体外条件下人骨髓间充质干细胞与NZBWF1鼠T淋巴细胞的免疫调节  

Immunoregulation effects of bone marrow mesenchymal stem cells on splenic T-lymphocytes in NZBW F1 mice in vitro

作　　者：宋晓丽[1] 李晶[1] 裘影影[1] 殷玉俊[1] 尤海燕[1] 芮金兵[1] 汤郁[1] 

机构地区：[1]江苏大学附属医院风湿科,江苏省镇江市212001

出　　处：《中国组织工程研究与临床康复》2009年第36期7128-7132,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：江苏省科技厅社会发展项目(BS2005043);镇江市科技局社会发展项目(SH2004040)~~

摘　　要：背景:近年研究发现系统性红斑狼疮患者的间充质干细胞存在异常,自体造血干细胞移植治疗系统性红斑狼疮复发率高,有学者提出系统性红斑狼疮不仅是造血干细胞病,也是间质干细胞病。目的:探讨人骨髓间充质干细胞在体外对NZBWF1鼠脾脏T淋巴细胞的免疫调节作用。设计、时间及地点:细胞共培养体外实验,于2008-03/09在江苏大学附属医院中心实验室完成。材料:健康人骨髓由江苏大学附属医院研究生自愿捐献。SPF级20周龄NZBWF1雌鼠9只,购自南京大学动物模式研究所。方法:Percoll密度梯度离心法分离培养人骨髓间充质干细胞,取P3代细胞用于实验。20周龄NZBWF1小鼠适应性饲养至24周龄时,无菌分离脾淋巴细胞,在刀豆球蛋白A刺激下,将T淋巴细胞与骨髓间充质干细胞分别以不同比例(1:0.01,1:0.02,1:0.1)共培养72h,并设立对照组。主要观察指标:MTT法检测T淋巴细胞增殖,流式细胞仪检测T淋巴细胞凋亡和CD69,CD28表达,实时荧光定量PCR检测T淋巴细胞白细胞介素10、干扰素γmRNA水平。结果:与对照组比较,在体外人骨髓间充质干细胞能抑制NZBWF1鼠T淋巴细胞的增殖,且呈剂量依赖性(P<0.05)。与对照组比较,T淋巴细胞与骨髓间充质干细胞比例为1:0.02和1:0.1时,骨髓间充质干细胞可抑制T淋巴细胞的凋亡,并促进T淋巴细胞干扰素γmRNA表达,抑制白细胞介素10mRNA表达;T淋巴细胞与骨髓间充质干细胞比例为1:0.01时,T淋巴细胞凋亡情况和干扰素γ、白细胞介素10mRNA表达无明显差异(P>0.05)。结论:人骨髓间充质干细胞体外在一定比例下可能通过抑制NZBWF1鼠T淋巴细胞的增殖和凋亡、干预Th1/Th2细胞因子mRNA表达,继而下调T淋巴细胞的免疫功能。BACKGROUND： Present studies have shown that mesenchymal stem cells （MSCs） of systemic lupus erythematosus patients are abnormal. Autologous hematopoietic stem cell transplantation for treating systemic lupus erythematosus has high recurrence rate. Some scholars have proposed that systemic lupus erythematosus is not only a hematopoietic stem cell disease, but also a MSCs disease. OBJECTIVE： To investigate the immunoregulation effects of human bone marrow mesenchymal stem cells （BMSCs） on splenic T-lymphocytes of NZBW F1 mice in vitro. DESIGN, TIME AND SETTING： The co-culture in vitro study was conducted at the Central Laboratory of Affiliated Hospital of Jiangsu University from March to September 2008. MATERIALS： Bone marrow was obtained from healthy postgraduates from the Affiliated Hospital of Jiangsu University. A total of 9 SPF grade female NZBW F1 mice aged 20 weeks were purchased from the Animal Mode Institute of Nanjing University. METHODS： Human BMSCs were separated by the Percoll density gradient centrifugation. At passage 3, BMSCs were used in this study. Splenic lymphocytes were isolated from 24-week NZBW F1 female mice following breeding. Under Concanavalin A stimulation, T-lymphocytes were treated with BMSCs according to different proportions （1：0.01, 1：0.02, 1：0.1） for 72 hours. A control group was set. MAIN OUTCOME MEASURES： The proliferation of T-lymphocytes was assessed by MTT assay. Flow cytometry was used to analyze the T-lymphocyte apoptosis and surface markers CD69, CD28. The expression of interleukin-10 and interferon-y mRNA of T-lymphocytes were detected by real-time quantitative PCR. RESULTS： Compared with the control group, human BMSCs could inhibit the proliferation of T-lymphocytes in NZBW F1 mice in a dose-dependent fashion （P 〈 0.05）. Compared with the control group, when the proportion of T-lymphocytes and BMSCs was 1：0.02 and 1：0.1, BMSC could inhibit the apoptosis of T-lymphocytes, and promote the mRNA expression of interferon-y mRNA

关 键 词：间充质干细胞 红斑狼疮 系统性 T淋巴细胞 免疫调节 

分 类 号：R394.2[医药卫生—医学遗传学]