| 摘要: |
| 目的 探讨电针关元穴经miRNA-199a-3p调控酪氨酸激酶受体c-kit/ERK通路对神经源性尿潴留大鼠的治疗作用。方法 采用改良脊髓横断法建立神经源性尿潴留大鼠模型60只，按随机数字表法平均分为5组：模型组、电针组、miRNA-199a-3p抑制剂组、miRNA-199a-3p抑制剂阴性对照组、电针+miRNA-199a-3p抑制剂组，另选取SD大鼠12只作为假手术组。使用药物分组干预处理后，检测大鼠尿动力学，比较各组大鼠膀胱最大容量、膀胱基础压、漏尿点压力；采用肌条实验检测大鼠逼尿肌兴奋性，比较各组肌条自发性收缩频率；采用H-E染色观察各组大鼠膀胱组织病理形态改变；用试剂盒测定各组大鼠血清炎症细胞因子环氧化酶2（COX-2）、IL-18、诱导型一氧化氮合酶（iNOS）水平；采用qPCR检测各组大鼠膀胱组织miRNA-199a-3p与c-kit mRNA表达水平；采用蛋白质印迹法检测各组大鼠膀胱组织c-kit/ERK通路蛋白表达水平。结果 与假手术组相比，模型组大鼠膀胱组织呈现严重病理损伤改变，膀胱最大容量、膀胱基础压、漏尿点压力及血清COX-2、IL-18、iNOS水平均升高（P均<0.05），肌条自发性收缩频率、膀胱组织miRNA-199a-3p与c-kit mRNA表达水平、膀胱组织磷酸化（p）-ERK/ERK与c-kit表达水平均降低（P均<0.05）。与模型组、电针+miRNA-199a-3p抑制剂组相比，电针组大鼠膀胱组织病理损伤减轻，膀胱最大容量、膀胱基础压、漏尿点压力及血清COX-2、IL-18、iNOS水平均降低（P均<0.05），肌条自发性收缩频率、膀胱组织miRNA-199a-3p与c-kit mRNA表达水平、膀胱组织p-ERK/ERK与c-kit表达水平均升高（P均< 0.05）；miRNA-199a-3p抑制剂组大鼠膀胱组织病理损伤均加重，膀胱最大容量、膀胱基础压、漏尿点压力及血清COX-2、IL-18、iNOS水平均升高（P均<0.05），肌条自发性收缩频率、膀胱组织miRNA-199a-3p与c-kit mRNA表达水平及膀胱组织p-ERK/ERK与c-kit表达水平均降低（P均<0.05）。与模型组相比，miRNA-199a-3p抑制剂阴性对照组大鼠各指标水平差异无统计学意义（P均>0.05）。结论 电针关元穴可通过上调miRNA-199a-3p表达激活c-kit/ERK通路，减轻神经源性尿潴留大鼠膀胱组织炎症损伤，增强逼尿肌兴奋性，修复膀胱功能，改善尿潴留症状。 |
| 关键词: 微RNA-199a-3p c-kit/细胞外信号调节激酶通路 电针 关元穴 神经源性尿潴留 |
| DOI：10.16781/j.CN31-2187/R.20211145 |
| 投稿时间：2021-11-11修订日期：2022-03-07 |
| 基金项目:天津市教委科研计划项目（2018KJ028）. |
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| Therapeutic effect of electroacupuncture at Guanyuan acupoint on neurogenic urinary retention through microRNA-199a-3p regulating c-kit/ERK pathway in rats |
| LI Zhengfei,ZHANG Ren,ZHAO Guorui,KUANG Yao* |
(Department of Massage, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine/National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, Tianjin 300193, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the therapeutic effect of electroacupuncture at Guanyuan acupoint on neurogenic urinary retention through microRNA (miRNA)-199a-3p regulating receptor tyrosine kinase c-kit/extracellular signal-regulated kinase (ERK) pathway in rats. Methods Sixty model rats with neurogenic urinary retention were established by modified spinal cord transection method and divided into 5 groups by random number table method:model group, electroacupuncture group, miRNA-199a-3p antagomir (inhibitor) group, miRNA-199a-3p antagomir negative control group, and electroacupuncture+miRNA-199a-3p antagomir group, and another 12 SD rats were selected as sham group. After group intervention with drugs, the urodynamics of the rats were tested, and the maximum bladder volume, basal bladder pressure, and urine leakage point pressure were compared in each group; muscle strip experiment was used to detect the excitability of rat detrusor muscles and compare the spontaneous contraction frequency of muscle strips in each group; hematoxylin-eosin staining was used to detect the pathological and morphological changes of the bladder tissue of rats in each group; the kits were used to measure the serum levels of inflammatory cytokine cyclooxygenase 2 (COX-2), interleukin (IL-18) and inducible nitric oxide synthase (iNOS) of rats in each group; real-time fluorescence quantitative polymerase chain reaction experiment was used to detect the expression levels of miRNA-199a-3p and c-kit mRNA in the bladder tissue; Western blotting was used to detect the expression levels of c-kit/ERK pathway proteins in the bladder tissue. Results Compared with the sham group, the bladder tissue of the model group showed severe pathological damage, the maximum bladder volume, basal bladder pressure, urine leakage point pressure, serum COX-2, IL-18 and iNOS levels were significantly increased (all P<0.05), the spontaneous contraction frequency of muscle strips, the expression levels of miRNA-199a-3p and c-kit mRNA in bladder tissue, and the expression levels of phosphorylated (p)-ERK/ERK and c-kit in bladder tissue were significantly decreased (all P<0.05). Compared with the model group and the electroacupuncture+miRNA-199a-3p antagomir group, the pathological damage of the bladder tissue of rats in the electroacupuncture group was reduced, the maximum bladder volume, basal bladder pressure, urine leakage point pressure, serum COX-2, IL-18 and iNOS levels were decreased (all P<0.05), the spontaneous contraction frequency of muscle strips, the expression levels of miRNA-199a-3p and c-kit mRNA in bladder tissue, and the expression levels of p-ERK/ERK and c-kit in bladder tissue were increased (all P<0.05); the pathological damage in the miRNA-199a-3p antagomir group was aggravated, the maximum bladder volume, basal bladder pressure, urine leakage point pressure, serum COX-2, IL-18 and iNOS levels were increased (all P<0.05), the spontaneous contraction frequency of muscle strips, the expression levels of miRNA-199a-3p and c-kit mRNA, and the expression levels of p-ERK/ERK and c-kit were decreased (all P<0.05). Compared with the model group, there was no significant difference in the indicators in the miRNA-199a-3p antagomir negative control group (all P>0.05). Conclusion Electroacupuncture at Guanyuan acupoint can activate c-kit/ERK pathway by up-regulating the expression of miRNA-199a-3p, thereby reduce the inflammatory damage of bladder tissue in neurogenic urinary retention rats, enhance the excitability of detrusor, repair the bladder function, and improve the symptoms of urinary retention in rats. |
| Key words: microRNA-199a-3p c-kit/extracellular signal-regulated kinase pathway electroacupuncture Guanyuan acupoint neurogenic urinary retention |
