辛伐他汀对骨髓间充质干细胞增殖及分泌功能影响的研究
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江苏省卫生厅科技发展项目(H200508)


Influence of simvastatin on the proliferation and paracrine functions of bone marrow-derived mesenchymal stem cells
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    摘要:

    目的:细胞水平观察不同浓度的辛伐他汀对骨髓间充质干细胞(bone marrow-drived mesenchymal stem cells,BMSCs)的增殖及旁分泌功能的影响并探讨其可能的机制-方法:采用全骨髓贴壁培养法培养SD大鼠的骨髓间充质干细胞,取3代对数生长期细胞,无血清培养12 h,不同剂量的辛伐他汀(0-0.001-0.010-0.100-1.000 -滋mol/L)与MSCs共培养24 h,采用CCK-8试剂盒检测各浓度组BMSCs增殖情况,通过半定量RT-PCR检测不同浓度的辛伐他汀诱导BMSCs中血管内皮生长因子(vascular endothelial growth factor,VEGF)和肝细胞生长因子(hepatocyte growth factor,HGF)的mRNA表达-采用Western blot检测PAkt-AKt的表达-结果:辛伐他汀在一定浓度范围内(0.001~0.100 -滋mol/L)显著提高了BMSCs增殖和VEGF和HGF的mRNA表达水平(与对照组比较,P < 0.01)-辛伐他汀浓度在0.010 -滋mol/L时对BMSCs的增殖和分泌功能的影响最为显著,随着药物浓度的加大,BMSCs的上述功能呈下降趋势-与对照组比较,1.000 -滋mol/L辛伐他汀对BMSCs无显著的促增殖作用,但却明显上调了VEGF和HGF mRNA的表达-0.010 -滋mol/L辛伐他汀组pAKt表达及pAKt/AKt比值显著高于对照组(P < 0.01)-结论:辛伐他汀在一定浓度范围内对骨髓间充质干细胞的增殖及分泌功能有促进作用,其机制可能与Akt信号通路有关-

    Abstract:

    Objective:To investigate the effect of simvastatin at different concentrations on the proliferation and paracrine functions of BMSCs in vitro and explore its possible mechanism. Methods:BMSCs were isolated from Sprague-Dawley rats and expanded by whole bone marrow adherence culture method. Cells of 3 passage,which were at logarithmic phase,were used in experiment. BMSCs were cultured in serum-free medium for 12 hours,then incubated with various concentrations of simvastatin for 24 hours. The influences of simvastatin on BMSCs’ proliferation were assayed with CCK-8 assay,and their mRNA expressions of vascular endothelial growth factor(VEGF) and Hepatocyte growth factor(HGF) were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Besides,Western-blot used to investigate the pAkt and Akt expressions in BMSCs. Results:Simvastatin in a certain range of concentration (0.001 μmol/L ~0.100 μmol/L) significantly increased proliferation and paracrine (VEGF and HGF) functions of BMSCs in vitro(P < 0.01). Simvastatin at 0.01 μmol/L concentration reached the maximum effects on the proliferation and paracrine functions of BMSCs. While the concentrations of simvastatin were more than 0.01 μmol/L,the improvement effects showed a tendency to decline. Simvastatin at 1.000 μmol/L concentration had no significant effect on promoting proliferation of BMSCs,but significantly increased the VEGF and HGF mRNA expression compared with that of the control group. Both pAKt expression and pAKt/AKt ratio in BMSCs were significantly higher than those in the control group after 0.01 μmol/L simvastatin treatment(P < 0.01). Conclusion:Simvastatin in a certain concentration range could improve BMSCs’ proliferation and paracrine capabilities,and the Akt pathway might be involved in the mechanism.

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陈建梅,张馥敏,王连生,姚永伟,赵〓强,马文珠.辛伐他汀对骨髓间充质干细胞增殖及分泌功能影响的研究[J].南京医科大学学报(自然科学版),2010,(1):20-24

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  • 收稿日期:2009-06-15
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