EGCG联合力学刺激对成骨细胞增殖与分化的影响

Effects of epigallocatechin-3-gallate coupled with mechanical stimulation on the proliferation and differentiation of osteoblast

目的探究表没食子儿茶素没食子酸酯(EGCG)联合力学刺激(MS)对成骨细胞增殖分化的诱导作用。方法实验分为对照组、EGCG组、MS组、EGCG+MS组;采用MTT法检测EGCG对成骨细胞的增殖活性;采用茜素红染色法检测EGCG对成骨细胞矿化的影响;采用四点弯曲力学加载装置向细胞施加力学载荷,应用碱性磷酸酶(ALP)测试盒检测ALP活性;采用免疫印迹法检测成骨细胞runt相关转录因子-2(Runx-2)和I型胶原蛋白(Col-I)的表达。结果浓度为30μmol/L的EGCG对成骨细胞的增殖作用最明显,差异有统计学意义(P<0.05);EGCG组的矿化结节数量多于对照组,与单一EGCG组和MS组相比,EGCG+MS组ALP活性明显提高,Runx-2和Col-I蛋白的表达增加(P<0.05)。结论EGCG联合力学刺激增强了成骨细胞ALP活性以及Runx-2和Col-I蛋白的表达,成骨细胞的增殖与分化能力增加。

Objective To investigate the effect of epigallocatechin gallate(EGCG)coupled with mechanical stimulation(MS)on the proliferation and differentiation in osteoblasts.Methods The osteoblasts were divided into control group,EGCG group,MS group,and EGCG+MS group.The proliferation of osteoblasts was detected by MTT assay.The mineralisation of osteoblasts were detected by alizarin red staining method.The mechanical load was applied to the cells by a four-point bending mechanical loading device,and the Alkaline Phosphatase(ALP)Test Kit was used to detect the ALP activity.The protein expressions of runt-related transcription factor-2(Runx-2)and collagen type I(Col-I)in osteoblasts were detected by western blot.Results EGCG at a concentration of 30μmol/L had the most pronounced proliferative effect on osteoblasts,with a statistically significant difference(P<0.05).The number of mineralised nodules in the EGCG group was greater than that in the control group.Compared with the single EGCG or MS groups,the ALP activity,in the EGCG+MS group was significantly increased and the protein expressions of Runx-2 and Col-I in the EGCG+MS group were significantly increased(P<0.05).Conclusion EGCGcoupled mechanical stimulation enhances the ALP activity,up-regulates the expressions of Runx-2 and Col-I proteins in osteoblasts,and promotes osteoblasts′proliferation and differentiation.