补中愈淋汤对复发性尿路感染模型小鼠膀胱黏膜屏障及膀胱上皮细胞载菌量的影响  

Effects of Buzhong Yulin Decoction(补中愈淋汤)for Mice with Recurrent Urinary Tract Infectionon on Bladder Mucosal Barrier and Bacterial Load of Bladder Epithelial Cells

作　　者：尹浩 薛毅 张彪 金卓卉 朱娇丽 蒋毅 王晓芳 冯晨 金云云 金庆江 金庆雷 王鑫 YIN Hao;XUE Yi;ZHANG Biao;JIN Zhuohui;ZHU Jiaoli;JIANG Yi;WANG Xiaofang;FENG Chen;JIN Yunyun;JIN Qingjiang;JIN Qinglei;WANG Xin(Suzhou Hospital of Integrated Traditional Chinese and Western Medicine,Suzhou,215101;Yuexi Health Hospital of Wuzhong District)

机构地区：[1]苏州市中西医结合医院,江苏苏州215101 [2]苏州市吴中区越溪卫生院

出　　处：《中医杂志》2024年第22期2338-2346,共9页Journal of Traditional Chinese Medicine

基　　金：江苏省中医药科技发展计划项目(YB2020059);苏州市姑苏卫生人才计划人才科研项目(GSWS2022114);苏州市科技计划项目(SKY2023105,SKJY2021019);苏州市中西医结合科研基金(SKYD2023243);苏州市中西医结合医院院级课题立项项目(YJ2019012)。

摘　　要：目的探讨补中愈淋汤防治复发性尿路感染的可能作用机制。方法采用尿路致病性大肠杆菌(UPEC)UTI89菌株膀胱灌注法建立复发性尿路感染小鼠模型,造模成功小鼠随机分为模型组、抗生素组及补中愈淋汤低、高剂量组,每组6只。另取5只未造模的C57BL/6小鼠作为空白组。补中愈淋汤低、高剂量组分别予浓度为1.3、5.2 g/ml补中愈淋汤0.1 ml/10 g灌胃,抗生素组予5 mg/ml盐酸左氧氟沙星溶液0.1 ml/10 g灌胃,空白组及模型组予蒸馏水0.1 ml/10 g灌胃。各组均每天灌胃1次,连续7天。采用尿斑实验观察小鼠尿斑总数、中心尿斑数及尿斑总尿量;HE染色观察小鼠膀胱组织病理学变化;ELISA法检测血清炎症因子白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)含量;扫描电镜观察膀胱上皮细胞形态;免疫荧光法检测膀胱组织抗尿路斑块蛋白3A(UroPlakin 3A)蛋白水平及UPEC载菌量;涂板法检测尿菌量及膀胱上皮细胞内细菌量;RT-PCR法检测膀胱组织Ras相关蛋白11A(RAB11A)、Ras相关蛋白27B(RAB27B)mRNA水平;免疫印迹法检测膀胱组织微管相关轻链蛋白3(LC3)、P62蛋白水平。结果与空白组比较,模型组小鼠膀胱上皮细胞脱落,膀胱上皮细胞形态异常;膀胱上皮平均表面积减小,膀胱组织UroPlakin 3A蛋白及RAB11A、RAB27B mRNA水平降低;小鼠尿斑总数、中心尿斑数,膀胱组织UPEC载菌量,尿菌量,膀胱上皮细胞内细菌量,血清IL-1β、IL-6、TNF-α水平,膀胱组织LC3、P62蛋白水平均升高(P<0.05或P<0.01)。与模型组比较,补中愈淋汤低、高剂量组小鼠膀胱上皮细胞层完整、上皮细胞形态规则;膀胱上皮细胞平均表面积增大,膀胱组织UroPlakin 3A蛋白及RAB11A、RAB27B mRNA水平升高;小鼠尿斑总数、中心尿斑数,膀胱组织UPEC载菌量,尿菌量,膀胱上皮细胞内细菌量,血清IL-1β、IL-6、TNF-α水平,膀胱组织P62蛋白均降低(P<0.05或P<0.01),LC3蛋白表达�Objective To investigate the possible mechanism of Buzhong Yulin Decoction(补中愈淋汤)in the prevention and treatment of recurrent urinary tract infection.Methods The mouse models of recurrent urinary tract infection were established by uropathogenic Escherichia coli(UPEC)strain UTI89 by bladder perfusion,and the successful mouse models were randomly divided into a model group,an antibiotic group,and a low-and high-dose Buzhong Yulin Decoction group,with six mice in each group.In addition,5 C57BL/6 mice without modelling were taken as blank group.The low-and high-dose Buzhong Yulin Decoction groups received 0.1 ml/10 g of decoction by gavage,with concentrations of 1.3 g/ml and 5.2 g/ml,respectively;the antibiotic group received 0.1 ml/10 g of levofloxacin hydrochloride solution with 5 mg/ml by gavage;the blank and model groups received 0.1 ml/10 g of distilled water by gavage.Each group was gavaged once a day for 7 consecutive days.The total number of urine marks,the number of central urine marks,and the total urine volume of the urine marks were observed by the urine marking test;HE staining was used to observe the histopathological changes in the bladder of mice;serum levels of the inflammatory factors interleukin 1β(IL-1β),interleukin 6(IL-6)and tumour necrosis factorα(TNF-α)were detected by ELISA;the morphology of the epithelial cells of bladder was observed by scanning electron microscopy;immunofluorescence assay to detect bladder tissue anti-UroPlakin 3A protein level and UPEC bacterial load;the spread plate method to detect urinary bacterial load and bacterial load of bladder epithelial cells;RT-PCR method to detect Ras-related protein Rab-11A(RAB11A)and Ras-related protein Rab-27B(RAB27B)mRNA level in bladder tissue;immunoblotting to detect microtubule-associated protein 1 light chain3(LC3)and P62 protein levels in bladder tissue.Results Compared with the blank group,the bladder epithelial cell layers were lost and showed abnormal morphology in mice of the model group;bladder tissue UroPlakin 3A

关 键 词：复发性尿路感染 尿路致病性大肠杆菌 补中愈淋汤 膀胱组织 黏膜屏障 载菌量 尿路斑块蛋白 

分 类 号：R285.5[医药卫生—中药学]