Ghrelin联合三氧化二砷对骨髓间充质干细胞增殖与成骨分化的影响

该文主要探究Ghrelin对三氧化二砷(As2O3)导致的骨髓间充质干细胞(BMSCs)增殖和成骨分化的影响。BMSCs设为对照组、As2O3组、Ghrelin组和联合(As2O3+Ghrelin)组。MTT法检测细胞增殖能力;成骨诱导的第7天和第14天,Real-time PCR及Western blot分别检测成骨相关因子OPN、ALP、RUNX2的mRNA及蛋白表达;第21天,茜素红染色分析钙盐沉积情况。结果显示,细胞增殖能力Ghrelin组>对照组>联合组>As2O3组。与对照组比,As2O3组各因子表达均显著下调(P<0.05),Ghrelin组第14天OPN蛋白表达无显著变化,其余因子均上调(P<0.05);联合组与As2O3组比,第14天OPN基因表达和第7天ALP蛋白表达无显著差异,其余均显著上调(P<0.05)。钙盐沉积:Ghrelin组>对照组>联合组>As2O3组。提示0.5μmol/L As2O3抑制BMSCs增殖和成骨分化,600 ng/mL Ghrelin增强细胞增殖和成骨分化;且Ghrelin能减弱As2O3导致的BMSCs增殖和成骨分化抑制作用。

Effects of Combination of Ghrelin and Arsenic Trioxide on the Proliferation and Osteogenic Differentiation of Bone Marrow Derived Mesenchymal Stem Cells

This work was to investigate the effects of Ghrelin on proliferation and osteogenic differentiation of BMSCs(bone marrow mesenchymal stem cells)induced by As2O3(arsenic trioxide).BMSCs were divided into four groups:control group,As2O3 group,Ghrelin group and As2O3+Ghrelin group.MTT assay was used to detect cell proliferation.Real-time PCR and Western blot were used to detect the levels of OPN,ALP and RUNX2 mRNA and protein in BMSCs of different induction groups at day 7,14 of osteogenesis induction.Alizarin red staining was used to analyze the calcium salt deposition of cells in different induction groups at day 21 of osteogenesis induction.The results showed that the cell proliferation ability of Ghrelin group>control group>combined group>As2O3 group.Compared with the control group,As2O3 group significantly decreased the mRNA and protein expression of osteogenesis-related factors OPN,ALP and RUNX2(P<0.05).There was no significant change in OPN protein expression on the 14th day in Ghrelin group,and the other factors were up-regulated(P<0.05).Compared with As2O3 group,the expression of 14th day of OPN and 7th day of ALP in the combined group had no significant difference,and the other factors were up-regulated(P<0.05).Calcium deposition:Ghrelin group>control group>combined group>As2O3 group.The results showed that 0.5μmol/L As2O3 could inhibit the proliferation and osteogenic differentiation of BMSCs,while 600 ng/mL Ghrelin could promote the proliferation and osteogenic differentiation of BMSCs.And Ghrelin could weaken the inhibition of proliferation and osteogenic differentiation of BMSCs induced by As2O3 to some extent.