强筋壮骨胶囊调控成骨细胞线粒体自噬影响细胞活力的研究OA

Objective:Study the effects of the serum containing drugs from the Qiangjin Zhuanggu capsule on the mitochondrial autophagy and cell viability of osteoblasts,as mediated by the PINK1/Parkin signaling pathway.Methods:SPF-grade SD rats were selected to prepare blank and drug-containing serum from Qiangjin Zhuanggu capsules.The CCK-8 method was then used to determine the optimal concentration of the drug-containing serum for the follow-up experiment.The MC3T3-E1 Subclone 14 cells were divided into the following groups:blank serum,3-MA,Qiangjin Zhuanggu capsule drug-containing serum+3-MA,and Qiangjin Zhuanggu capsule drug-containing serum.Osteoblast activity was detected using the CCK-8 method.Alkaline phosphatase activity was used to detect the bone formation ability of the cells.The mRNA expressions of OCN,PINK1,and Parkin were detected using PT-PCR.The expressions of OCN,PINK1,and Parkin were detected using Western blot.Results:The Qiangjin Zhuanggu capsule promoted the proliferation and differentiation of osteoblasts.The effect was most obvious at a concentration of 1%,at which point the cell proliferation rate significantly increased(P<0.01).Therefore,1%was selected as the optimal concentration for subsequent experiments.Qiangjin Zhuanggu capsule significantly increased bone cell and alkaline phosphatase activity(P<0.01)as well as the mRNA and protein expressions of OCN,PINK1,and Parkin(P<0.01)compared to the other three groups.The bone cell activity and alkaline phosphatase activity of the 3-MA group and the Qiangjin Zhuanggu capsule drug-containing serum+3-MA group were lower than those of the blank serum group(P<0.01),and the mRNA and protein expressions of OCN,PINK1,and Parkin were lower than those of the blank serum group(P<0.01).Qiangjinzhuangu capsule drug serum+3-MA had better bone cell viability and alkaline phosphatase activity than the 3-MA group(P<0.01),and higher mRNA and protein expressions of OCN,PINK1,and Parkin than the 3-MA group(P<0.01).Conclusion:The Qiangjin Zhuanggu capsule can effectively improve the activity of osteoblasts.The mechanism may be related to the activation of the PINK1/Parkin signaling pathway and the regulation of osteoblast mitochondrial autophagy.