CAJ | 학술논문

目的：探讨间充质干细胞（ MSC）对原发免疫性血小板减少症（ ITP）小鼠血小板数目的影响及其机制。方法采用腹腔注射大鼠抗小鼠CD41抗体（ MWReg30单抗）的方法诱导BALB/c小鼠产生ITP后，取20只作为实验组，20只作为对照组。之后通过鼠尾静脉给实验组小鼠注射MSC 2×107个/只。注射后5 d、7 d、14 d，测定实验组与对照组小鼠外周血血小板数；14 d时应用逆转录聚合酶链反应（ RT-PCR）检测小鼠外周血单个核细胞（ PBMC）中转录因子T-bet和GATA-3 mRNA的表达；应用酶联免疫法测定Th1类细胞因子IFN-γ、IL-2及Th2类细胞因子IL-4、IL-10的水平。结果7 d、14 d后，注射MSC实验组小鼠血小板值[（588．0±81．6）×109/L、（623．0±78．9）×109/L]明显高于未注射MSC对照组[（317．0±90．1）×109/L、（288．0±87．8）×109/L]（P＜0．05）；14 d时，实验组PBMC中T-bet mRNA表达水平低于对照组[（0．04±0．03） vs （0．27±0．05）]（P＜0．05）；GATA-3 mRNA的表达水平高于对照组[（0．14±0．04） vs （0．07±0．05）]（P＜0．05）；实验组Th1类细胞因子IFN-γ、IL-2水平[（3．1±1．7） pg/ml、（3．2±2．1） pg/ml]明显低于对照组[（10．3±4．8） pg/ml、（16．3±5．7） pg/ml]（P＜0．05）；实验组外周血 Th2类细胞因子 IL-4、IL-10水平[（88．6±15．2） pg/ml、（38．3±11．8） pg/ml]明显高于对照组[（32．7±5．7） pg/ml、（22．1±3．4） pg/ml]（P＜0．05）。结论 MSC可以有效提升ITP小鼠的血小板数，机制可能与抑制了T-bet和GATA-3的表达失常导致的Th1细胞极化有关。
목적：탐토간충질간세포（ MSC）대원발면역성혈소판감소증（ ITP）소서혈소판수목적영향급기궤제。방법채용복강주사대서항소서CD41항체（ MWReg30단항）적방법유도BALB/c소서산생ITP후，취20지작위실험조，20지작위대조조。지후통과서미정맥급실험조소서주사MSC 2×107개/지。주사후5 d、7 d、14 d，측정실험조여대조조소서외주혈혈소판수；14 d시응용역전록취합매련반응（ RT-PCR）검측소서외주혈단개핵세포（ PBMC）중전록인자T-bet화GATA-3 mRNA적표체；응용매련면역법측정Th1류세포인자IFN-γ、IL-2급Th2류세포인자IL-4、IL-10적수평。결과7 d、14 d후，주사MSC실험조소서혈소판치[（588．0±81．6）×109/L、（623．0±78．9）×109/L]명현고우미주사MSC대조조[（317．0±90．1）×109/L、（288．0±87．8）×109/L]（P＜0．05）；14 d시，실험조PBMC중T-bet mRNA표체수평저우대조조[（0．04±0．03） vs （0．27±0．05）]（P＜0．05）；GATA-3 mRNA적표체수평고우대조조[（0．14±0．04） vs （0．07±0．05）]（P＜0．05）；실험조Th1류세포인자IFN-γ、IL-2수평[（3．1±1．7） pg/ml、（3．2±2．1） pg/ml]명현저우대조조[（10．3±4．8） pg/ml、（16．3±5．7） pg/ml]（P＜0．05）；실험조외주혈 Th2류세포인자 IL-4、IL-10수평[（88．6±15．2） pg/ml、（38．3±11．8） pg/ml]명현고우대조조[（32．7±5．7） pg/ml、（22．1±3．4） pg/ml]（P＜0．05）。결론 MSC가이유효제승ITP소서적혈소판수，궤제가능여억제료T-bet화GATA-3적표체실상도치적Th1세포겁화유관。
Objective To explore the effects of mesenchymal stem cells ( MSC ) treatment on platelet counts in mice with immune-mediated thrombocytopenia ( ITP) and the possible mechanism .Meth-ods ITP was induced by daily intraperitoneal injection of anti-platelet membrane CD 41 antibody (MWReg30) into BALB/c mice.The mice were then divided into experiment and control groups with 20 mice in each.Each mouse in experimental group was injected with 2×107 mesenchymal stem cells (MSC) through the tail vein .The numbers of blood platelets in mice from two groups were counted on days 5, 7 and 14 after MSC injection .Reverse transcriptase polymerase chain reaction ( RT-PCR) was performed to meas-ure T-bet and GATA-3 gene expression in peripheral blood mononuclear cells ( PBMCs ) at mRNA level on day 14.The levels of IFN-γ, IL-2, IL-4 and IL-10 in serum were detected by ELISA .Results The platelet counts in experimental group were significantly higher than those in control group on days 7 and 14 after MSC injection [(588.0±81.6)×109/L and (623.0±78.9) ×109/L vs.(317.0±90.1) ×109/L and (288.0± 87.8)×109/L ] (P<0.05).On day 14 after MSC injection, the T-bet expression at mRNA level in PBMCs from mice in experimental group was significantly lower than that in control group [(0.04±0.03) vs.(0.27 ±0.05)] (P<0.05), while the GATA-3 expression at mRNA level was higher than those in control group [ (0.14±0.04) vs.(0.07±0.05)] (P<0.05).Compared with control group, the concentrations of Th1 type cytokines such as IFN-γand IL-2 were remarkably down-regulated in experimental group [(3.1±1.7) pg/ml and (3.2±2.1) pg/ml vs.(10.3±4.8) pg/ml and (16.3±5.7) pg/ml](P<0.05), while the con-centrations of Th2 type cytokines such as IL-4 and IL-10 were up-regulated in experimental group [(88.6± 15.2) pg/ml and (38.3±11.8) pg/ml vs.(32.7±5.7) pg/ml and (22.1±3.4) pg/ml ] (P<0.05). Conclusion MSC treatment can effectively increase platelet counts in mice with immune-mediated thrombo-cytopenia, which may be associated with the suppression of Th 1-dominant response mediated by abnormal ex-pression of T-bet and GATA-3.