加味春泽汤调控NF-κB/TGF-β_(1)信号通路对脊髓损伤神经源性膀胱尿潴留大鼠膀胱纤维化及逼尿肌功能的影响  

Effect of Modified Chunzetang on Bladder Fibrosis and Detrusor Function in Rats with Neurogenic Bladder Urinary Retention Induced by Spinal Cord Injury via Regulating NF-κB/TGF-β_(1) Signaling Pathway

作　　者：徐振华 李彦杰 任亚峰[3] 刘昊源 朱博超 刘娟 XU Zhenhua;LI Yanjie;REN Yafeng;LIU Haoyuan;ZHU Bochao;LIU Juan(Henan Provincial Hospital of Traditional Chinese Medicine/Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450053,China;Rehabilitation Medicine College,Henan University of Chinese Medicine,Zhengzhou 450008,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南省中医院/河南中医药大学第二附属医院,郑州450053 [2]河南中医药大学康复医学院,郑州450008 [3]河南中医药大学第一附属医院,郑州450002

出　　处：《中国实验方剂学杂志》2025年第5期95-103,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省中医药科学研究重大专项(2023ZYZD07);2023年度河南省中医学“双一流”创建科学研究专项(HSRP-DFCTCM-2023-3-26);河南省中医药拔尖人才培养项目专项(2022ZYBJ15);中原英才计划中原青年拔尖人才项目(豫组通[2021]44号)。

摘　　要：目的:探讨加味春泽汤对脊髓损伤神经源性膀胱尿潴留大鼠膀胱纤维化及逼尿肌功能的治疗作用及其作用机制。方法:该研究以改良的Hassan Shaker脊髓横断法建立脊髓损伤神经源性膀胱尿潴留模型,选取脊髓损伤行为学评分0分的大鼠进入后续实验;经选取的大鼠随机分为模型组(生理盐水灌胃)、中药低剂量组(加味春泽汤14.4 g·kg^(-1)灌胃)、中药高剂量组(加味春泽汤28.8 g·kg^(-1)灌胃)、阳性药组[核转录因子-κB(NF-κB)抑制剂二硫代氨基甲酸吡咯烷(PDTC)0.05 g·kg^(-1)腹腔注射]、联合组(PDTC 0.05 g·kg^(-1)腹腔注射+加味春泽汤28.8 g·kg^(-1)灌胃),每日1次,干预4周。实验结果采用BL-420s生物机能采集系统统计尿流动力学指标,离体膀胱快速称质量并采用逼尿肌牵拉实验记录各组大鼠膀胱最小收缩张力及频率;苏木素-伊红(HE)染色及马松(Masson)染色对比各组大鼠逼尿肌病理形态及组织纤维化改变;免疫组化检测α-平滑肌肌动蛋白(α-SMA)表达水平;蛋白免疫印迹法(Western blot)检测各组大鼠膀胱组织中核转录因子-κB p65(NF-κB p65)、核转录因子-κB抑制蛋白α(IκBα)、转化生长因子-β(1 TGF-β_(1))、Ⅰ型胶原蛋白(ColⅠ)和Ⅲ型胶原蛋白(ColⅢ)蛋白表达;酶联免疫吸附测定法(ELISA)检测大鼠血清IL-6、IL-1β、TNF-α含量。结果:与假手术组比较,模型组大鼠漏尿点压力显著降低(P<0.01),膀胱质量、膀胱收缩张力、膀胱最大容量及膀胱顺应性均增加(P<0.05,P<0.01),HE染色可见膀胱上皮细胞排列紊乱,黏膜层及肌层中性粒细胞浸润等病理表现较为明显,固有层结构破坏,肌纤维排列紊乱,间质增宽、组织水肿较为明显;Masson染色可见模型组大鼠膀胱壁中胶原纤维沉积较多、逼尿肌纤维化程度较重,视野内逼尿肌含量有所减少;模型组大鼠膀胱组织NF-κB p65、TGF-β_(1)、IκBα、ColⅠ和ColⅢ蛋白表达显著升高(P<0.01),血�Objective:To investigate the therapeutic effect and mechanism of modified Chunzetang on bladder fibrosis and detrusor function in rats with neurogenic bladder urinary retention induced by spinal cord injury.Methods:In this study,an improved Hassan Shaker spinal cord transection method was used to establish a model of neurogenic bladder urinary retention induced by spinal cord injury,and rats with a spinal cord injury behavior score of 0 were selected for follow-up experiments.The selected rats were randomly divided into a model group(normal saline gavage),low-dose traditional Chinese medicine(TCM)group(gavage of 14.4 g·kg^(-1)modified Chunzetang),high-dose TCM group(gavage of 28.8 g·kg^(-1)modified Chunzetang),positive drug group[intraperitoneal injection of 0.05 g·kg^(-1)nuclear transcription factor-κB(NF-κB)inhibitor pyrrolidine dithiocarbamate(PDTC)],and combination group(intraperitoneal injection of 0.05 g·kg^(-1)PDTC+gavage of 28.8 g·kg^(-1)modified Chunzetang).The rats in these groups were administrated with corresponding drugs once a day for four weeks.The BL-420s biofunction acquisition system was used in the experiment to calculate the urodynamic indexes,and the isolated bladder was quickly weighed.The detrusor traction experiment was used to record the minimum bladder contraction tension and frequency in each group.The pathological morphology and tissue fibrosis of detrusor in each group observed by Hematoxycin-eosin(HE)staining and Masson staining were compared.The expression level ofα-smooth muscle actin(α-SMA)was detected by immunohistochemistry.Western blot was used to detect the protein expression of NF-κB p65,nuclear transcription factor-κB suppressor proteinα(IκBα),transforming growth factor-β_(1)(TGF-β_(1)),typeⅠcollagen(ColⅠ),and typeⅢcollagen(ColⅢ)in bladder tissue of rats in each group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the changes in serum levels of IL-6,IL-1β,and TNF-α.Results:Compared with that in the sham operation group,the pressure at th

关 键 词：神经源性膀胱 膀胱纤维化 核转录因子-κB(NF-κB) 加味春泽汤 转化生长因子-β_(1)(TGF-β_(1)) 

分 类 号：R242[医药卫生—中医临床基础] R285[医药卫生—中医学] R694