阵发性睡眠性血红蛋白尿症患者基质研究初探

目的研究与造血密切相关的基质在阵发性睡眠性血红蛋白尿症(PNH)患者中是否存在异常.方法对7例患者骨髓进行成纤维细胞集落(CFU-F)的培养,观察其形态及数量是否存在异常;将分选后的CD34+正常造血干/祖细胞分别加入含有相同数量的患者及正常对照基质的培养基中,观察其扩增及形成集落的情况;用半定量RT-PCR的方法,以自身β-肌动蛋白作为内参对照,检测患者肿瘤坏死因子(TNF)-α、白细胞介素6(IL-6)表达量与正常人是否不同.结果 (1)PNH患者骨髓CFU-F数量(13.0个±1.8个)类似正常人(10.7个±3.7个,P>0.05).(2)PNH患者的骨髓基质细胞对正常人造血干/祖细胞支持能力与正常人相仿.液体培养细胞数为(4.21±1.05)个×104 vs(4.76±1.04)个×104;半固体培养粒细胞集落10.80个±5.40个 vs 1.50个±0.23个;红细胞集落形成单位4.50个±3.51个 vs 0.83个±0.12个,P>0.05).(3)PNH患者骨髓基质细胞的TNF-α(以细胞因子与自身β-肌动蛋白电泳条带强度的比值表示0.55±0.12)和IL-6(0.62±0.13) mRNA表达量与正常人(0.48±0.08、0.68±0.10)相似(P>0.05).结论患者的骨髓成纤维细胞集落形成能力是正常的.患者基质细胞对正常人造血干/祖细胞的支持能力及其TNF-α、IL-6的mRNA表达基本正常.

Study on the stroma of patients with paroxysmal nocturnal hematoglobinuria

OBJECTIVE: To investigate the relationship between the stroma of bome marrow and the abnormality of hematopoiesis in patients with paroxysmal nocturnal hematoglobinuria (PNH). METHODS: The bone marrow stromata from PNH patients was compared with those from normal volunteers in aspects of their CFU-F formation and hematopoietic sustentation for normal CD34+ cells. The stromal IL-6 and TNF-alpha mRNA expression in patients and normal volunteers was also measured by RT-PCR(self beta-actin as contrast). RESULTS: (1) The CFU-F formation ability in PNH patients was very similar to that in normal volunteers (P > 0.05). (2) The bone marrow stromata from patients with PNH had the same hematopoietic maintenance compared to those from normal controls (P > 0.05). (3) The stomal IL-6 and TNF-alpha mRNA expression was similar in PNH patients and in normal volunteers (P > 0.05). CONCLUSION: The CFU-F formation capacity of bone marrow fibroblast was normal in PNH patients. The stromata in patients with PNH had normal hematopoietic maintainence to the normal CD34+ cells. The mRNA expression of IL-6 and TNF-alpha in PNH stromata was almost normal.