CAJ | 학술논문

目的探讨输注间充质干细胞(MSC)对小鼠原发性免疫性血小板减少症(ITP)的治疗效果及其机制.方法采用腹腔注射大鼠抗小鼠CD41抗体的方法诱导建立雌性BALB/c小鼠ITP模型,取建模成功小鼠20只作为实验组,经尾静脉注射雄性BALB/c小鼠的MSC 2×107/个；另取20只建模成功小鼠作为对照组,不注射MSC.注射MSC 14 d后,取两组小鼠外周血,测定血小板数,应用流式细胞术检测外周血髓系树突细胞(mDC,Lin-HLA-DR+ CD11c+细胞)绝对数及其表面共刺激分子CD80和CD86的表达水平,用酶联免疫吸附试验测定TH1细胞因子γ干扰素(INF-γ)、白细胞介素2(IL-2)及TH2细胞因子IL-4、IL-10的水平.结果注射MSC 14 d后,实验组小鼠血小板数量为(623±78.9)×109/L,明显高于对照组的(288±87.8)×109/L(P＞0.05).实验组mDC绝对数量为(16.24±9.97)/ml,明显低于对照组的(29.32±13.21)/ml (P＜0.05);实验组mDC表面的CD80和CD86的表达分别为(2.31±1.26)％和(46.25±20.21)％,明显低于对照组的(6.27±3.31)％和(62.21±16.41)％(P＜0.05).实验组外周血INF-γ和IL-2水平分别为(3.1±1.7)和(3.2±2.1)pg/ml,明显低于对照组的(10.3±4.8)和(16.3±5.7)pg/ml(P＜0.05)；IL4和IL-10水平分别为(88.6±15.2)和(38.3±11.8)pg/ml,明显高于对照组的(32.7±5.7)和(22.1±3.4) pg/ml(P＜0.05).结论 MSC可以有效治疗小鼠ITP,其机制与MSC抑制小鼠DC活化导致TH1细胞的极化有关.
목적 탐토수주간충질간세포(MSC)대소서원발성면역성혈소판감소증(ITP)적치료효과급기궤제.방법 채용복강주사대서항소서CD41항체적방법유도건립자성BALB/c소서ITP모형,취건모성공소서20지작위실험조,경미정맥주사웅성BALB/c소서적MSC 2×107/개；령취20지건모성공소서작위대조조,불주사MSC.주사MSC 14 d후,취량조소서외주혈,측정혈소판수,응용류식세포술검측외주혈수계수돌세포(mDC,Lin-HLA-DR+ CD11c+세포)절대수급기표면공자격분자CD80화CD86적표체수평,용매련면역흡부시험측정TH1세포인자γ간우소(INF-γ)、백세포개소2(IL-2)급TH2세포인자IL-4、IL-10적수평.결과 주사MSC 14 d후,실험조소서혈소판수량위(623±78.9)×109/L,명현고우대조조적(288±87.8)×109/L(P＞0.05).실험조mDC절대수량위(16.24±9.97)/ml,명현저우대조조적(29.32±13.21)/ml (P＜0.05);실험조mDC표면적CD80화CD86적표체분별위(2.31±1.26)％화(46.25±20.21)％,명현저우대조조적(6.27±3.31)％화(62.21±16.41)％(P＜0.05).실험조외주혈INF-γ화IL-2수평분별위(3.1±1.7)화(3.2±2.1)pg/ml,명현저우대조조적(10.3±4.8)화(16.3±5.7)pg/ml(P＜0.05)；IL4화IL-10수평분별위(88.6±15.2)화(38.3±11.8)pg/ml,명현고우대조조적(32.7±5.7)화(22.1±3.4) pg/ml(P＜0.05).결론 MSC가이유효치료소서ITP,기궤제여MSC억제소서DC활화도치TH1세포적겁화유관.
Objective To investigate the efficiency of mesenchymal stem cells (MSCs) in the mice with primary immune thrombocytopenia (ITP) and its mechanism.Method ITP was induced by daily intraperitoneal injection of anti-platelet membrane CD41 antibody into BALB/c mice,then 20 cases were enrolled as observation group and-20 cases as control group at the same time.2 × 107 MSCs were injected into each mouse in observation group through the tail vein.After 14 days,the number of blood platelets was counted in two groups,and the absolute number of myeloid DC (Lin-HLA-DR+CD11c+ cells) and the expression of CD80 and CD86 on mDC of ITP patients were anylyzed by using flow cytometry.The levels of INF-γ,IL-2,IL-4 and IL-10 were detected by using ELASA.Results The number of blood platelets in observation group [(623± 78.9) × 1 09/L] was significantly greater than that in control group [(288 ± 87.8) × 109/L] (P＜0.05).The absolute number of myeloid DC in observation group [(16.24± 9.97)/ml] was significantly less than that in control group [(29.32±13.21)/ml] (P＜0.05).The expression of CD80 and CD86 on mDC in observation group [(2.31 ±1.26)％ and (46.25 ± 20.21)％ respectively] were significantly lower than that in control group [(6.27 ± 3.31) ％ and (62.21 ± 16.41) ％ respectively].The concentrations of Th1 type cytokins INF-γ and IL 2 in observation group [(3.1 ± 1.7) pg/ml and (3.2 ± 2.1) pg/ml,respectively] were significantly lower than those in control group [(10.3 ± 4.8) pg/ml and (16.3 ± 5.7) pg/ml respectively] (P＜0.05),but those of Th2 type cytokins IL-4 and IL-10 in observation group [(88.6 ± 15.2) pg/ml and (38.3 ± 11.8) pg/ml respectively] were significantly higher than those in control group [(32.7 ± 5.7) pg/ml and (22.1 ± 3.4) pg/ml respectively] (P＜0.05).Conclusion MSCs are associated with a favorable efficiency in mice with primary ITP,which was related with the suppression of Th1-dominant response due to the enhanced function of DC.