全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

嘌呤霉素肾病模型肾组织微小RNA的差异性表达及雷至胶囊的干预作用

Keywords: 微小RNA,雷至胶囊,嘌呤霉素氨基核苷肾病,足细胞,蛋白尿

Full-Text   Cite this paper   Add to My Lib

Abstract:

目的:观察嘌呤霉素氨基核苷(puromycinaminonucleoside,PAN)肾病模型鼠肾组织微小RNA(microRNAs,miRNAs)差异性表达的特征及其与足细胞裂隙膜(sliddiaphragm,SD)关键结构分子nephrin,podocin和骨架蛋白synaptopodin表达的关系;阐明雷至胶囊(Leizhicapsule,LZC)在体内通过调控模型鼠肾组织miRNAs差异性表达而改善足细胞nephrin,podocin,synaptopodin表达,减少蛋白尿的机制。方法:将50只雄性Wistar大鼠随机分为空白组(A)、模型组(B)、雷至胶囊组(C,5mL·kg-1·d-1)、雷公藤多苷组(D,10mL·kg-1·d-1)和缬沙坦(E,7.5mL·kg-1·d-1)组。除A组外,其余各组大鼠一次性经颈静脉注射PAN(100mg·kg-1)而建立PAN肾病模型。造模后第2天灌胃给药,A,B组大鼠用生理盐水干预,共10d。造模前及造模后第3,9天称量各组大鼠体重,并检测24h尿蛋白排泄量(urinaryproteinration,Upro)。造模后第11天,处死全部大鼠,采集血液和肾组织,观察血清生化指标血清白蛋白(albumin,Alb),血清肌酐(serumcreatinine,Scr),血清尿素氮(bloodureanitrogen,BUN),肾小球超微结构(足细胞足突形态)以及肾组织dicer酶,nephrin,podocin,synaptopodin表达;同时,借助生物芯片(biochip)技术,分析肾皮质miRNAs差异性表达的特征,并且,借助荧光实时定量聚合酶链反应(Real-timePCR)验证rno-miR-23a,rno-miR-300-3p,rno-miR-24,rno-miR-30c等的差异性表达量。结果:在PAN诱导下,模型鼠出现蛋白尿、肾功能减退、低白蛋白血症、足细胞足突融合;在模型鼠肾组织中,dicer酶影响足细胞nephrin,podocin,synaptopodin表达;上调rno-miR-23a,rno-miR-300-3p表达,下调rno-miR-24,rno-miR-30c表达;差异性表达的miRNAs包括rno-miR-24,rno-miR-30c,rno-miR-23a,rno-miR-300-3p等。雷至胶囊能改善PAN肾病模型鼠的一般情况、蛋白尿、血清BUN、足细胞足突融合;还能减少模型鼠肾组织dicer酶表达,增加足细胞nephrin,podocin和synaptopodin表达;减弱在模型鼠肾组织中上调的rno-miR-23a,rno-miR-300-3p,增强在模型鼠肾组织中下调的rno-miR-24,rno-miR-30c等。结论:PAN在体内通过dicer酶/miRNAs差异性表达途径而影响模型鼠肾组织miRNAs表达,干预足细胞nephrin,podocin,synaptopodin表达,破坏足细胞结构和功能,产生蛋白尿;雷至胶囊可以减少PAN肾病模型鼠蛋白尿,其机制可能是干预dicer酶/miRNAs差异性表达途径,调控足细胞nephrin,podocin,synaptopodin表达,改善足细胞的结构与功能。

