CAJ | 학술논문

目的探讨输注同种骨髓间充质干细胞(MSC)对免疫性血小板减少症(ITP)小鼠血小板数及外周血调节性T淋巴细胞(Treg细胞)的影响及其机制.方法取雌性BALB/c小鼠,每天经腹腔注射含2μg大鼠抗小鼠CD41单克隆抗体的磷酸盐缓冲溶液(PBS)200μl诱导建立小鼠ITP模型.取雄性BALB/c小鼠,获取MSC.然后经ITP小鼠尾静脉注射不同剂量MSC.注射后第5、7和14天,检测各组小鼠的血小板数,采用流式细胞术检测外周血CD4+ CD25high CD127lowTreg细胞的比例,并与正常BALB/c小鼠进行比较.结果输注大鼠抗小鼠CD41单抗后24 h,小鼠血小板数急剧下降至谷值水平左右(约为正常水平的1/4),此时小鼠ITP模型诱导成功.注射MSC后,ITP小鼠的血小板数逐渐升高,至注射7d后,明显高于未注射MSC的ITP小鼠(P＜0.05),且注射MSC剂量越高,血小板数升高的程度越大.建模成功后,ITP小鼠的Treg细胞数明显低于正常小鼠(P＜0.05);注射MSC后,ITP小鼠外周血Treg细胞比例升高,且注射MSC剂量越高,作用增强,但仍未达到正常小鼠的水平.结论输注同种MSC可以提高ITP小鼠的血小板数,其机制与上调Treg细胞的水平有关.
목적 탐토수주동충골수간충질간세포(MSC)대면역성혈소판감소증(ITP)소서혈소판수급외주혈조절성T림파세포(Treg세포)적영향급기궤제.방법 취자성BALB/c소서,매천경복강주사함2μg대서항소서CD41단극륭항체적린산염완충용액(PBS)200μl유도건립소서ITP모형.취웅성BALB/c소서,획취MSC.연후경ITP소서미정맥주사불동제량MSC.주사후제5、7화14천,검측각조소서적혈소판수,채용류식세포술검측외주혈CD4+ CD25high CD127lowTreg세포적비례,병여정상BALB/c소서진행비교.결과 수주대서항소서CD41단항후24 h,소서혈소판수급극하강지곡치수평좌우(약위정상수평적1/4),차시소서ITP모형유도성공.주사MSC후,ITP소서적혈소판수축점승고,지주사7d후,명현고우미주사MSC적ITP소서(P＜0.05),차주사MSC제량월고,혈소판수승고적정도월대.건모성공후,ITP소서적Treg세포수명현저우정상소서(P＜0.05);주사MSC후,ITP소서외주혈Treg세포비례승고,차주사MSC제량월고,작용증강,단잉미체도정상소서적수평.결론 수주동충MSC가이제고ITP소서적혈소판수,기궤제여상조Treg세포적수평유관.
Objective To evaluate the effect of the conspecific bone marrow mesenchymal stem cells (MSCs) infusion on the platelets count and the ratio of CD4+ CD25highCD127low regulatory T (Treg) cells in mice with immune thrombocytopenia and the mechanisms.Method ITP mice models were induced by daily intraperitoneal injection of 200 μL phosphate buffer solution [containing 2 μg rat anti-platelet membrane CD41 antibody (MWReg30)] into female Balb/c mice.MSCs were got from male mice.Then different number of MSCs was injected into ITP mice through the tail veins.After 5,7 and 14 days,the number of blood platelets was counted and the ratio of Treg cells was detected by flow cytometry,and compared with those in the healthy mice.Result Twenty-four h after injection of CD41 antibody,platelet counts were reduced sharply to the lowest point,which was about a quarter of the normal level.Then ITP mice models were induced successfully.Platelet counts were increased after the injection of MSCs.On 7th day after injection of MSCs,the platelet counts were significantly higher than those in control mice,and the greater the degree of injection dosage,the greater the elevated platelets (P＜0.05 for all).The ratio of Treg cells in ITP mice models was significantly lower than in the normal mice.The ratio of peripheral blood Treg cells in ITP mice was increased after injection of MSCs and the higher the dose,the greater the effect (P＜0.05 for all) but did not reach the normal level.Conclusion The conspecific bone marrow MSCs infusion can increase the platelet counts in mice with ITP,which may be related to the increase of CD4+ CD25highCD127low Treg cells.