Effect of Ginsenoside Rgl on Expression of Integrin and Regulation of Signal Transduction Pathway in Osteoblasts Co-cultured with Ti Particles

Abstract

目的:通过研究人参皂甙Rg1对体外钛微粒诱导的成骨细胞整合素通路相关因子的影响,以期为临床运用人参皂甙Rg1防治人工关节无菌性松动提供实验依据。方法:用人参皂甙Rg1与钛微粒共培养成骨细胞,通过ELISA法检测细胞液中的FN,Col-Ⅰ及VN含量,Western Blot检测MG-63细胞FAK和FAK(pTyr397)蛋白的含量,qPCR法检测MG-63细胞内FAK,Integrinαv及Integrinβ1 mRNA的表达。结果:正常组细胞较大,视野内细胞数最多,实验组细胞数量次之,对照组最少且形态较小,钛微粒周围无细胞生长,并可见凋亡细胞。实验组细胞液中FN,Col-Ⅰ及VN的表达较对照组有明显增高,细胞mRNA与蛋白中FAK,FAK(pTyr397),Integrinαv及Integrinβ明显高于对照组。结论:人参皂甙Rg1可能通过直接或间接作用于MG-63Integrin及其相关信号因子,使与钛微粒共培养的MG-63细胞被抑制的FAK磷酸化再次增强,从而VN和FN功能增强,加速细胞的分裂增殖。Objective:To study the effect of Ginsenoside Rgl on integrin pathway-related factors of osteoblasts induced by titanium particles in vitro, so as to provide experimental basis for the clinical application of Ginsenoside Rgl in the prevention and treatment of aseptic loosening of artificial joints. Methods:The osteoblasts were co-cultured with Ginsenoside Rgl and Ti particles. The con- tent of FN,Col-I and VN in cytoplasm was detected by ELISA method,the content of FAK and FAK(pTyr397) protein in MG- 63 cells was detected by western blot, the expression of FAK, integrinav and integrint31 mRNA in MCJ-63 cells was detected by qPCR method. Results:The cells in the normal group were larger, and the number of the cells within the view was the highest. The number of cells in the experimental group was the second, while the control group was the smallest and the morphology was smal- ler. There was no cell growth around Ti particles, and apoptotic cells were found near the Ti particles. The expression of FN,Col- I and VN in the cytoplasm in the experimental group was increased significantly than that in the control group. The expression of mRNA,FAK,FAK(pTyr397) ,Integrinav and Integrin]3 protein was significantly higher when compared with the control groups. Conclusion:Ginsenoside Rgl may enhance the phosphorylation of FAK again, which is inhibited by MC,-63 cells co-cultured with Ti particles,by direct or indirect acting on MG-63 Integrin and its related signaling factors,so that the function of VN and FN is enhanced to accelerate cell division and proliferation.国家自然科学基金(81302986); 福建省卫生厅中医课题(wzgs201308

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