酒精对骨髓多能干细胞PPARγ及骨钙素mRNA表达的影响

Effect of alcohol on PPARγ and osteocalcin mRNA expression in multipotential stem cell from bone marrow

目的 :观察酒精对骨髓多能干细胞 (D1细胞 )成脂转录因子 (PPARγ)mRNA和成骨基因骨钙素mRNA表达的影响。方法 :以不同浓度 (0 (对照组 ) ,0 .0 9,0 .15 ,0 .2 1mol/L)酒精处理D1细胞 14d ,苏丹ⅠV染色 ,采用计算机图像分析软件确定脂肪细胞百分比 ,采用RT PCR技术检测细胞中PPARγ及骨钙素mRNA表达。结果 :对照组脂肪细胞生成很少 ,酒精处理组脂肪细胞生成显著增多 ,脂肪细胞百分比随酒精浓度增大而增多 (P <0 .0 0 1)。各组与对照组PPARγmRNA表达差异无统计学意义。 0 .0 9,0 .15 ,0 .2 1mol/L酒精组骨钙素mRNA表达降低 ,分别比对照组减少 38%、4 3%和 5 4 % (P <0 .0 0 1)。结论 :酒精能够导致骨髓多能干细胞大量分化为脂肪细胞 ,可能是酒精在脂肪酸旁路代谢下游所起的作用 ,这可能与酒精性骨坏死的发生机制有关。

Aim: To observe the effect of alcohol on PPARγ and osteocalcin mRNA expression in multipotential stem cell from bone marrow named D1 cell. Methods: D1 cells were maintained in culture and treated with 0(control), 0.09, 0.15, and 0.21 mol/L ethanol daily for 14 days. The effect of ethanol on adipogenesis was examined by detecting the accumulation of triglyceride vesicles winthin the cells by phase contrast microscopy and Sudan IV staining. The percentage of adipocytes was calculated by using computer image analysis software. The level of PPARγ and osteocalcin mRNA expression was investigated by RT-PCR. Results: On day 14, there were few adipocytes in the control cells and a large number of adipocytes in the cells treated with ethanol. The percentage of adipocytes in culture increased when the cells were treated with higher concentrations of ethanol, which showed a dose-dependent relation (P < 0.001). The expression of osteocalcin mRNA of treatment with 0.09, 0.15, and 0.21 mol/L ethanol decreased by 38%, 43%, and 54%,respectively, compared with control cells (P< 0.001). While expression of PPARγ mRNA did not change compared mith control cells ( P > 0.05). Conclusion: Alcohol can induce the differentiation of multipotential stem cell from bone marrow (D1) into adipocytes, suggesting that adipogenesis is brought about by the action of ethanol downstream in the fatty acid metabolism pathway, which may be a major factor contributing to alcohol-related osteonecrosis.