HU-308对高浓度糖皮质激素作用下MC3T3-E1成骨细胞生物学性能的影响

Effects of HU-308 on Biological Properties of MC3T3-E1 Osteoblasts at High Glucocorticoids Concentrations

目的:观察HU-308对高浓度糖皮质激素作用下成骨细胞生物学性能的影响。方法:将MC3T3-E1细胞在高浓度糖皮质激素(10-6 mmol/L)进行培养,按HU-308浓度分为0(对照组)、10-10、10-9、10-8、10-7、10-6 mol/L(实验组)6组,培养1、4、7d检测细胞增殖及碱性磷酸酶(alkaline phosphatase,ALP)活性、成骨基因特异性转录因子Runt相关基因2(runt related transcription factor-2,RUNX2)及护骨素(osteoprotegerin,OPG)mRNA的表达,培养24h观察细胞骨架的形态变化。结果:细胞计数试剂盒(cell counting kit-8,CCK-8)结果显示,对照组与实验组相比差异均有统计学意义(P<0.05),培育4d,各实验组比较,药物浓度增加对细胞增殖的促进作用越明显(P<0.05)。ALP活性检测结果显示,实验组与对照组比较差异无统计学意义(P>0.05)。细胞骨架染色随HU-308浓度的增高细胞铺展面积增大且内部框架结构更加清晰。低浓度组Runx2、OPG mRNA表达水平明显上调(P<0.05);而高浓度组Runx2、OPG mRNA的表达受到抑制(P<0.05)。结论:在高浓度糖皮质激素作用下,HU-308可促进MC3T3-E1细胞的粘附、伸展与增殖,且低浓度HU-308可促进其Runx2、OPG mRNA的表达。

Objective: To observe the effects of HU-308 on osteoblast treated with high concentration of glucocorticoid.Methods: MC3T3-E1 cells were cultured in the high concentration of glucocorticoid (10^-6 mmol/L) and then added the HU-308.Cells in concentration of 10^-10,10^-9,10^-8,10^-7,and 10^-6 mol/L of HU-308 were set as the experimental groups,and that with only glucocorticoid was set as the control group.The CCK-8 was used to detect the cell proliferation,ALP activity,and mRNA expression of Runx2 and OPG.The morphology of the cytoskeleton was observed by confocal laser scanning microscope when the MC3T3-E1 cells were cultured for 24h.Results: The results of CCK-8 showed that there was statistically significant between the control group and experimental group (P<0.05).The cell proliferation was effectively promoted in the experimental group (10^-9,10^-8,10^-7 and 10^-6 mol/L) in 4 d (P<0.05).ALP activity detection showed no significant difference between experimental groups and control group (P>0.05).With the concentration of HU-308 increased,cells spreading area increased and the internal frame structure was clearer.The mRNA expressions of Runx2 and OPG were significantly raised at low concentration of HU-308 (P<0.05),however,inhibited at high concentration (P<0.05).Conclusion: HU-308 promoted the proliferation,adhesion,and extension of MC3T3-E1 cells under the action of high concentration of glucocorticoids,and low concentration of HU-308 could promote the expression of Runx2 and OPG mRNA.