) which is one of the leading causes of their morbidity (1,2). Specifically, detrusor overactivity and detrusor-sphincter dyssynergia are typical NB urodynamic findings, and are diagnosed in 95% and 68% of spinal cord-injured (SCI) patients, respectively (3).…”