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背景:葛根素属异黄酮类物质,研究发现其具有减轻酒精毒性和酒精戒断症状的作用,此外还可以抗氧化、清除组织及细胞产生的氧自由基、减轻脂质过氧化对细胞的损害。目的:探讨葛根素对酒精导致的人骨髓间充质干细胞成脂分化的抑制作用。设计、时间及地点:细胞学体外观察,于2008-10在郑州大学基础医学院生物化学与分子生物学教研室实验室完成。材料:骨髓取自健康成人志愿者,由郑州大学第一附属医院骨科提供。葛根素标准品由中国药品生物制品检定所提供。方法:采用梯度离心法分离培养人骨髓间充质干细胞,取传2代细胞,随机分为3组:空白对照组加入含体积分数为10%胎牛血清的DMEM培养基;模型组每天加入乙醇0.09mol/L;实验组每天加入酒精0.09mol/L,每两三天更换培养液时加入葛根素10-6mol/L。各组细胞培养7d。主要观察指标:采用RT-PCR法检测PPAR-γ2基因的表达,油红“O”染色进行脂肪细胞计数,测定细胞内碱性磷酸酶活性。结果:模型组PPAR-γ2mRNA呈高表达,明显高于实验组和空白对照组(P<0.05);实验组PPAR-γ2mRNA表达略高于对照组,但差异无显著性意义(P>0.05)。模型组脂肪细胞最多,细胞内脂滴大而丰富;实验组脂肪细胞较模型组显著减少(P<0.05);空白对照组脂肪细胞极少。与空白对照组比较,模型组细胞内碱性磷酸酶活性明显降低(P<0.05);与模型组比较,实验组细胞内碱性磷酸酶活性明显升高(P<0.05)。结论:葛根素能抑制酒精导致的骨髓间充质干细胞向脂肪细胞分化。
BACKGROUND: Puerarin is an isoflavone substance. It has been found that puerarin has the effects of reducing alcohol toxicity and alcohol withdrawal symptoms. In addition, it can also oxidize, scavenge oxygen free radicals produced by tissues and cells, and reduce the damage of lipid peroxidation to cells. . Objective: To investigate the inhibitory effect of puerarin on alcohol-induced adipogenic differentiation of human bone marrow mesenchymal stem cells. DESIGN, TIME AND SETTING: The in vitro observation of cytology was performed at the Laboratory of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Zhengzhou University from October to October 2008. MATERIALS: Bone marrow was obtained from healthy adult volunteers and was provided by Department of Orthopaedics, First Affiliated Hospital of Zhengzhou University. The puerarin standard is provided by the China National Institute for the Control of Pharmaceutical and Biological Products. METHODS: Human bone marrow mesenchymal stem cells were isolated and cultured by gradient centrifugation and passaged for 2 passages. They were randomly divided into 3 groups. The control group was supplemented with DMEM medium containing 10% fetal bovine serum; the model group was treated with ethanol daily. 0.09mol/L; The experimental group added alcohol 0.09mol/L every day. Puerarin 10-6mol/L was added every two or three days when the medium was changed. Each group of cells was cultured for 7 days. MAIN OUTCOME MEASURES: RT-PCR was used to detect the expression of PPAR-γ2 gene, oil red “O” staining was used to count adipocytes, and the activity of alkaline phosphatase in cells was determined. Results: The expression of PPAR-γ2 mRNA in the model group was significantly higher than that in the experimental group and the blank control group (P<0.05). The expression of PPAR-γ2 mRNA in the experimental group was slightly higher than that in the control group, but there was no significant difference (P>0.05). The fat cells in the model group were the most and the intracellular lipid droplets were large and abundant; the adipocytes in the experimental group were significantly decreased compared with the model group (P<0.05); the adipocytes in the blank control group were few. Compared with the blank control group, the alkaline phosphatase activity in the model group was significantly decreased (P<0.05). Compared with the model group, the activity of alkaline phosphatase in the experimental group was significantly higher (P<0.05). CONCLUSION: Puerarin can inhibit the differentiation of bone marrow mesenchymal stem cells into adipocytes caused by alcohol.