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Clusterin抑制人肾近曲小管上皮HK-2细胞的凋亡 被引量:2

Clusterin Prevents Apoptosis in Human Proximal Tubular Cells HK-2
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摘要 Clusterin是一种硫酸糖蛋白.最近研究发现,clusterin具有抗凋亡作用,同时对肾细胞具有保护作用,但抗凋亡的具体机制仍不清楚.为研究clusterin及其不同功能区域在人肾近曲小管上皮HK-2细胞中的抗凋亡作用,构建了含有全长及缺失前导序列的clusterin重组质粒(分别命名为pIRES2-EGFP/cluac和pIRES2-EGFP/clubc).将重组质粒转染人肾近曲小管上皮HK-2细胞后,检测转染细胞中clusterin的表达及其抗Na2SeO3(10μmol/L)诱导的凋亡作用.Western印迹显示,转染pIRES2-EGFP/cluac的HK-2细胞培养上清及细胞裂解液中均可检测到clusterin蛋白表达,但转染pIRES2-EGFP/clubc的HK-2细胞仅在裂解液中检测到clusterin,在培养上清液中未检测到该蛋白表达.流式细胞术检验显示,HK-2/clubc细胞实验组出现明显凋亡峰,而HK-2/cluac细胞组则未见凋亡;两组的凋亡百分率之间也存在显著性差异(P<0.05).以Cy3标记的Annexin V染色后于荧光显微镜下观察细胞凋亡情况与FCM检测结果基本一致.上述结果证明,clusterin有明显的抑制人肾近曲小管上皮HK-2细胞凋亡的作用;clusterin前导序列是其发挥抗凋亡作用的必需功能区域,提示clusterin抗凋亡作用是通过细胞外途径产生的. Clusterin is a sulfated glycoprotein and prevents apoptosis in kidney cells.To evaluate the effect of clusterin containing different domains,the mechanism of preventing apoptosis was studied when the different domains of clusterin were over-expressed in human proximal tubular cells(PTC) HK-2.The plasmid pIRES2-EGFP was used to generate the clusterin expression constructs with full-length or without the leader sequence(designated as pIRES2-EGFP/cluac,pIRES2-EGFP/clubc,respectively).Western blotting was employed to compare clusterin protein expression levels in the lysate and supernatant of HK-2 cells which were transfected by clusterin.When HK-2 cells were induced to produce by Na2SeO3,the clusterin's protective role was examined by flow cytometry(FCM) and fluorescence microscope in HK-2 cells.Clusterin expression was detected in the lysate and supernatant of pIRES2-EGFP/cluac transfected HK-2 cells;while clusterin was only found in lysate of pIRES2-EGFP/clubc transfected HK-2 cells,whereas it was not found in supernatant.Its anti-apoptotic property in HK-2 cells was confirmed by FCM and fluorescence microscope.It was apparent that clusterin played an important role in preventing apoptosis in human proximal tubular cells,and the presence of the leader sequence was necessary for clusterin's anti-apoptotic function.
出处 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2008年第9期833-838,共6页 Chinese Journal of Biochemistry and Molecular Biology
关键词 CLUSTERIN 肾小管 HK-2细胞 凋亡 clusterin renal tubular HK-2 cells apoptosis
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参考文献15

  • 1Blaschuk O, Burdzy K, Fritz I B. Purification and characterization of a cell - aggregating factor ( clusterin ) , the major glycoprotein in ram rete testis fluid[J]. J Biol Chem, 1983,.258(12) :7714-7720
  • 2De Silva H V, Harmony J A, Stuart W D. Apolipoprotein J: Structure and tissue distribution [J]. Biochemistry, 1990, 29(22) : 5380-5389
  • 3Dvergsten J, Manivel J C, Correa-Rotter R, et al. Expression of clusterin in human renal diseases [J]. Kidney Int, 1994,45(3): 828-835
  • 4Rosenberg M E, Silkensen J. Clusterin: physiologic and pathophysiologic considerations [J]. Int J Biochem Cell Biol, 1995, 27(7) :633-645
  • 5Rosenberg M E, Silkensen J. Clusterin and the kidney [ J]. Exp Nephrol, 1995,3(1) :9-14
  • 6Parczyk K, Pilarsky C, Rachel U, et al. Gp80 ( clusterin; TRPM- 2 ) mRNA level is enhanced in human renal clear cell carcinomas [J]. J Cancer Res Clin Oncol, 1994,120(3) : 186-188
  • 7Rosenberg M E, Dvergsten J, Correa-Rotter R. Clusterin: an enigmatic protein recruited by diverse stimuli[J]. J Lab Clin Med,1993,121(2):205-214
  • 8Humphreys D T,Carver J A, Easterbrook-Smith S B, et al. Clusterin has chaperone-like activity similar to that of small heat shock proteins [J]. J Biol Chem, 1999,274( 11 ) :6875-6881
  • 9French L E, Polla L L, Tschopp J, et al. Membrane attack complex (MAC) deposits in skin are not always accompanied by S-protein and elusterin[J]. J Invest Dermatol, 1992,98(5): 758-763
  • 10Murphy B F, Davies D J, Morrow W, et al. Localization of terminal complement components S-protein and SP-40, 40 in renal biopsies [J]. Pathology, 1989,21(4):275-278

同被引文献37

  • 1王晓华,王笑云,熊明霞,张飞飞,王宁宁,李瑾,何东元,杨俊伟.大黄蒽醌提取物缓解小鼠肾组织纤维化作用的实验研究[J].中华肾脏病杂志,2006,22(6):360-363. 被引量:8
  • 2王青秀,吴纯启,杨红莲,荆淑芳,金城,肖小河,廖明阳.大黄中游离蒽醌对HK-2细胞系的毒性作用研究[J].中国新药杂志,2007,16(3):189-192. 被引量:15
  • 3Fritz IB, Burdzy K, Stchell B, et al. Ram rete testis fluid contains a protein (clusterin) which influences cell-cell interactions in vitro[J].Biol Reprod,1983,28(5) :1173.
  • 4Murphy BF,Kirszbaum I.,Walker ID,et aI. SP 40,40,a newly i dentil'led normal human serum protein found in the SCSb 9 corn plex of complement and in the immune deposits in glomerulone phritis[J]. J Clin Invest, 1988,81(6) : 1858.
  • 5Jene DE, Tschopp J. Molecular structure and funetional charac terization of a human complement eyto[ysis inhibitor found in blood and seminal plasma: identity to sulfated glycoprotein-2 a constituent of rat testis fluid[J]. Proe Natl Acad Sei USA, 1989, 86(18):7123.
  • 6Elias-Sonnenschein LS, Bertram I., Visser PJ. Relationship be- tween genetic risk factors and markers for Alzheimer's disease pa- thology [J]. Biomark Med. 2012,6(4):477.
  • 7B Shannan, M Seifert, K Leskov, et al. Challenge anti promise: roles for clusterin in pathogenesis, progression and therapy of cancer [J]. Cell Death and Differentiation,2006,13( 1): 12.
  • 8Rosenberg ME,Dvergsten J ,Correa-Rotter R. Clusterin: an enig- matic protein recruited by diverse stimuli [J]. J Lab Clin Med, 1993,121(2) :205.
  • 9Rosenberg ME, Girton R, Finkel D, et al. Apolipoprotein clusterin prevents a progressive glomerulopathy of aging [J].Mol Cell Bi- ol,2002,22(6) : 1893.
  • 10Rosenberg ME,Silkensen J. Clusterin and the kidney [J]. Exp Nephrol,1995,3(1) :9.

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