摘要
目的探讨脊神经双侧离断神经源膀胱(NB)幼鼠模型的膀胱形态结构功能变化及转化生长因子β1(TGF-β1)通路相关蛋白的表达。方法选取SD雌性幼鼠64只,其中32只行L6+S1神经双侧离断建立NB模型,32只行假手术作为对照。神经离断和假手术3、6、12、24周后行膀胱测压检查。收集对应膀胱组织行马松染色和天狼猩红染色分析胶原纤维变化,免疫组织化学染色分析TGF-β1、Smad2、Smad6蛋白变化,采用Western blot检测TGF-β1受体Ⅰ的蛋白水平变化。结果各NB神经离断组膀胱不稳定,存在膀胱漏尿点压(BLPP),膀胱无明显排尿收缩压;神经离断3、6、12周和24周鼠的膀胱基础压(22.10±2.51)、(18.20±1.52)、(31.20±2.82)、(41.10±3.41)cmH2O(1 cmH2O=0.098 kPa)和膀胱容量(22.30±1.72)、(49.10±5.54)、(30.30±2.68)、(13.50±1.52)mL均明显高于假手术组[(3.51±0.45)cmH2O和(0.52±0.04)mL],差异均有统计学意义(均P<0.01)。神经离断组膀胱组织横切面大小为先增大(3、6周)后变小挛缩(12、24周),膀胱壁厚度先变薄(3、6周)后增厚(12、24周);膀胱质量随离断时间持续增加。马松染色结果显示,神经离断大鼠的膀胱壁中正常纤维结缔组织逐渐紊乱,膀胱壁分层结构逐渐解体,平滑肌肥大增厚,肌间胶原纤维逐渐增多,染色变深。天狼猩红检测示神经离断24周的膀胱组织中Ⅲ型胶原纤维明显增多,Ⅲ型/Ⅰ型胶原纤维比例(3.14±0.71)明显高于对应的假手术组(0.88±0.21),差异有统计学意义(t=7.48,P<0.01)。免疫组织化学染色发现,神经离断膀胱组织中TGF-β1和Smad2表达随着离断时间逐渐升高,通路抑制性蛋白Smad6随离断时间逐渐降低。Western blot显示TGF-β1受体Ⅰ蛋白水平在12、24周的表达分别高于对应的假手术组1.3、1.6倍(t=6.06、14.45,均P<0.01)。结论脊神经双侧离断导致的幼鼠NB,收缩功能瘫痪,膀胱内压逐渐增高,膀胱正常结构逐渐解体,膀胱纤维化通路TGF-β1/
Objective To investigate changes in the bladder morphological structure and function and the expression of transforming growth factor-beta1(TGF-β1)pathway-related proteins in the bilateral spinal nerve amputated neurogenic bladder(NB)rat.Methods A total of 64 female SD rats were included,and 32 of them underwent bilateral spinal nerve L6+S1 amputation to construct the NB model and the others were used as sham operation controls.Rats in both NB and control groups received bladder cystometry 3,6,12,24 weeks after corresponding operation.Collagen fibers in their bladder tissues were detected by Masson staining and Sirius scarlet staining.TGF-β1,Smad2 and Smad6 proteins were checked by immunohistochemical staining.TGF-β1 receptorⅠprotein was measured by Western blot.Results Bladders in the NB group were instable,with bladder leak point pressure(BLPP)and underactive voiding pressures.The basal pressure[(22.10±2.51),(18.20±1.52),(31.20±2.82),(41.10±3.41)cmH2O(1 cmH2O=0.098 kPa)]and bladder volume[(22.30±1.72),(49.10±5.54),(30.30±2.68),(13.50±1.52)mL]of the NB rats at 3,6,12 and 24 weeks were significantly higher than those of the sham operation controls[(3.51±0.45)cmH2O and(0.52±0.04)mL],and the difference were significant(all P<0.05).The bladder size and thickness in the NB group firstly increased(3,6 weeks)and then decreased(12,24 weeks),but the bladder weight increased continuously.Masson staining showed disordered fibrous connective tissues,disintegrated layered bla-dder wall,hypertrophied smooth muscle tissues and deposited intramuscular collagen on the nerve-amputated bladder wall.Sirius scarlet staining suggested that 24 weeks after nerve amputation,collagenⅢincreased greatly,and the ratio of typeⅢ/Ⅰcollagen fibers(3.14±0.71)was significantly higher than that in the sham group(0.88±0.21)(t=7.48,P<0.01).According to the immunohistochemical staining results,the expressions of TGF-1βand Smad2 increased while the pathway inhibitory protein Smad6 decreased with time in the NB group.Western blo
作者
陈燕
马源
何育霖
刘二鹏
邢栋
杨兴欢
朱文
王庆伟
窦启锋
张会清
文建国
Chen Yan;Ma Yuan;He Yulin;Liu Erpeng;Xing Dong;Yang Xinghuan;Zhu Wen;Wang Qingwei;Dou Qifeng;Zhang Huiqing;Wen Jianguo(Institute of Clinic Medicine,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Pediatric Urodynamic Center and Department of Urology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Urology,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Pro-vince,China)
出处
《中华实用儿科临床杂志》
CSCD
北大核心
2020年第17期1336-1340,共5页
Chinese Journal of Applied Clinical Pediatrics
基金
国家自然科学基金(U1904208,81670689)。