References

[1]  Poy M N,Eliasson L,Krutzfeldt J, et al. A pancreatic islet-specific microRNA regulates insulin secretion[J]. Nature,2004,432 (7014):226.
[2]  Van Rooij E,Sutherland L B,Qi X,et al. Control of stress-dependent cardiac growth and gene expression by a microRNA[J]. Science,2007,316 (5824):575.
[3]  Ho J,Ng K H,Rosen S,et al. Podocyte-specific loss of functional microRNAs leads to rapid glomerular and tubular injury[J]. J Am Soc Nephrol,2008,19 (11):2069.
[4]  吴汉利,孙伟,万毅刚,等.雷至胶囊对阿霉素肾病大鼠肾小球系膜基质增生抑制作用的实验研究[J].中国中西医结合肾病杂志,2010,11(1):18.
[5]  吴汉利,孙伟. 雷至胶囊对阿霉素肾病大鼠足细胞分子nephrin的影响[J]. 中国医院药学杂志,2010,30 (24):2076.
[6]  刘丽华,朱春芳,欧周罗.单次静脉注射嘌呤霉素氨基核苷所致大鼠肾病模型的优化及评价[J].复旦学报:医学版,2005,32(4):488.
[7]  Bartel D P. MicroRNAs: genomics,biogenesis,mechanism,and function[J]. Cell,2004,116 (2):281.
[8]  Croce C M,Calin G A. miRNAs,cancer,and stem cell division[J]. Cell,2005,122 (1):6.
[9]  Lewis B P,Burge C B,Bartel D P. Conserved seed pairing,often flanked by adenosines,indicates that thousands of human genes are microRNA targets[J]. Cell,2005,120 (1):15.
[10]  Willenbrock H,Salomon J,Sokilde R,et al. Quantitative miRNA expression analysis: comparing microarrays with next-generation sequencing[J]. RNA,2009,15 (11):2028.
[11]  Sun Y, Koo S,White N,et al. Development of a micro-array to detect human and mouse microRNAs and characterization of expression in human organs[J]. Nucleic Acids Res,2004,32 (22):e188.
[12]  Tian Z,Greene A S,Pietrusz J L,et al. MicroRNA-target pairs in the rat kidney identified by microRNA microarray,proteomic,and bioinformatic analysis[J]. Genome Res,2008,18 (3):404.
[13]  Shi S,Yu L,Chiu C,et al. Podocyte-selective deletion of dicer induces proteinuria and glomerulosclerosis[J]. J Am Soc Nephrol,2008,19 (11):2159.
[14]  Inokuchi S,Shirato I,Kobayashi N,et al. Re-evaluation of foot process effacement in acute puromycin aminonucleoside nephrosis[J]. Kidney Int,1996,50 (4):1278.
[15]  Kawachi H,Koike H,Kurihara H,et al. Cloning of rat nephrin: expression in developing glomeruli and in proteinuric states[J]. Kidney Int,2000,57 (5): 1949.
[16]  Nakatsue T,Koike H,Han G D,et al. Nephrin and podocin dissociate at the onset of proteinuria in experimental membranous nephropathy[J]. Kidney Int,2005,67 (6):2239.
[17]  Chhabra R,Dubey R,Saini N. Cooperative and individualistic functions of the microRNAs in the miR-23a~27a~24-2 cluster and its implication in human diseases[J]. Mol Cancer,2010,9:232.
[18]  Chhabra R,Adlakha Y K,Hariharan M,et al. Upregulation of miR-23a~27a~24-2 cluster induces caspase-dependent and-independent apoptosis in human embryonic kidney cells[J]. PLoS One,2009,4 (6):e5848.
[19]  Haraldsson B,Jeansson M. Glomerular filtration barrier[J]. Curr Opin Nephrol Hypertens,2009,18 (4):331.
[20]  Patrakka J,Tryggvason K. New insights into the role of podocytes in protein uria[J]. Nat Rev Nephrol,2009,5 (8):463.
[21]  Huber T B,Kottgen M,Schilling B,et al. Interaction with podocin facilitates nephrin signaling[J]. J Biol Chem,2001,276 (45):41543.
[22]  Saleem M A,Ni L,Witherden I,et al. Co-localization of nephrin,podocin and the actin cytoskeleton:evidence for a role in podocyte foot process formation[J]. Am J Pathol,2002,161 (4):1459.
[23]  Johnstone D B,Holzman L B. Clinical impact of research on the podocyte slit diaphragm[J]. Nat Clin Pract Nephrol,2006,2 (5):271.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133