摘要
目的研究再生障碍性贫血(以下简称再障)患儿骨髓间充质干细胞(MSC)体外对淋巴细胞的免疫抑制能力。方法2006年4月至2007年10月重庆医科大学附属儿童医院血液科采集24例再障患儿和11例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;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;采用逆转录PCR(RT-PCR)的方法检测骨髓MSC细胞内转化生长因子-β1(TGF-β1)和肝细胞生长因子(HGF) mRNA的表达水平;采用酶联免疫吸附实验(ELISA)检测骨髓MSC分泌的TGF-β1和HGF的浓度。结果(1)再障患儿骨髓MSC呈典型的成纤维样细胞形态,成纤维集落形成单位(CFU-F)计数显著低于对照组,差异有统计学意义(t=-2.74,P<0.05);(2)再障患儿骨髓MSC免疫抑制率显著低于对照组,差异有统计学意义(t=-3.48,P<0.01);(3)再障患儿骨髓MSC细胞内TGF-β1 mRNA表达水平显著低于对照组,差异有统计学意义(t=-3.34,P<0.01);HGF mRNA表达水平显著低于对照组,差异有统计学意义(t=-2.54,P<0.05);(4)再障患儿骨髓MSC培养上清中TGF-β1浓度显著低于对照组,差异有统计学意义(t=-2.48,P<0.05);再障患儿骨髓MSC培养上清中HGF浓度显著低于对照组,差异有统计学意义(t=-2.46,P<0.05)。结论再障患儿骨髓MSC的免疫抑制能力较对照组显著降低,其发生机制可能与再障患儿骨髓MSC增殖能力减低、细胞表达和分泌TGF-β1、HGF减少有关。
Objective To study the immunosuppressive activity of MSC from controls or patients with AA in vitro. Methods Bone marrow samples were collected from patients with AA at diagnosis[n = 24, 12 from severe aplastic ane- mia（SAA）, 12 from chronic aplastic anemia（CAA）]and the patients whose findings of bone marrow aspiration were nor- mal as the controls （n = 11 ）. MSCs from bone marrow samples were isolated, cultured and expanded. Morphology, pro- liferative 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 and transforming growth faetor-[31 mRNA expression were detected by RT-PCR. HGF and TGF-β1 concentrations of MSC were tested by enzyme-linked immunosorbent assay （ELISA）. Results （1） The culture cells from the patients with AA presented a typical fibroblast-like morphology. The number of CFU-F from the patients with AA （15.7±5.78） was fewer than that from the controls （22.33±5.33）, t = -2.74, P 〈 0.05. （2） The inhibition ratio of MSC from patients with AA[（49.78± 20.36）% ]was significantly lower than that from the controls[（76.42±8.80）% ], t = -3.48, P 〈 0.01. （3） The level of TGF-β1 mRNA expression in MSC from patients with AA was significantly lower than the controls, t = -3.34,P 〈 0.01, and the level of HGF mRNA expression in MSC from patients with AA was significantly lower than the controls, t =-2.54, P 〈 0.05. （4）tThe concentration of TGF-β1 in MSC supernatants from the patients with AA [（415.09±108.10） pg/ml] was significantly lower than the con- trols [（632.40 ± 126.90） pg/ml], t =-2.48, P 〈 0.05 ; the concentration of HGF in MSC supernatants from the patients with AA [（340.72±40.44） pg/ml]was significantly lower than that from the controls [（521.56±36.72）pg/ml], t =-2.46, P 〈 0.05. All these differences were of statistical significance. Conclusion The suppress
出处
《中国实用儿科杂志》
CSCD
北大核心
2009年第10期786-790,共5页
Chinese Journal of Practical Pediatrics
