未成熟树突状细胞在大鼠高危角膜移植免疫排斥反应中的作用  

Effects of immature dendritic cell in rat high-risk corneal transplantation rejection

作　　者：付燕[1] 杜安杰[1] 高晓唯[2] 李文静[2] 李霞[2] 赵旭东[2] 

机构地区：[1]石河子大学医学院,新疆维吾尔自治区石河子市830002 [2]解放军474医院全军眼科中心,新疆维吾尔自治区乌鲁木齐市830013

出　　处：《眼科新进展》2013年第2期130-134,共5页Recent Advances in Ophthalmology

基　　金：军区医药卫生科研项目资助(编号:CLZ11JA25)~~

摘　　要：目的研究供体骨髓来源的未成熟树突状细胞(immature dendritic cell,imDC)对大鼠高危角膜移植免疫排斥反应的抑制作用,探讨imDC抑制排斥反应的机理。方法以60只SD大鼠为受体,30只Wistar大鼠为供体,碱烧伤建立同种异体高危角膜移植模型。受体SD大鼠随机分为对照组、imDC组、成熟树突状细胞(mature dendritic cell,mDC)组,每组20只;对照组:术前7d经尾静脉注射PBS液0.1mL;imDC组:术前7d经尾静脉注射体外培养的供体骨髓来源的imDC1×106个;mDC组:术前7d经尾静脉注射体外培养的供体骨髓来源的mDC1×106个。术后每天裂隙灯观察各组角膜植片存活情况并评分;于术后3d、7d、14d每组随机处死4只大鼠,取角膜植片行HE染色,Westernblot分析检测各组大鼠脾脏CD4+CD25+调节性T细胞表面特异性标志转录因子Foxp3编码的Scurfin蛋白的表达。结果 imDC组角膜植片存活时间为(19.6±1.1)d,较对照组的(9.5±0.9)d和mDC组的(7.0±1.6)d明显延长,差异均有统计学意义(均为P<0.01);Westernblot检测结果显示,术后3d、7d、14dimDC组Foxp3编码的Scurfin蛋白的表达(相对吸光度值分别为2.21±0.76、2.31±0.68、2.24±0.18)明显高于对照组(相对吸光度值分别为1.41±0.12、1.47±0.25、1.68±0.09)和mDC组(相对吸光度值分别为1.43±0.14、1.58±0.41、1.46±0.13),差异均有统计学意义(均为P<0.05)。结论应用体外培养的供体来源的imDC可以抑制大鼠同种异体高危角膜移植免疫排斥反应,而诱导CD4+CD25+调节性T细胞的产生是其降低大鼠角膜移植术后免疫排斥反应的机制之一。Objective To investigate the inhibitive effects of immature dendritic cells （ imDC） of donor bone marrow on rat high-risk corneal transplantation rejection,and explore the mechanism of imDC inhibitive effects.Methods Wistar and SD rats were respectively used as donors and acceptors.Sixty SD rats were randomly divided into control group,imDC group and mature dendritic cells （ mDC） group,20 cases in each group.Corneal neovascularization was induced by alkaline burn.In the control group,SD rats were only injected 0.1mL PBS via tail vein before operation; In the imDC group,donor bone marrow-derived imDC of 1 × 10 6 were injected into SD rats via tail vein before operation; In the mDC group,donor bone marrow-derived mDC of 1 × 10 6 were injected into SD rats via tail vein before operation.The survival conditions of corneal grafts were observed and scored by slit lamp at each day after operation.In each group,four rats were killed at 3 days,7 days and 14 days after transplantation,the corneal grafts were stained by HE,and Western blot was used to detect Scurfin protein expression of CD4 ＋ CD25 ＋ T cells.Results The survival time of allograft rejection in imDC group was （ 19.6±1.1） days,which was longer than that in control group （ 9.5±0.9） days and mDC group （ 7.0±1.6） days,there were significant differences（ all P 0.01） .The Scurfin protein expressions of CD4 ＋ CD25 ＋ T cells in imDC group （ relative optical density were 2.21±0.76, 2.31±0.68 and 2.24±0.18） were significantly higher than those in control group （ relative optical density were 1.41±0.12,1.47±0.25 and 1.68±0.09） and mDC group （ relative optical density were 1.43±0.14,1.58±0.41 and 1.46±0.13） ,there were statistical difference （ all P 0.05） .Conclusion The pretreatment with imDC can prolong the survival time of rat corneal graft after transplantation.The mechanism of immune tolerance induced by imDC can be inhibited by T lymphocytes responsiveness by regulatory T cells.

关 键 词：未成熟树突状细胞 角膜移植 免疫耐受 调节性T细胞 

分 类 号：R699.2[医药卫生—泌尿科学]