Lin Quan-De, Fang Bai-Jun, Zhou Jian, Zhang Yan-Li, Liu Yang, Wang Chao, Zhao Jun-Mei, Song Yong-Ping
Department of Hematology, Zhengzhou University Affiliated Cancer Hospital; Department of Hematology, Henan Cancer Hospital; Henan Hematology Institute, Zhengzhou 450008,Henan Province,China.
Department of Hematology, Zhengzhou University Affiliated Cancer Hospital; Department of Hematology, Henan Cancer Hospital; Henan Hematology Institute, Zhengzhou 450008, Henan Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2014 Dec;22(6):1667-72. doi: 10.7534/j.issn.1009-2137.2014.06.031.
This study was aimed to explore the regulation of arsenic trioxide (As₂O₃) on imbalance between adipogenic and osteogenic differentiation of BM-MSC from patients with aplastic anemia(AA). The BM-MSC from AA patients were separated and purified, placed into the adipogenic and osteogenic differentiation culture system, then added the As₂O₃, CsA, As₂O₃combined with CsA were added to corresponding differentiation culture system, the concentration of As₂O₃and CsA were set at 0.001 µmol/ml and 2.5 mmol/ml respectively, the cells were divided into As₂O₃group, the CsA group, combined group and control group (no drug). The cell morphological observation, oil red 'O' staining, Von-Kossa staining, and RT-PCR were used to detect corresponding differentiation marks. The results indicated that in respect to adipogenic differentiation, cellular morphology observing and oil red 'O' staining showed that the rate of adipocyte differentiation in As₂O₃group was (18.3 ± 1.9)%, which was lower than the (91.8 ± 2.7)% in the CsA group and (92.1 ± 1.2)% in control group (P < 0.05), there was no significant difference in comparison with (8.3 ± 1.9)% in the combined group (P > 0.05), but the rate of differentiation in CsA group was higher than that in combined group (P < 0.05), and there was no significant difference in comparison wtih control group. RT-PCR showed that the LPL-mRNA expression level in As₂O₃group were significantly lower than that in the CsA group and the control group (P < 0.05), no difference was observed while compared with the combined group (P > 0.05), but the LPL-mRNA expression level in CsA group was significantly higher than that in the combined group (P < 0.05). In terms of osteogenetic differentiation, the calcium deposition in As₂O₃group and combined group was obviously observed while rarely in the CsA group and the control group when detected by the Von-Kossa staining. OST-mRNA expression level in As₂O₃group were higher than that in CsA group and the control group (P < 0.05), while compared with the combined group, there was no significant difference (P > 0.05), but the OST-mRNA expression level in the CsA group was lower than that in the combined group (P < 0.05). It is concluded that As₂O₃can significantly enhance the ability of BM-MSC from AA patients to differentiate into osteoblast, also can inhibit the adipogenic differentiation, in contrast, the CsA can not promote the osteoblast differentiation of BM-MSC from AA patients.
本研究旨在探讨三氧化二砷(As₂O₃)对再生障碍性贫血(AA)患者骨髓间充质干细胞(BM-MSC)成脂与成骨分化失衡的调控作用。分离纯化AA患者的BM-MSC,将其置于成脂和成骨分化培养体系中,然后分别向相应的分化培养体系中添加As₂O₃、环孢素A(CsA)、As₂O₃与CsA的组合,As₂O₃和CsA的浓度分别设定为0.001 μmol/ml和2.5 mmol/ml,细胞分为As₂O₃组、CsA组、联合组和对照组(无药物)。采用细胞形态学观察、油红'O'染色、冯·科萨染色及逆转录-聚合酶链反应(RT-PCR)检测相应的分化标志物。结果表明,在成脂分化方面,细胞形态学观察及油红'O'染色显示,As₂O₃组脂肪细胞分化率为(18.3±1.9)%,低于CsA组的(91.8±2.7)%和对照组的(92.1±1.2)%(P<0.05),与联合组的(8.3±1.9)%比较差异无统计学意义(P>0.05),但CsA组分化率高于联合组(P<0.05),与对照组比较差异无统计学意义。RT-PCR显示,As₂O₃组脂蛋白脂肪酶(LPL)-mRNA表达水平显著低于CsA组和对照组(P<0.05),与联合组比较差异无统计学意义(P>0.05),但CsA组LPL-mRNA表达水平显著高于联合组(P<0.05)。在成骨分化方面,冯·科萨染色检测显示,As₂O₃组和联合组有明显的钙沉积,而CsA组和对照组少见。As₂O₃组骨钙素(OST)-mRNA表达水平高于CsA组和对照组(P<0.05),与联合组比较差异无统计学意义(P>0.05),但CsA组OST-mRNA表达水平低于联合组(P<0.05)。结论:As₂O₃可显著增强AA患者BM-MSC向成骨细胞分化的能力,同时抑制其成脂分化,相反,CsA不能促进AA患者BM-MSC的成骨细胞分化。