摘要
目的观察电针对脊髓损伤后神经源性膀胱(NB)大鼠TGF-β1/Smads信号通路及上皮-间质转化(EMT)相关蛋白表达的影响,探讨其改善NB纤维化的可能机制。方法36只雌性SD大鼠随机选取10只为假手术组,其余26只进行T8椎体下脊髓完全横断手术建立NB大鼠模型,将成模大鼠随机分为模型组和电针组,每组10只。电针组于术后19日起取“次髎”“中极”“三阴交”予电针干预,20 min/次,1次/d,连续7 d。观察大鼠一般状况,超声检测膀胱指标,尿流动力学检测膀胱功能,记录大鼠体质量、膀胱湿重并计算膀胱指数,HE染色观察膀胱组织形态,Masson染色观察膀胱组织纤维化程度及膀胱壁厚度,免疫组化检测膀胱组织E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)阳性表达,Western blot检测膀胱组织转化生长因子(TGF)-β1、p-Smad2/3、E-cadherin、N-cadherin、Vimentin蛋白表达。结果与假手术组比较,模型组大鼠膀胱上下径、左右径、前后径及膀胱容积明显增加(P<0.001),膀胱最大压力、漏尿点压力差、灌注时间、膀胱最大容量明显增加(P<0.001),膀胱指数明显升高(P<0.001),膀胱上皮细胞增厚且排列不规则,膀胱胶原容积分数、膀胱壁厚度明显增加(P<0.001),膀胱组织TGF-β1、p-Smad2/3、N-cadherin、Vimentin蛋白表达明显升高(P<0.01),E-cadherin蛋白表达明显降低(P<0.001)。与模型组比较,电针组大鼠膀胱上下径、左右径、前后径及膀胱容积明显减少(P<0.05),膀胱最大压力、漏尿点压力差、灌注时间、膀胱最大容量明显减少(P<0.05),膀胱指数明显降低(P<0.05),膀胱上皮细胞层厚度变薄、排列较整齐,膀胱胶原容积分数、膀胱壁厚度明显减少(P<0.05),膀胱组织TGF-β1、p-Smad2/3、N-cadherin、Vimentin蛋白表达明显降低(P<0.05),E-cadherin蛋白表达明显升高(P<0.05)。结论电针可能通过抑制TGF-β1/Smads信号通路活化减少膀胱上皮细胞EMT,降低膀胱组织纤维化程度,进而改善脊髓损伤后NB大鼠膀胱功能。
Objective To observe the effects of electroacupuncture(EA)on TGF-β1/Smads signaling pathway and epithelial-mesenchymal transition(EMT)related proteins in rats with neurogenic bladder(NB)after spinal cord injury;To explore the possible mechanism of EA in improving NB fibrosis. Methods Totally 36 female SD rats were randomly selected, with 10 rats in the sham-operation group and the remaining 26 rats undergoing complete transection of the spinal cord beneath the T8 vertebrae to establish a NB rat model. The modeling rats were randomly divided into model group and EA group, with 10 rats in each group. EA group was applied to “Ciliao”, “Zhongji” and “Sanyinjiao”, 20 min per time, once a day for 7 days. The general condition of the rats in each group was observed, the ultrasound index of the bladder was detected by ultrasound technique, the bladder function of the rats was detected by urodynamics, the body mass of the rats and the wet weight of the bladder were recorded, and the bladder index was calculated. HE staining was used to observe bladder tissue morphology, the degree of bladder tissue fibrosis and bladder wall thickness were detected by Masson staining. The positive expressions of E-cadherin, N-cadherin and Vimentin in bladder tissue were detected by immunohistochemistry. The protein expressions of TGF-β1, p-Smad2/3, E-cadherin, N-cadherin and Vimentin were detected by Western blot. Results Compared with the sham-operation group, the upper and lower diameter, anteroposterior diameter, transverse diameter, bladder volume of the model group significantly increased (P<0.001), the maximum bladder pressure, leak point pressure difference, perfusion time and maximum bladder capacity significantly increased (P<0.001), the bladder index increased significantly (P< 0.001), the bladder epithelial cells were thickened and arranged irregularly, the bladder collagen volume fraction and bladder wall thickness significantly increased (P<0.001), the expressions of TGF β1, p-Smad2/3, N-cadherin and Vimentin in bladder tissue increased (P<0.01), and the expression of E-cadherin decreased (P<0.001). Compared with the model group, the upper and lower diameter, anteroposterior diameter, transverse diameter, bladder volume of the EA group decreased significantly (P<0.05), the maximum bladder pressure, leak point pressure difference, perfusion time and maximum bladder capacity significantly decreased (P<0.05), the bladder index significantly decreased (P< 0.05), the thickness of the bladder epithelial cell layer became thinner and arranged more neatly, and the bladder collagen volume fraction and bladder wall thickness of the bladder were significantly reduced (P<0.05), the expressions of TGF-β1, p-Smad2/3, N-cadherin and Vimentin in bladder tissue significantly decreased (P<0.05), and the expression of E-cadherin significantly increased (P<0.05). Conclusion EA may reduce the EMT of bladder epithelial cells and decrease the degree of bladder tissue fibrosis by inhibiting the activation of the TGF-β1/Smads signaling pathway, thereby improving bladder function in NB rats after spinal cord injury.
作者
罗小精
田楚宁
展立芬
李芊
梁柔筠
肖露波
卓越
艾坤
许明
张泓
LUO Xiaojing;TIAN Chuning;ZHAN Lifen;LI Qian;LIANG Roujun;XIAO Lubo;ZHUO Yue;AI Kun;XU Ming;ZHANG Hong(College of Acupuncture&Moxibustion and Tui-na Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国中医药信息杂志》
2025年第10期120-126,共7页
Chinese Journal of Information on Traditional Chinese Medicine
基金
国家自然科学基金面上项目(82274666)
国家自然科学基金青年科学基金(82205255)
湖南中医药大学研究生创新项目(2024CX167、2024CX034)。
