摘要
目的:研究特发性血小板减少性紫癜(Idiopa thicthrombocytopenic purpura,ITP)患儿骨髓间充质干细胞(Mesenchymal stem cells,MSC)体外的免疫抑制能力。方法:采集ITP患儿和对照儿童的骨髓标本,分离、培养和扩增MSC,观察细胞形态及增殖能力,计数分析骨髓MSC成纤维细胞集落形成单位(Colony forming unit-fibroblast,CFU-F);采用混合淋巴细胞反应检测骨髓MSC体外对淋巴细胞的免疫抑制能力;采用RT-PCR和ELISA方法检测骨髓MSC细胞内转化生长因子-β1(Transforming growth factor-β1,TGF-β1)和肝细胞生长因子(Hepatocyte growth factor,HGF)mRNA的表达水平和细胞培养上清中因子的浓度。结果:(1)ITP患儿的骨髓MSC呈典型的成纤维样细胞形态,CFU-F计数(21.29±5.25)与对照组骨髓MSC的CFU-F计数(22.33±5.33)无显著差异(P>0.05);(2)ITP患儿骨髓MSC免疫抑制率([74.93±8.01)%]与对照组骨髓MSC免疫抑制率([76.42±8.80)%]比较无显著性差异(P>0.05);(3)ITP患儿骨髓MSC细胞内TGF-β1、HGF mRNA的表达水平与对照组比较均无显著性差(P>0.05);(4)ITP患儿骨髓MSC培养上清中TGF-β1的浓度([617.88±81.89)pg/ml]与对照组骨髓MSC培养上清中TGF-β1的浓度([632.40±126.90)pg/ml]比较无显著性差异(P>0.05);ITP患儿骨髓MSC培养上清中HGF浓度([520.20±30.67)pg/ml]与对照组骨髓MSC培养上清中HGF的浓度([521.56±36.72)pg/ml]比较无显著性差异(P>0.05)。结论:ITP患儿骨髓MSC TGF-β1和HGF的表达水平及免疫抑制能力与对照组比较无显著性差异。
Objective： Idiopathic thrombocytopenic purpura（ITP） is characterized by a low platelet count and mucoeutaneous bleeding. The immunity abnormality plays important role in the pathophysiology of idiopathic thrombocytopenic purpura. Mesenchymal stem ceils （MSC） have been shown to possess immunosuppressive activity, preventing lymphocytes activiation. The aim of the study was to compare the immunosuppressive activity of MSC from controls and patients with ITP in vitro. Methods： Bone marrow samples were collected from patients with ITP at diagnosis（ n=19 ）, and the patients whose findings of bone marrow aspiration showed normal were established as the controls（n=11 ）. MSCs from bone marrow samples were isolated, cultured and expanded. Morphology, proliferative activity and colony forming unit-fibroblast （ CFU-F ） were measured and analyzed. MSC were assayed for the activity of lymphocytes suppression by mixed lymphocyte reaction（MLR）. The hepatocyte growth factor,transforming growth factor-β1 mRNA expression was detected by RT-PCR. HGF, TGF- β1 release of MSC was tested by ELISA. Results： （ 1 ） The culture cells from the patients with ITP presented a typical fibroblast-like morphology. The numbers of CFU-F from the patients with ITP （21.29 ± 5.25） showed no difference from the controls （22.33 ± 5.33 ） ,P〉0.05. （2） The inhibition ratio of MSC in patients with ITP [（74.93 ± 8.01 ）% ] showed no difference from that in the controls[（76.42 ± 8.80）% ], P〉 0.05 ; （3） The TGF- β1, HGF mRNA expression in MSC in patients with ITP were similar to those in the controls,P〉O.05 ; （4）The concentration of TGF-β1 in MSC supernatants in the patients with ITP[（617.88 ± 81.89 ）pg/ml] was similar to that in the controls [（632.40± 126.90）pg/ml],P〉0.05;the coneentration of HGF in MSC supernatants in the patients with ITP [（520.20 ± 60.67 ） pg/ml] was similar to that in the controls [（521.56 ±36.72 ）pg/ml], P〉0.05. Conclusion： The suppressiv
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2009年第3期260-264,共5页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(30100095)
