间充质干细胞对干燥综合征患者活化CD4＋T细胞miRNA表达谱的影响  

The effect of human umbilical cord mesenchymal stem cells on miRNA profile of activated peripheral blood CD4＋ T ceils from patient with primary Sjogren＇s syndrome

作　　者：龚邦东[1] 路臻豪[1] 景波[2] 黄家树[1] 阮光锋[1] 汤建平[1] 

机构地区：[1]同济大学附属同济医院风湿免疫科,上海200065 [2]同济大学附属同济医院干细胞临床转化中心,上海200065

出　　处：《中华风湿病学杂志》2015年第7期435-439,I0001,共6页Chinese Journal of Rheumatology

基　　金：基金项目：国家自然科学基金（81273295）;教育部留学回国人员基金（20101174）;上海市卫生局基金（2011276）

摘　　要：目的体外探讨人脐带来源间充质干细胞（Mscs）对DSS患者外周血活化CD4＋T细胞miRNA表达谱的影响。方法将pss患者外周血分选的CD4＋T细胞分为健康未活化组、pss未活化组、pSS活化组、MSC处理组和IFN．1．MSC处理组。对4组进行细胞计数；miRNA芯片检测CD4＋T细胞miRNA表达谱，定量PCR验证miRNA125b和miRNA155。组间均数比较用单因素方差分析，多重比较采用LSD法。结果MSC和IFN-γ-MSC均能以MSC依赖方式抑制活化CD4＋T细胞增殖，但两者之间比较差异无统计学意义。miRNA芯片发现pSS未活化组（对照为健康未活化组），pSS活化组（对照为psS未活化组），MSC处理组（对照为pSS活化组）和IFN．吖．MSC处理组（对照为MSC处理组）发生2倍以上变化的miRNAs分别为42、55、27、32个。进一步qPCR验证健康未活化组和上述4组的miRNA125b相对表达量分别为1．02±0．13、0．80±0．11、0．44±0．17、0．76±0．17和0．81±0．15（F=18．32，P〈0．01），miRNA155分别为1．5±0．8、3．9±1．3、8．4±2．6、10．1±4．2和11．2±5．0（F=26．65，P〈0．01）。结论MSCs能调节pSS患者活化CD4＋T细胞miRNA表达谱变化，能部分逆转活化CD4＋T细胞明显下调的miR．125b，后者可能在MSCs抑制活化CD4＋T细胞中起调控作用。Objective To investigate how human umbilical cord mesenchymal stem cells （MSCs） in vitro regulate the miRNA profile of activated peripheral blood CD4＋ T cells from patient with primary Sjogren＇s syndrome （pSS）. Methods Peripheral blood CD4＋ T cells from patient with pSS were sorted and divided into healthy naive group, pSS naive group, pSS activated group, MSC treatment group and MSC （pre-stimulated by IFN-γ） treatment group. CD4＋ T cells were counted. MiRNA microarray technology was used to detect the expression profile of CD4＋ T cells, and the expression of miRNA125b and miRNA155 was verified by real time quantification-polymerase chain reaction （RT-PCR）. Mean in groups were compared using ANOVA, and multiple comparisons were used with LSD method. Results Both MSCs and IFN-γ-MSCs could inhibit the proliferation of activated CD4＋ T cells in a MSC-dependent manner, but there was no significant difference between two groups. Microarray analysis found that the differentially enriched miRNAs in pSS naive （vs healthy naive）, pSS activation （vs pSS naive）, MSC treatment （vs pSS activation） and pre-IFN-＇,/MSC treatment （vs pSS activation） were 42 miRNAs, 56 miRNAs, 21 miRNAs and 24 miRNAs, respectively. Furthermore, the expressions of miRNA125b and miRNA155 were verified by RT-PCR and found that miRNA125b relative level in 5 groups was 1.02±0.13, 0.80±0.11, 0.44±0.17, 0.76±0.17 and 0.81±0.15 （F=18.32, P〈0.01）, and miRNAI$5 was 1.5±0.8, 3.9±1.3, 8.4±2.6, 10.1±4.2 and 11.2±5.0 （F=26.65, P〈0.01）. Conclusion MSCs can regulate miRNA profile of activated CD4~ T cells in peripheral blood of patient with pSS, and partiallyreverse down-regulated miR-125b in activated CD4~ T cells, which may play a regulatory role in inhibiting the activation of CD4~ T cells by MSCs.

关 键 词：干燥综合征 间质干细胞 CD4阳性T细胞 

分 类 号：R593.2[医药卫生—内科学]