| Objective:To investigate the effects of serum containing 3 kinds of bushen prescriptions(Jianggu Erxian Wan,Liuwei Dihuang Wan,Jinkui Shenqi Wan)on aging bone marrow mesenchymal stem cells,marrow mesenchymal stem cells,The effect of osteoblast specific transcription factor(Osterix,Osx)and Osteocalcin(Ocn)expression in the process of osteogenic differentiation of BMSCs.The mechanism of preventing and treating osteoporosis by three bushan prescriptions was studied.Methods:After 1 week of adaptive feeding,40 SPF SD male rats were randomly divided into 4 groups:blank control group(equal volume normal saline),Jiangu Erxian Wan group(75.9 g/kg),Liuwei Dihuang Wan group(85.8 g/kg)and Jinke Shenqi Wan group(89.1 g/kg),with 10 rats in each group.The P3 generation BMSCs with logarithmic growth were established.The oxidative aging BMSCs were treated with 500μmol/L 3 0%H2O2.The metabolic activity of aging BMSCs was detected using cck-8 method.After that,the prepared aging BMSCs were divided into model group,induction group,blank serum group,Jiangu Erxian Wan drug-containing serum group,Jinkui Shenqi Wan drug-containing serum group and Liuwei Dihuang Wan drug-containing serum group.At the 4th,8th,12th and 16th days of induction,Ocn expression level was detected by Elisa,Osx protein expression was detected by WB,and Oxs mRNA relative expression level was detected by RT-PCR.Results:Compared with normal BMSCs,the mean OD of BMSCs treated with H2O2 was significantly decreased(P<0.05).Compared with the model group,Ocn concentration in the induction group and blank serum increased on the 4th,8th,12th and 16th days(P<0.05).Compared with induction group and blank serum group,Ocn concentration in Jiangu Erxian Pill containing serum group and Jinkui Shenqi Pill containing serum group increased on the 4th,8th,12th and 16th days(P<0.05).Compared with induction group,Ocn concentration in Liuwei Dihuang Pill containing serum group was increased on the 4th,8th and 16th days(P<0.05).Compared with blank serum group,Ocn concentration in Liuwei Dihuang Pill containing serum group was increased on day 4,day 12 and day 16(P<0.05).Compared with model group,Osx mRNA and protein expression levels in induction group and blank serum group were significantly increased at day 4,day 8,day 12 and day 16(P<0.05).Compared with induction group and blank serum group,Osx mRNA and protein expression levels in Jiangu Erxian Pill containing serum group and Jinkui Shenqi Pill containing serum group were significantly increased on day 4,day 8,day 12 and day 16(P<0.05).Compared with induction group,Osx mRNA and protein expression levels in Liuwei Dihuang Pill containing serum group were significantly increased on day 4,day 8 and day 16(P<0.05).Compared with blank serum group,Osx mRNA and protein expression levels in Liuwei Dihuang Pill containing serum group were significantly increased on day 4,day 12 and day 16(P<0.05).Conclusion:The three bushen prescriptions can promote the osteogenic differentiation of aging BMSCs,and the effect is better than that of osteogenic medium and blank serum.The osteogenic differentiation ability of BMSCs was directly proportional to the expression of Osx,Ocn and other osteogenic factors.The three kinds of bushen prescriptions can promote the expression of Osx and Ocn,promote bone differentiation of BMSCs,and prevent osteoporosis. |
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