中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2015年
7期
435-439
,共5页
龚邦东%路臻豪%景波%黄家树%阮光锋%汤建平
龔邦東%路臻豪%景波%黃傢樹%阮光鋒%湯建平
공방동%로진호%경파%황가수%원광봉%탕건평
干燥综合征%间质干细胞%CD4阳性T细胞
榦燥綜閤徵%間質榦細胞%CD4暘性T細胞
간조종합정%간질간세포%CD4양성T세포
Sj?gren's syndrome%Mesenchymal stem cells%CD4-positive T-lymphocytes
目的：体外探讨人脐带来源间充质干细胞（MSCs）对pSS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-γ-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-γ-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。
目的：體外探討人臍帶來源間充質榦細胞（MSCs）對pSS患者外週血活化CD4+T細胞miRNA錶達譜的影響。方法將pSS患者外週血分選的CD4+T細胞分為健康未活化組、pSS未活化組、pSS活化組、MSC處理組和IFN-γ-MSC處理組。對4組進行細胞計數；miRNA芯片檢測CD4+T細胞miRNA錶達譜，定量PCR驗證miRNA125b和miRNA155。組間均數比較用單因素方差分析，多重比較採用LSD法。結果 MSC和IFN-γ-MSC均能以MSC依賴方式抑製活化CD4+T細胞增殖，但兩者之間比較差異無統計學意義。 miRNA芯片髮現pSS未活化組（對照為健康未活化組）， pSS活化組（對照為pSS未活化組）， MSC處理組（對照為pSS活化組）和IFN-γ-MSC處理組（對照為 MSC處理組）髮生2倍以上變化的miRNAs分彆為42、55、27、32箇。進一步qPCR驗證健康未活化組和上述4組的miRNA125b相對錶達量分彆為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分彆為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）。結論 MSCs能調節pSS患者活化CD4+T細胞miRNA錶達譜變化，能部分逆轉活化CD4+T細胞明顯下調的miR-125b，後者可能在MSCs抑製活化CD4+T細胞中起調控作用。
목적：체외탐토인제대래원간충질간세포（MSCs）대pSS환자외주혈활화CD4+T세포miRNA표체보적영향。방법장pSS환자외주혈분선적CD4+T세포분위건강미활화조、pSS미활화조、pSS활화조、MSC처리조화IFN-γ-MSC처리조。대4조진행세포계수；miRNA심편검측CD4+T세포miRNA표체보，정량PCR험증miRNA125b화miRNA155。조간균수비교용단인소방차분석，다중비교채용LSD법。결과 MSC화IFN-γ-MSC균능이MSC의뢰방식억제활화CD4+T세포증식，단량자지간비교차이무통계학의의。 miRNA심편발현pSS미활화조（대조위건강미활화조）， pSS활화조（대조위pSS미활화조）， MSC처리조（대조위pSS활화조）화IFN-γ-MSC처리조（대조위 MSC처리조）발생2배이상변화적miRNAs분별위42、55、27、32개。진일보qPCR험증건강미활화조화상술4조적miRNA125b상대표체량분별위1.02±0.13、0.80±0.11、0.44±0.17、0.76±0.17화0.81±0.15（F=18.32，P<0.01），miRNA155분별위1.5±0.8、3.9±1.3、8.4±2.6、10.1±4.2화11.2±5.0（F=26.65，P<0.01）。결론 MSCs능조절pSS환자활화CD4+T세포miRNA표체보변화，능부분역전활화CD4+T세포명현하조적miR-125b，후자가능재MSCs억제활화CD4+T세포중기조공작용。
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 Sj?gren'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 na?ve (vs healthy na?ve), pSS activation (vs pSS na?ve), MSC treatment (vs pSS activation) and pre-IFN-γ 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 miRNA155 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 partially reverse 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.
