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向晶, 卓越, 艾坤, 邓石峰, 许明, 刘琼, 熊明月, 张泓.电针调节P2X7R介导的细胞焦亡途径对骶上脊髓损伤后尿潴留型神经源性膀胱大鼠膀胱排尿功能的影响[J].湖南中医药大学学报,2024,44(8):1391-1400[点击复制] |
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| 电针调节P2X7R介导的细胞焦亡途径对骶上脊髓损伤后尿潴留型神经源性膀胱大鼠膀胱排尿功能的影响 |
| 向晶,卓越,艾坤,邓石峰,许明,刘琼,熊明月,张泓 |
| (湖南中医药大学, 湖南 长沙 410208;永州市中心医院康复医学科, 湖南 永州 425000) |
| 摘要: |
| 目的 观察电针对骶上脊髓损伤后尿潴留型神经源性膀胱大鼠排尿功能的影响，并探讨电针调节嘌呤能离子通道型受体7（purinergic ligand-gated ion channel 7 receptor, P2X7R）介导的细胞焦亡途径在其中的潜在效应机制。方法 从48只雌性SD大鼠中随机抽取12只纳入假手术组，剩余大鼠以T8完全性脊髓横断法建立尿潴留型神经源性膀胱大鼠模型，将已成模的27只大鼠二次随机分为模型组与电针组，每组12只，剩余3只模型大鼠用于实验候补。电针组于术后第19天开始干预，连续10 d，其余两组仅予以捆绑。干预结束后，各组大鼠先行尿流动力学检测，随后快速分离膀胱组织待检，应用HE染色观察膀胱组织形态学变化，透射电镜观察膀胱组织超微结构变化，TUNEL染色检测膀胱组织中细胞损伤情况，ELISA检测膀胱组织中三磷酸腺苷（adenosine triphosphate, ATP）水平，免疫组织化学法和Western blot法检测膀胱组织中P2X7R、NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3, NLRP3）、半胱氨酸天冬氨酸蛋白酶-1（cysteinyl aspartate specific proteinase-1, Caspase-1）、白细胞介素-1β（interleukin-1β, IL-1β）蛋白表达情况。结果 与假手术组比较，模型组大鼠膀胱漏尿点压力、膀胱最大压力、膀胱最大容量显著升高（P＜0.01），以膀胱体积增大伴尿潴留为主要表现；模型组大鼠膀胱组织存在明显的炎性反应且病理改变显著，膀胱组织超微结构可见明显肿胀、变形等细胞损伤，膀胱组织细胞损伤率显著增加（P＜0.01），膀胱组织中ATP含量、P2X7R、NLRP3、Caspase-1、IL-1β的阳性表达及蛋白表达水平均显著升高（P＜0.01）。与模型组比较，电针组大鼠膀胱漏尿点压力、膀胱最大压力、膀胱最大容量降低（P＜0.05），尿潴留症状较轻，膀胱排尿功能改善；电针组大鼠膀胱组织的炎性反应及病理损伤减轻，膀胱组织超微结构变化明显改善，膀胱组织细胞损伤率显著减少（P＜0.01），膀胱组织中ATP含量、P2X7R、NLRP3、Caspase-1、IL-1β的阳性表达及蛋白表达水平均显著降低（P＜0.01）。结论 电针可有效改善骶上脊髓损伤后尿潴留型神经源性膀胱大鼠的膀胱排尿功能，缓解尿潴留症状，减轻膀胱组织病理损伤程度及其炎症反应，其机制与抑制膀胱组织中P2X7R/NLRP3信号通路焦亡蛋白的表达有关。 |
| 关键词: 骶上脊髓损伤 神经源性膀胱 尿潴留 电针 嘌呤能离子通道型受体7 细胞焦亡 |
| DOI：10.3969/j.issn.1674-070X.2024.08.006 |
| 投稿时间：2024-02-08 |
| 基金项目:国家自然科学基金项目（82274666）；湖南省自然科学基金项目（2022JJ30036，2023JJ50425）。 |
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| Effects of electroacupuncture on bladder voiding function in rats with neurogenic bladder featuring urinary retention after suprasacral spinal cord injury through regulation of P2X7R-mediated pyroptosis pathway |
| XIANG Jing, ZHUO Yue, AI Kun, DENG Shifeng, XU Ming, LIU Qiong, XIONG Mingyue, ZHANG Hong |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Department of Rehabilitation Medicine, Yongzhou Central Hospital, Yongzhou, Hunan 425000, China) |
| Abstract: |
| Objective To observe the effects of electroacupuncture on bladder voiding function in rats with neurogenic bladder (NB) featuring urinary retention after suprasacral spinal cord injury (SCI), and to explore the potential mechanisms of electroacupuncture in regulating the pyroptosis pathway mediated by purinergic receptor P2X ligand-gated ion channel 7 (P2X7R). Methods Twelve from 48 female SD rats were randomly selected as the sham-operated group. The remaining rats were used to establish a rat model of NB featuring urinary retention by T8 complete spinal cord transection. Of the 27 successfully modeled rats, 24 were randomized into model group and electroacupuncture group, with 12 rats in each group, and the remaining three model rats were reserved for experimental backups. The electroacupuncture group received intervention on the 19th day after operation for 10 consecutive days, while the other two groups were only restrained. After the intervention, urodynamic tests were first performed on the rats in each group, followed by rapid separation of bladder tissue for examination. HE staining was used to observe the morphological changes in bladder tissue; transmission electron microscopy was used to observe the ultrastructural changes of bladder tissue; TUNEL staining was used to examine cell damage in bladder tissue; ELISA was used to check the level of adenosine triphosphate (ATP) in bladder tissue; immunohistochemistry and Western blot were used to examine the protein expression levels of P2X7R, NOD-like receptor thermal protein domain associated protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (Caspase-1), and interleukin-1β (IL-1β) in bladder tissue. Results Compared with the sham-operated group, the bladder leak point pressure, maximum bladder pressure, and maximum bladder capacity in the model group significantly increased (P＜0.01), with bladder volume increase accompanied by urinary retention as the main manifestations. The model group rats showed significant inflammatory reaction and pathological changes in bladder tissue, with obvious swelling, deformation, and other signs of cell damage in the ultrastructure of bladder tissue, and a significantly increased cell damage rate (P＜0.01). The ATP content, the positive expression and protein expression levels of P2X7R, NLRP3, Caspase-1, and IL-1β in bladder tissue significantly increased (P＜0.01). Compared with the model group, the bladder leak point pressure, maximum bladder pressure, and maximum bladder capacity of the rats in the electroacupuncture group decreased (P＜0.05), with milder symptoms of urinary retention and improved bladder voiding function. The electroacupuncture group showed alleviated inflammatory reaction and pathological damage, significantly improved ultrastructural changes, significantly reduced cell damage rate (P＜0.01), and significantly reduced ATP content, positive expression and protein expression levels of P2X7R, NLRP3, Caspase-1, and IL-1β in the bladder tissue (P＜0.01). Conclusion Electroacupuncture can effectively improve the bladder voiding function in rats with NB featuring urinary retention after suprasacral SCI, alleviate the symptoms of urinary retention, and reduce the degree of pathological damage and inflammatory response in bladder tissue. Its mechanism is related to the inhibition of the expression of pyroptosis protein in P2X7R/NLRP3 signaling pathway in bladder tissue. |
| Key words: suprasacral spinal cord injury neurogenic bladder urinary retention electroacupuncture purinergic receptor P2X ligand-gated ion channel 7 cell pyroptosis |
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