[1]罗培跃,陈 涛,邹 俊,等.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[J].陕西医学杂志,2023,52(8):977-981.[doi:DOI:10.3969/j.issn.1000-7377.2023.08.008]
　LUO Peiyue,CHEN Tao,ZOU Jun,et al.Preliminary study on changes of oxalate-degrading bacteria in nanobacteria-induced kidney stone formation[J].,2023,52(8):977-981.[doi:DOI:10.3969/j.issn.1000-7377.2023.08.008]
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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
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
52
期数:
2023年8期
页码:
977-981
栏目:
基础研究
出版日期:
2023-08-05

文章信息/Info

Title:
Preliminary study on changes of oxalate-degrading bacteria in nanobacteria-induced kidney stone formation
作者:
罗培跃1; 2 ; 陈 涛1; 2; 邹 俊1; 2; 李 威1; 2 ; 陈 琪1; 2 ; 钱 彪1; 2
(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)
Author(s):
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)
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
Keywords:
Kidney stones;  Nanobacteria;  Oxalate metabolism;  Intestinal oxalate-degrading bacteria;  Osteopontin;  Tamm-Horsfall protein
分类号:
R 692.4
DOI:
DOI:10.3969/j.issn.1000-7377.2023.08.008
文献标志码:
A
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量降低,但各肠道草酸降解菌所占比例无明显改变。
Abstract:
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 the total,Lac and Ent being the second,and Bif being the least,accounting for only about 1% of the total,and there was no statistical difference in the proportion of each intestinal oxalic acid-degrading bacteria in the feces of the three groups(P>0.05).However,the total amount of intestinal oxalate-degrading bacteria in the feces of the three groups was statistically different,with the highest in the NC group,the second highest in the NB group,and the lowest in the NBT group,the differences between the NC group and the remaining two groups were statistically significant(P<0.01).The results of flow cytometric screening suggested that the blood lymphocytes in the three groups showed different degrees of differentiation,and the results of Pearson correlation analysis suggested that OPN was closely associated with THP and the growth trend was consistent and the THP protein was also strongly correlated with CD4+ and CD8+ T lymphocytes.Conclusion:The total amount of intestinal oxalate-degrading bacteria decreased during the formation of nanobacteria-induced kidney stones,but the proportion of each intestinal oxalic acid-degrading bacteria did not change significantly.

参考文献/References:

[1] Ansari H,Akhavan SA,Akhavan SM.Different approaches to detect nanobacteria in patients with kidney stones:An infectious cause or a subset of life[J].Urology Journal,2017,14(5):5001-5007.
[2] 钱 彪.纳米细菌诱导实验性大鼠肾结石形成机制的初步研究[D].武汉:华中科技大学,2019.
[3] 杨 恒,钱 彪,郑丽英,等.纳米细菌在肾结石形成过程中的作用研究[J].中国医药科学,2022,12(7):21-24.
[4] Wu W,Yang B,Ou L,et al.Urinary stone analysis on 12 846 patients:A report from a single center in China[J].Urolithiasis,2014,42(1):39-43.
[5] Ticinesi A,Nouvenne A,Chiussi G,et al.Calcium oxalate nephrolithiasis and gut microbiota:Not just a gut-kidney axis anutritional perspective[J].Nutrients,2020,12(2):241-349.
[6] Turroni S,Bendazzoli C,Dipalo SCF,et al.Oxalate-degrading activity in bifidobacterium animalis subsp lactis:Impact of acidic conditions on the transcriptional levels of the oxalyl coenzyme A(CoA)decarboxylase and formyl-CoA transferase genes[J].Applied and Environmental Microbiology,2010,76(16):5609-5620.
[7] Ferraz RRN,Marques NC,Froeder L,et al.Effects of Lactobacillus casei and Bifidobacterium breve on urinary oxalate excretion in nephrolithiasis patients[J].Urological Research,2009,37(2):95-100.
[8] Kaufman DW,Kelly JP,Curhan GC,et al.Oxalobacter formigenes may reduce the risk of calcium oxalate kidney stones[J].JASN,2008,19(6):1197-1203.
[9] Barr-Beare E,Saxena V,Hilt EE,et al.The interaction between enterobacteriaceae and calcium oxalate deposits[J].PloS One,2015,10(10):139575.
[10] 黄美泰,伍颖君,郑永波,等.严重腹腔感染时血、粪纳米细菌的检测及其与SIRS关系的实验研究[J].中华普通外科学文献:电子版,2013,7(4):284-288.
[11] Qian B,Zheng L,Wang Q,et al.Correlation between ApoE gene polymorphisms and the occurrence of urolithiasis[J].Experimental and Therapeutic Medicine,2015,9(1):183-186.
[12] Corniere N,Thomson RB,Thauvin S,et al.Dominant negative mutation in oxalate transporter SLC26A6 associated with enteric hyperoxaluria and nephrolithiasis[J].Journal of Medical Genetics,2022,59(11):1035-1043.
[13] Khan SR,Canales BK,Dominguez-Gutierrez PR.Randall plaque and calcium oxalate stone formation:role for immunity and inflammation[J].Nature Reviews Nephrology,2021,17(6):417-433.
[14] 钟方红,皮特俊章,钟 琴,等.中药对高尿酸血症患者肠道菌群影响的研究进展[J].陕西中医,2021,42(2):265-268.
[15] Amin R,Asplin J,Jung D,et al.Reduced active transcellular intestinal oxalate secretion contributes to the pathogenesis of obesity-associated hyperoxaluria[J].Kidney International,2018,93(5):1098-1107.
[16] Miller AW,Choy D,Penniston KL,et al.Inhibition of urinary stone disease by a multi-species bacterial network ensures healthy oxalate homeostasis[J].Kidney International,2019,96(1):180-188.
[17] Zhang Y,Zhang Q.Relationship between tetracycline antibiotic susceptibility and genotype in oral cavity Lactobacilli clinical isolates[J].Antimicrobial Resistance and Infection Control,2019,8:27.
[18] Mukherjee R,Dutta D,Patra M,et al.Nanonized tetracycline cures deadly diarrheal disease shigellosis in mice,caused by multidrug-resistant shigella flexneri 2a bacterial infection[J].Nanotechnology Biology and Medicine,2019,18:402-413.
[19] Zhang Y,Tan P,Zhao Y,et al.Enterotoxigenic escherichia coli:Intestinal pathogenesis mechanisms and colonization resistance by gut microbiota[J].Gut Microbes,2022,14(1):2055943.
[20] Bunker JJ,Erickson SA,Flynn TM,et al.Natural polyreactive IgA antibodies coat the intestinal microbiota[J].Science,2017,358:6361.
[21] Brown EM,Kenny DJ,Xavier RJ.Gut microbiota regulation of T cells during inflammation and autoimmunity[J].Annual Review of Immunology,2019,37:599-624.
[22] 刘应莉,王艳荣,张秋瓒.肠道菌群-过氧化物酶体增殖物激活受体信号通路及其在代谢相关脂肪性肝病发病中的作用研究进展[J].陕西医学杂志,2021,50(9):1166-1169.
[23] 李 江,卢金芳.防石合剂对草酸钙肾结石患者ESWL术后尿OPN水平的影响[J].陕西中医,2017,38(2):209-210.
[24] 易建华,李 雯.脂多糖诱导的急性炎症小鼠模型血清促炎因子与抑炎因子表达研究[J].陕西医学杂志,2023,52(4):395-398.

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备注/Memo

备注/Memo:
基金项目: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)
更新日期/Last Update: 2023-08-07