异体脂肪间充质干细胞对卵巢早衰患者外周血调节性T细胞的影响  

Effect of allogeneic adipose derived mesenchymal stem cells on regulatory T cells in peripheral blood of patients with premature ovarian failure

作　　者：宋开静 何援利[1] 蔡慧华 张冬梅[1] 黄睿淳 孙冬华[1] SONG Kai-jing;HE Yuan-li;CAI Hui-hua;ZHANG Dong-mei;HUANG Rui-chun;SUN Dong-hua(Department of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China;Department of Obstetrics and Gynecology, Guangdong General Hospital, Guangzhou 510080j China)

机构地区：[1]南方医科大学珠江医院妇产科,广州510282 [2]广东省人民医院妇产科,广州510080

出　　处：《解放军医学杂志》2018年第4期294-298,共5页Medical Journal of Chinese People's Liberation Army

基　　金：广东省医学科学技术研究基金项目(A2016034)~~

摘　　要：目的探讨异体脂肪间充质干细胞(h ADSCs)对卵巢早衰(POF)患者外周血调节性T细胞(Treg)的影响。方法采用0.1%Ⅰ型胶原酶消化法从脂肪组织中分离培养h ADSCs,并进行成脂成骨分化诱导以及表面分子鉴定。采用密度梯度离心法分离POF患者外周血单个核细胞(PBMCs)。在植物凝集素(PHA)的刺激下,将PBMCs分别与1×10~4、2×10~4、1×10~5个hADSCs共培养72h,CCK-8法检测淋巴细胞的增殖率,流式细胞技术测定CD4^+CD25^+Foxp3^+Treg的比例,实时荧光定量PCR检测Foxp3^+mRNA的表达水平。结果 h ADSCs高表达CD90、CD105,低表达CD34、CD45,具有成脂成骨分化能力。CCK-8检测结果显示,与对照组相比,各数量级的h ADSCs均能明显抑制淋巴细胞的增殖(P<0.001)。流式检测结果显示,各数量级的hADSCs均能促进CD4^+CD25^+Foxp3^+Treg的增殖(P<0.05)。qPCR检测结果显示,与对照组相比,各实验组均能显著上调Foxp3^+mRNA的表达(P<0.001)。结论 hADSCs能发挥免疫调节作用,促进卵巢早衰患者外周血Treg的增殖。Objective To investigate the effect of allogeneic adipose derived mesenchymal stem cells（h ADSCs） on regulatory T cells（Tregs） in peripheral blood of patients with premature ovarian failure（POF）. Methods h ADSCs were isolated from adipose tissue using 0.1% ty pe Ⅰ collagenase digestion. Adipogenic, osteogenic differentiation and surface molecular characterization were also performed. Peripheral blood mononuclear cells（PBMCs） of POF patients were isolated by density gradient centrifugation. PBMCs were co-cultured with 1×10^4, 2×10^4 and 1×10^5 hADSCs for 72 hours under the stimulation of phytohemagglutinin（PHA）. The proliferation rate of lymphocytes was measured by CCK-8 method, the proportion of CD4^＋ CD25^＋Foxp3^＋ Treg cells was measured by flow cytometry. Real-time fluorescent quantitative PCR was used to detect the mRNA level of Foxp3^＋. Results h ADSCs were positive for CD90, CD105 and negative for CD34, CD45, and has adipogenic and osteogenic differentiation ability. The results of CCK-8 showed that hADSCs could significantly inhibit the proliferation of lymphocytes compared with that in control group（P〈0.001）. Flow cytometry showed that hADSCs could promote the proliferation of CD4^＋CD25^＋ Foxp3^＋ Treg cells（P〈0.05）. The results of q PCR showed that Foxp3^＋ mRNA expression was obviously up-regulated in all experimental groups compared with that in control group（P〈0.001）. Conclusion h ADSCs can play an immune regulatory role and promote the proliferation of Tregs in peripheral blood of patients with premature ovarian failure.

关 键 词：脂肪间充质干细胞 卵巢早衰 调节性T细胞 FOXP3 

分 类 号：R711.59[医药卫生—妇产科学]