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细胞间粘附分子-1的表达与胰岛细胞移植关系的实验研究

Intracerebral transplantation of human erythropoietin gene-modified mesenchymal stem cells derived from human umbilical cord blood alleviates hypoxic ischemic brain injury in neonatal rats
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摘要 目的探讨经门静脉胰岛细胞移植(PIT)后,细胞间粘附分子-1(ICAM-1)的表达与移植免疫排斥反应的关系,为胰岛细胞移植术后排斥反应的临床治疗提供理论依据。方法30只Wistar糖尿病大鼠随机分为三组,A组:糖尿病模型组(n=10),不做任何处理;B组:生理盐水组(n=10),采用腹腔注射生理盐水2ml/d;C组:免疫抑制剂组(n=10),采用腹腔注射CsA(3mg·kg^-1.d^-1)+MMF(20mg·kg^-1.d^-1);全部采取SD大鼠胰岛细胞→Wistar大鼠糖尿病动物模型。观察:①各组胰岛细胞移植术后第1、4、7、10、30、60天大鼠外周血血糖的变化;②流式细胞仪(Elite Coulter Company US)技术检测胰岛细胞移植术后外周血粘附分子ICAM-1的表达情况;③免疫组化方法检测各组胰岛细胞移植术后第10、60天,肝脏中ICAM-1的表达水平。结果①血糖值变化:术前各组间血糖值比较无统计学差异(P=0.669);B、C组血糖在术后第7或10天达最低水平,之后逐渐上升;任何两组间比较有统计学差异。②外周血ICAM-1的表达情况:术前各组间ICAM-1值比较无统计学差异(P=0.754);术后C组在第7或10天达最低水平,以后逐渐上升,最后接近B组水平;任何两组间比较有统计学差异。③肝脏粘附分子表达:术后10d和术后60d比较:A组(P=0.816)无统计学差异,B组(P=0.855)无统计学差异,C组(P=0.000)有统计学差异;术后10d:任何两组间均存在统计学差异(P〈0.05);术后60d:B组与C组无统计学差异,A组同任何组均存在统计学差异(P〈0.05)。结论PIT后粘附分子的变化与免疫排斥反应的发生有很大的相关性,进而和移植物功能有一定的相关性,且粘附分子表达水平的升高可以预示免疫排斥反应的发生。 Objective To investigate the therapeutic effect of intracerebral transplantation of human erythropoietin (EPO) gene-modified mesenchymal stem cells (MSCs) derived from human umbilical cord blood (UCB) on hypoxic-ischemic brain damage (HIBD) in neonatal rats.Methods MSCs were isolated from human UCB by density gradient centrifugation,and identified by flow cytometry (FCM). pcDNA3-EPO was introduced into the human UCB-derived MSCs by lipofectamine and the expression of exogenous EPO was examined by RT-PCR and Western blot. Forty-two 7-day-old SD rats with HIBD were divided into 3 groups:control group (A group,n = 12),MSCs transplant group (B group,n = 12),and EPO gene-modified MSCs transplant group(C group,n = 18). On the 3rd day after HIBD,MSCs were injected into the left cortex of neonatal rats. Seven days after transplantation,6 rats of C group were sacrificed to investigated the survival and migration of the transplanted cells by immunohistochemtry. On the 1st,7th,14th,21st and 28th days after transplantation,nervous function of 3 groups were evaluated by modified neurological severity score (mNSS). Results MSCs were isolated from full-term newborn UCB. FACS analysis showed that MSCs were positive for CD29,CD44,and CD105 and negative for CD34,CD106,and HLA-DR. The results of RT-PCR and Western blot analyses indicated that the exogenous EPO could be successfully expressed in human UCB-derived MSCs. Immunocytochemistry analysis of brain tissue sections showed that the transplanted human UCB-derived MSCs could survive and migrate around from the center of transplant site. mNSS showed that the score of C group was significantly smaller than control group on the 14th,21st,and 28th days (P〈 0.05 or P〈 0.01),and mNSS of B group was significantly smaller than control group on the 21st and 28th days (P〈 0.05). Compared with B group,the score of C group was significantly smaller on the 21st and 28th day(P〈 0.05). Conclusion Transplantation of EPO gene-modified MSC
出处 《中华细胞与干细胞移植(电子版)》 2009年第2期18-22,共5页
关键词 胰岛细胞 移植 粘附分子 糖尿病大鼠 Erythropoietin Mesenchymal stem cells Human umbilical cord blood Hypoxic-ischemic brain damage Transplant
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