CAJ | 학술논문

背景：目前已有临床应用造血干细胞联合间充质干细胞输注治疗再生障碍性贫血的报道。目的：观察人脂肪间充质干细胞对再生障碍性贫血T淋巴细胞分泌功能的调节作用。<br>　　方法：从健康人脂肪中提取并培养人脂肪间充质干细胞，从再生障碍性贫血患者外周血中分离T淋巴细胞，将两者共培养后，以健康志愿者的T淋巴细胞单独培养及与人脂肪间充质干细胞共培养做对照。ELISA法检测上清白细胞介素2、白细胞介素4、白细胞介素10和γ-干扰素的水平，Realtime RCR和Western blot测定T-bet和GATA-3的表达。<br>　　结果与结论：共培养7 d后，间充质干细胞+再生障碍性贫血T淋巴细胞组Th1类细胞因子γ-干扰素、白细胞介素2水平明显低于再生障碍性贫血T淋巴细胞组(P<0.05)，Th2类细胞因子白细胞介素4、白细胞介素10水平明显高于再生障碍性贫血T淋巴细胞组(P<0.05)；间充质干细胞+再生障碍性贫血 T淋巴细胞组T-bet mRNA及蛋白表达水平均低于再生障碍性贫血T淋巴细胞组，GATA-3 mRNA及蛋白水平均高于再生障碍性贫血 T淋巴细胞组。提示人脂肪间充质干细胞在体外对再生障碍性贫血患者T淋巴细胞具有免疫调节作用，其机制可能是通过调节转录因子T-bet和GATA-3的表达从而抑制Th细胞向Th1细胞方向分化。
배경：목전이유림상응용조혈간세포연합간충질간세포수주치료재생장애성빈혈적보도。목적：관찰인지방간충질간세포대재생장애성빈혈T림파세포분비공능적조절작용。<br>　　방법：종건강인지방중제취병배양인지방간충질간세포，종재생장애성빈혈환자외주혈중분리T림파세포，장량자공배양후，이건강지원자적T림파세포단독배양급여인지방간충질간세포공배양주대조。ELISA법검측상청백세포개소2、백세포개소4、백세포개소10화γ-간우소적수평，Realtime RCR화Western blot측정T-bet화GATA-3적표체。<br>　　결과여결론：공배양7 d후，간충질간세포+재생장애성빈혈T림파세포조Th1류세포인자γ-간우소、백세포개소2수평명현저우재생장애성빈혈T림파세포조(P<0.05)，Th2류세포인자백세포개소4、백세포개소10수평명현고우재생장애성빈혈T림파세포조(P<0.05)；간충질간세포+재생장애성빈혈 T림파세포조T-bet mRNA급단백표체수평균저우재생장애성빈혈T림파세포조，GATA-3 mRNA급단백수평균고우재생장애성빈혈 T림파세포조。제시인지방간충질간세포재체외대재생장애성빈혈환자T림파세포구유면역조절작용，기궤제가능시통과조절전록인자T-bet화GATA-3적표체종이억제Th세포향Th1세포방향분화。
BACKGROUND:Clinical infusion of hematopoietic stem cells and mesenchymal stem cells for treatment of aplastic anemia has been reported. OBJECTIVE:To investigate the effect of human adipose-derived mesenchymal stems cells on the secretion function of T lymphocytes of aplastic anemia patients. METHODS:Human adipose-derived mesenchymal stems cells were extracted from healthy human adipose tissues. T-lymphocytes were harvested from peripheral blood of patients with aplastic anemia by density gradient centrifugation. Human adipose-derived mesenchymal stems cells were co-cultured with T-lymphocytes. The levels of interleukin-2, interleukin-4, interleukin-10 and interferon-γwere detected by enzyme linked immunosorbent assay. T-bet and GATA-3 levels were examined by real-time PCR and western blot. RESULTS AND CONCLUSION:The levels of Th1 type cytokines interferon-γand interleukin-2 in the co-culture group were significantly lower than those in the T-lymphocyte group (P<0.05). But the levels of Th2 type cytokines interleukin-4 and interleukin-10 in the co-culture group were significantly higher than those in the T-lymphocyte group (P<0.05). The T-bet mRNA and protein levels in the co-culture group were significantly lower than those in the T-lymphocyte group, while the GATA-3 mRNA and protein levels were significantly higher in the co-culture group. Human adipose-derived mesenchymal stems cells can mediate an immunoregulation effect on T-lymphocytes of aplastic anemia patients in vitro, which is possibly related with the inhibition of Th1-dominant response due to the disorder of T-bet and GATA-3 gene expression.