间充质干细胞对小鼠活化B细胞微小核糖核酸155和肿瘤坏死因子-α表达的影响  

The effects of mesenchymal stem cells on miRNA-155 and tumor hecrosis factor-α expression in activated B cells from mice

作　　者：龚邦东[1] 路臻豪[1] 景波[2] 黄家树[1] 阮光峰[1] 刘劼[2] 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]同济大学干细胞中心

出　　处：《国际免疫学杂志》2015年第3期199-202,共4页International Journal of Immunology

基　　金：国家自然科学基金资助（81273295）

摘　　要：目的 研究间充质干细胞（MSCs）对小鼠活化B细胞微小核糖核酸155（miRNA-155）和肿瘤坏死因子-α（TNF-α）表达的影响,探讨MSCs对B细胞的调控机制.方法 以小鼠脾脏原代B细胞为研究模型,在B细胞受体（BCR）刺激下,与人脐带来源的MSCs共培养3d后,收集悬浮的B细胞,进行细胞计数,实时定量PCR检测B细胞miRNA-155和TNF-α mRNA水平,酶联免疫吸附试验（ELISA）法检测共培养上清中细胞因子免疫球蛋白G（IgG）、TNF-α、转化生长因子β（TGF-β）、前列腺素E2（PGE2）、白细胞介素（IL）-10和吲哚胺2,3-双加氧酶（IDO）水平.结果 B细胞活化后增殖明显,MSCs能轻微抑制B细胞增殖（F =5.39,P＜0.05）.B细胞活化后miRNA-155和TNF-α mRNA均上调明显,MSCs能部分逆转miRNA-155和TNF-α mRNA的升高（F=18.85,P＜0.01和F=10.26,P＜0.05）.B细胞活化后共培养上清TNF-α、IgG升高明显,MSCs能部分缓解TNF-α和IgG的升高（F=27.94,P＜0.01和F=4.81,P＜0.05）.在MSCs和B细胞共培养上清中,PGE2升高尤为明显,其次是IDO.结论 MSC可能通过可溶性因子PGE2和IDO抑制活化B细胞miRNA-155和TNF-α的表达,MSCs可能通过抑制B细胞治疗免疫疾病发挥重要作用.Objective To study the effects of mesenchymal stem cells （MSCs） on the microRNA-155 （miRNA-155） and tumor necrosis factor-α（TNF-α） of activated B cells,and explore the regulation mechanisms of MSCs on B cells.Methods We used primary B cells from mouse spleen as the research models.B cells were co-cultured with human umbilical cord derived MSCs for three days in the B-cell receptor （BCR） stimulation.The suspension cells were collected and counted.Real-time PCR was used to determine the miRNA-155 and TNF-α mRNA levels in B cells.Enzyme-linked immunosorbent assay （ELISA） was used to inspect supernatant immunoglobulin G （IgG） and cytokines including TNF-α,transforming growth factor （TGF-β）,prostaglandin E2 （PGE2）,interleukin-10 （IL-10）,and indoleamine 2,3-dioxygenase （IDO）.Results B cells showed significant proliferation after activation,which can be slightly inhibited by MSCs （F =5.39,P 〈0.05）.MiRNA-155 and TNF-o mRNA in B cells were significantly upregulated after B-cell activation,which can be partially reversed by MSCs （F =18.85,P 〈 0.01 and F =10.26,P 〈 0.05）.TNF-α and IgG in co-culture supernatant were significantly increased after B cell activation,and MSCs can partially alleviate the elevated TNF-α and IgG （F =27.94,P 〈 0.01 and F =4.81,P 〈 0.05）.In co-culture supernatants,PGE2 increasesd markedly,followed by IDO.Conclusion MSC can downregulate miRNA-155 and TNF-α expression in activated B cells possibly via PGE2 or IDO.This might play some important role in MSC treatment for immune disorders.

关 键 词：间充质干细胞 B细胞 微小核糖核酸155 肿瘤坏死因子-Α 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]