纳米细菌诱导肾结石形成中肠道草酸降解菌变化初步研究  

Preliminary study on changes of oxalate-degrading bacteria in nanobacteria-induced kidney stone formation

作　　者：罗培跃 陈涛[1,2] 邹俊 李威 陈琪 钱彪 LUO Peiyue;CHEN Tao;ZOU Jun;LI Wei;CHEN Qi;QIAN Biao(Department of Urology,the First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,China)

机构地区：[1]赣南医学院第一附属医院泌尿外科,江西赣州341000 [2]赣州市泌尿男科重点实验室,江西赣州341000

出　　处：《陕西医学杂志》2023年第8期977-981,共5页Shaanxi Medical Journal

基　　金：江西省教育厅科学技术研究项目(GJJ2201403);赣南医学院资金资助项目(YJYB202111);赣南医学院研究生创新专项资金资助项目(YC2022-S959)。

摘　　要：目的:研究在纳米细菌诱导实验性大鼠肾结石形成过程中,大鼠肠道草酸降解菌的改变及其与肾结石形成的关系。方法:30只4周龄Wistar雄性大鼠适应性喂养1周后,按照随机数表法分为三组,对照组(NC组):1.2 ml 0.9%氯化钠溶液尾静脉注射+2 ml 0.9%氯化钠溶液灌胃;干扰组(NBT组):1.2 ml纳米细菌悬液尾静脉注射+2 ml四环素溶液灌胃;纳米细菌组(NB组):1.2 ml纳米细菌悬液尾静脉注射+2 ml 0.9%氯化钠溶液灌胃。标准饲养8周后处死,取双侧肾脏组织行HE、Pizzolatto’s染色,称取1 g回盲部新鲜粪便并提取DNA,使用qPCR法检测产甲酸草酸杆菌(Oxc)、乳杆菌属细菌(Lac)、双歧杆菌属细菌(Bif)和肠杆菌科细菌(Ent)四种主要肠道草酸降解细菌的基因表达量,留取腹主动脉血液及肾脏组织,流式细胞仪检测淋巴细胞分化情况,WB检测骨桥蛋白(OPN)及Tamm-Horsfall蛋白(THP)两种炎性蛋白在肾脏组织中的表达情况。结果:NC组肾小管管腔内未见明显晶体及炎症细胞浸润,NBT组部分肾小管扩张,肾小管管腔内可见不规则晶体及炎症细胞浸润,NB组肾小管明显扩张,肾小管管腔内可见大量不规则晶体,部分连接成片,可见大量炎症细胞浸润;三组大鼠粪便中均检测到四种肠道草酸降解菌,且各肠道草酸降解菌在三组大鼠盲肠中的占比相似,均为Oxc最高,约占半数,Lac、Ent次之,Bif最少,仅占约1%,三组大鼠粪便中各肠道草酸降解菌占比无统计学差异(均P>0.05),但三组粪便中肠道草酸降解菌的总量有统计学差异,NC组最高、NB组次之、NBT组最低,NC组与其余两组间的差异均有统计学意义(均P<0.01);流式细胞筛选结果提示三组大鼠血液淋巴细胞出现不同程度分化,Pearson相关性分析结果提示OPN与THP联系紧密,增长趋势具有一致性并且THP蛋白与CD4^(+)、CD8^(+)T淋巴细胞也具有强相关性。结论:纳米细菌诱导肾结石形成过程中肠道草酸降解菌总�Objective:To study the alteration of intestinal oxalate-degrading bacteria and its relationship with renal stone formation in nanobacteria-induced experimental rat kidney stone formation.Methods:Thirty 4-week-old Wistar male rats were acclimatized and fed for 1 week,then divided into three groups according to the random number table:control group(NC group,1.2 ml 0.9%NaCl solution tail vein injection+2 ml 0.9%NaCl solution gavage);interference group(NBT group,1.2 ml nanobacterial suspension tail vein injection+2 ml tetracycline solution gavage);nanobacterial group(NB group,1.2 ml nanobacterial suspension tail vein injection+2 ml 0.9%NaCl solution gavage).After standard feeding for 8 weeks and then executed,bilateral kidney tissues were taken for HE and Pizzolatto’s staining,1 g of fresh feces was weighed and DNA was extracted from the ileocecal region,and qPCR was used to detect the gene expression of four intestinal oxalate-degrading bacteria:Oxalobacter formigenes(Oxc),Lactobacillus(Lac),Bifidobacterium(Bif)and Enterobacteriaceae(Ent).Blood from the abdominal aorta and kidney tissues were retained and flow cytometry was used to detect lymphocyte differentiation,Western bot was used to detect the expression of two inflammatory proteins,osteopontin(OPN)and Tamm-Horsfall protein(THP)in the kidney tissues.Results:In the NC group,no obvious crystals and inflammatory cell infiltration were seen in the tubular lumen;in the NBT group,some tubules were dilated,and irregular crystals and inflammatory cell infiltration were seen in the tubular lumen;in the NB group,the tubules were significantly dilated,and a large number of irregular crystals were seen in the tubular lumen,some of which were connected in sheets and a large number of inflammatory cell infiltration was seen.Four intestinal oxalate-degrading bacteria were detected in the feces of all three groups of rats,and the proportion of each oxalate-degrading bacteria in the cecum of the three groups was similar,with Oxc being the highest,accounting for about half of

关 键 词：肾结石 纳米细菌 草酸代谢 肠道草酸降解菌 骨桥蛋白 TAMM-HORSFALL蛋白 

分 类 号：R692.4[医药卫生—泌尿科学]