电针调节大鼠三磷酸腺苷酶缓解骶上脊髓损伤后膀胱逼尿肌收缩的研究

关键词：电针  神经源性膀胱  三磷酸腺苷酶  三磷酸腺苷  逼尿肌

Keywords:electroacupuncture  neurogenic bladder  adenosine triphosphatase  adenosine triphosphatase  detrusor muscle

摘要：

      摘要 目的：观察电针对骶上脊髓损伤后神经源性膀胱逼尿肌三磷酸腺苷酶（ATPase）、三磷酸腺苷（ATP）及磷酸化肌球蛋白轻链激酶（p-MLCK）、磷酸化肌球蛋白轻链（p-MLC）水平的影响，探讨电针抑制骶上脊髓损伤后神经源性膀胱逼尿肌反射亢进的作用机制。 方法：40只雌性SD大鼠按随机数字表分为对照组8只，假手术组8只，剩余24只制作尿失禁型神经源性膀胱模型，成模后二次随机分为模型组和电针组，每组8只。术后第19天，电针干预“次髎”、“中极”、“三阴交”，连续干预7d，结束干预行尿流动力学检测，ELISA法测逼尿肌ATPase、ATP、二磷酸腺苷（ADP）、一磷酸腺苷（AMP）含量，Western Blot法测逼尿肌p-MLCK和p-MLC2水平。 结果：模型组较对照组腹腔漏尿点压、膀胱最大压力明显升高（P<0.01），膀胱最大容量明显降低（P<0.01）；逼尿肌ATP含量显著升高（P<0.01），ATPase、ADP和AMP含量显著降低（P<0.01），逼尿肌p-MLC2水平显著升高（P<0.01），p-MLCK水平显著降低（P<0.01）。电针组较模型组膀胱最大压力明显降低（P<0.01），腹腔漏尿点压降低（P<0.05），膀胱最大容量明显升高（P<0.01）；逼尿肌内ATP含量显著降低（P<0.01），ATPase、ADP和AMP含量显著升高（P<0.01），逼尿肌p-MLC2水平明显降低（P<0.01），p-MLCK水平明显升高（P<0.01）。 结论：电针“次髎”、“中极”、“三阴交”穴可有效改善骶上脊髓损伤后逼尿肌亢进大鼠膀胱功能，其作用机制与电针上调逼尿肌中ATPase表达、加速ATP分解，影响p-MLCK、p-MLC2水平，从而抑制逼尿肌收缩有关。

Abstract:

      Abstract Objective： To investigate the impact of electroacupuncture （EA） on adenosine triphosphatase （ATPase）， adenosine triphosphate （ATP）， phosphorylated myosin light chain kinase （p-MLCK）， and phosphorylated myosin light chain （p-MLC） in the detrusor muscle of neurogenic bladder patients who have experienced suprasacral spinal cord injury （SSCI）. The primary objective of this research was to elucidate the mechanism by which EA inhibits detrusor hyperreflexia in these patients. Method： Forty female SD rats were randomly divided into two groups: a control group and a sham operation group， each comprising eight rats. The remaining twenty-four rats were used to create a neurogenic bladder model with incontinence. The rats were then randomly divided into two groups: a model group and an electroacupuncture group， with eight rats in each group. Subsequently， the remaining twenty-four rats were used to create an incontinence-type neurogenic bladder model using a modified T10 spinal cord transection method. The rats were further divided into a model group and an electroacupuncture group， with eight rats in each group. On the nineteenth postoperative day， electroacupuncture was administered at the "Ciliao"， "Zhongji"， and "Sanyinjiao" points for seven consecutive days. Urodynamic tests were conducted after the treatment， and the levels of ATPase， ATP， adenosine diphosphate（ADP）， and adenosine monophosphate（AMP） in the detrusor muscle were measured using enzyme-linked immunosorbent assay（ELISA）. The levels of p-MLCK and p-MLC2 in the detrusor muscle were measured using Western Blot. Result： The model group exhibited higher abdominal leakage point pressure and maximum bladder pressure， as well as lower maximum bladder capacity， compared to the control group（P<0.01）. Additionally， the ATP content in the detrusor muscle was higher， while the levels of ATPase， ADP， and AMP were lower（P<0.01）. The p-MLC2 level in the detrusor muscle was higher， and the p-MLCK level was lower （P<0.01）. In contrast， the electroacupuncture group displayed lower maximum bladder pressure， reduced abdominal leakage point pressure， and increased maximum bladder capacity compared to the model group （P<0.01）. The ATP content in the detrusor muscle was lower， and the levels of ATPase， ADP， and AMP were higher（P<0.01）. Furthermore， the p-MLC2 levels in the detrusor muscle were lower， and the p-MLCK level was higher（P<0.01）. Conclusion： Electroacupuncture at "Ciliao"，"Zhongji"， and "Sanyinjiao" acupoints effectively improves bladder function in rats with detrusor overactivity following SSCI. The underlying mechanism is associated with the upregulation of ATPase expression in the detrusor muscle， promoting ATP hydrolysis and modulating p-MLCK and p-MLC2 levels， thereby inhibiting detrusor muscle contraction.