期刊文献+

TGF-β1对大鼠高危角膜移植免疫排斥反应中植片IL-10表达的影响 被引量:1

Effects of TGF-β1 on expression of IL-10 in immunological rejection after high-risk penetrating keratoplasty
下载PDF
导出
摘要 目的研究重组腺相关病毒介导的转化生长因子-β1(recombinant adeno-associat-ed virus mediated transforming growth factor-β1,rAAV-TGF-β1)对大鼠高危角膜移植免疫排斥反应的抑制作用以及对白细胞介素-10(interleukin-10,IL-10)表达的影响。方法构建rAAV-TGF-β1真核表达质粒载体。40只角膜新生血管化的SD大鼠行穿透性角膜移植术。实验组(20只):角膜植片预浸泡在rAAV-TGF-β1(10×1012v.g.L-1)的达尔伯克必需基本培养液F12混合培养液中37℃孵育30min;对照组(20只):角膜植片预浸泡在0.1mol.L-1磷酸盐缓冲液中37℃孵育30min。裂隙灯观察TGF-β1对大鼠角膜植片存活时间、排斥反应指数的影响。术后第1周、第2周、第4周、第2个月对角膜植片行组织病理学检查,免疫组织化学方法检测角膜组织中IL-10的表达。结果对照组角膜植片平均存活时间为(10.40±2.45)d,实验组为(18.70±1.51)d,比对照组显著延长,差异具有显著统计学意义(P<0.01);术后第2周实验组角膜植片排斥反应指数(3.48±0.86)明显低于对照组(7.65±0.42),差异具有显著统计学意义(P<0.01)。IL-10在术后主要表达于角膜的上皮层与内皮层。术后各时期实验组IL-10的表达明显高于对照组,差异具有统计学意义(P<0.05)。结论rAAV-TGF-β1能显著延长角膜植片存活时间,抑制大鼠高危角膜移植免疫排斥反应,其机制与提高角膜组织中IL-10的表达有关。重组腺相关病毒是眼科基因治疗的有效载体。 Objective To study the immunosuppressive action of recombinant adeno-associated virus mediated transforming growth factor-β1 ( rAAV-TGF-β1 ) on the rat models of high-risk penetrating keratoplasty rejection and the effect of rAAV-TGF-β1 on the expression of interleukin-10 (IL-10). Methods The eukaryotic plasmid vector containing rAAV-TGF-β1 was constructed. Forty SD rats were operated with penetrating keratoplasty, and randomly divided into 2 groups, experimental group and control group ,20 animals in each group. Corneal neovascularization(CNV) were induced by alkaline cauterization of the central cornea. In experimental group, the donor cornea graft was incubated in 10 ×10^12 v. g · L^-1 rAAV-TGF-β1 dulbecco' s minimum essential medium mixed culture medium for 30 minutes at 37℃ ,and in control group,the donor graft was treated in phosphate;buffered solution for 30 minutes at 37℃ before surgery. Mean survival time and rejection index were clinically observed and compared with slit light microscope. The allografts were detected by histopathology, and expression of IL-10 was evaluated by immunhistochemistry at the 1st week,2nd week,4th week and 2nd month postoperatively. Resultu The mean survival time of control group were ( 10.40 ±2.45 ) d, were ( 18.70 ±1.51 ) d in experimental group, which was statistically significant compared with that of control group(P 〈 0.01 ). Rejection index (RI) of rAAV-TGF-β1 group was markedly lower compared with that of control group(P 〈0.01 ). The expression of IL-10 were increased at corneal epithelium and endothelium in experimental group compared with those in control group (P 〈 0. 05 ). Conclusions Treatment with rAAV-TGF-β1 can significantly prolong the survival time of corneal allograft and suppress the immunological rejection after high-risk penetrating keratoplasty, which may be associated with upregulating the expression of IL-10 in corneas. Recombinant adeno-associated virus is the effective vector for ophthalmologi
出处 《眼科新进展》 CAS 北大核心 2009年第1期12-17,共6页 Recent Advances in Ophthalmology
基金 国家自然科学基金资助(编号:30672290)~~
关键词 重组腺相关病毒 高危角膜移植 免疫排斥反应 转化生长因子-β1 白细胞介素-10 recombinant adeno-associated virus high-risk keratoplasty immuno- logical rejection transforming growth factor- β1 interleukin-10
  • 相关文献

参考文献10

  • 1Jonas JB, Rank RM, Budde WM. Immunologic graft reactions after allogenic penetrating keratoplasty [ J ]. Am J Ophthalmol 2002 ; 133 (4) :437-443. 被引量:1
  • 2Committee to revise the guide for the care and use of laboratory animals. Guide for the care and use of laboratory animals [ S ]. Washington D. C :National Academy Press; 1996:59-70. 被引量:1
  • 3赵敏,陈家祺,杨培增.鼠角膜碱烧伤的免疫学研究[J].中华眼科杂志,2000,36(1):40-42. 被引量:39
  • 4Larkin DF, Calder VL,Lightman SL. Indentification and characterization of cells infiltrating the graft and aqueous humour in rat allograft rejection[J]. Clin Exp Immunol 1997;107(2) :381- 391. 被引量:1
  • 5周炼红,胡燕华,汪道文.重组腺相关病毒经不同途径转染鼠眼球组织[J].眼科新进展,2004,24(3):182-184. 被引量:14
  • 6Borras T. Recent developments in ocular gene therapy[ J]. Exp Eye Res 2003 ; 75 ( 5 ) : 543 -652. 被引量:1
  • 7周炼红,邢怡桥,叶美洪,汪道文.重组腺相关病毒介导的报告基因在角膜内皮细胞中的表达[J].眼科研究,2006,24(2):169-171. 被引量:5
  • 8Maluccio M, Sharma V, Lagman M, Vyas S, Yang H, Li B, et al. Taerolimus enhances transforming growth factor-betal expression and promotes tumor progression [ J ]. Transplantation 2003 ;76 ( 3 ) :597-602. 被引量:1
  • 9Buer B J, Lanoue A, Franzke A, Gercia C, von Boehmer H, Serukhan A. Interleukin-10 secretion and imparied effector function of major histocompatibility complex class Ⅱ-restricted T cells anergized in vivo [ J ]. J Exp Med 1998 ; 187 ( 4 ) : 177-183. 被引量:1
  • 10Hara M,Kingsley CI,Niimi M,Gasperini S,Pauwels RA. IL-10 is required for regulatory T cells to mediate tolerance to alloantigens in vivo [ J]. J Immunol 2001 ; 166 (6) : 3789-3795. 被引量:1

二级参考文献22

  • 1Kessler PD, Podsakoff GM, Chen X, McQuiston SA, Colosi PC, Matelis LA,et al. Gene delivery to skeletal muscle results in sustained expression and systemic delivery of therapeutic protein[J]. Proc Natl Acad Sci USA 1996;93:14082-14087. 被引量:1
  • 2Hauswirth WW, Beaufrere L. Ocular gene therapy: quo vadis [J]? Invest Opthalmol Vis Sci 2000; 41:2821-2826. 被引量:1
  • 3Auricchio A, Hildinger M, O'Connor E, Gao GP, Wilson JM. Isolation of highly infections and pure adeno-associated virus Type 2 Vectors with a single-stop gravity-Flow column[J]. Hum Gene Ther 2001;12:71-76. 被引量:1
  • 4Tsai ML, Chen SL, Chou PI, Wen LY, Tsai JF, Tsao YP. Inducible adeno-associated virus vector-delivered transgene expression in corneal endothelium[J]. Invest Ophthalmol Vis Sci 2002;43:751-757. 被引量:1
  • 5Joyce NC, Meklir B,Joyce SJ, Zieske JD. Cell cycle protein expression and proliferative status in human corneal cells[J]. Invest Ophthalmol Vis Sci 1996;37:654-655. 被引量:1
  • 6Tanelian DL, Barry MA, Johnston SA, Le T, Smith GM. Controlled gene gun delivery and expression of DNA within the cornea[J]. Biotechniques 1997;23:484-488. 被引量:1
  • 7Shewring L, Collins L, Lightman SL, Hart S, Gustafsson K, Fabre JW. A nonviral vector system for efficient gene transfer to corneal endothelial cells via mebrane integrins[J]. Transplantation 1997;64:763-769. 被引量:1
  • 8Larkin DF, Oral HB, Ring CJ, Lemoine NR, George AJ. Adenvirus-mediated gene delivery to the corneal endothelium[J]. Transplantation 1996;37:1283-1293. 被引量:1
  • 9Lai CC, Wu WC, Xiao X, Tsai TC, Huan SJ, Chen TL, et al. Suppression of choroidal neovascularizaion by adeno-associated virus vector expressing angiostatin vector[J]. Invest Ophthalmol Vis Sci 2001;42:2401-2407. 被引量:1
  • 10Samulski RJ, Zhu X, Xiao X, Brook JD, Housman DE, Epstein N, et al. Targeted integration of adeno-associated virus (AAV)into human chromosome 19[J]. EMBOJ 1991;10:3940-3950. 被引量:1

共引文献51

同被引文献17

引证文献1

二级引证文献2

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部