| Objective: To study the effect of the medicated serum of Jiangu Erxian Pill,Jingui Shenqi Pill and Liuwei Dihuang Pill on the osteogenic differentiation process of bone marrow mesenchymal stem cells(BMSCs),as well as the effect on alkaline phosphatase(ALP)and Dlx5 expression,and to explore the mechanism of three kidney tonifying formulas in the treatment of osteoporosis.Methods: 40 SD rats were divided into three groups: Jiangu Erxian Pill group,Jingui Shenqi Pill group,Liuwei Dihuang Pill group and blank control group,with 10 rats in each group.75.9 g/kg Jiangu Erxian Pill,89.1 g/kg Jinkui Shenqi Pill,85.8 g/kg Liuwei Dihuang Pill and equal volume of normal saline were gavaged into the stomach respectively,and the serum containing the drug was prepared by gavage twice a day for 7 days.P3 BMSCs were treated with H2O2 to form senescent cells.The aging BMSCs were divided into 6 groups: model group,induction group,Jiangu Erxian Pill containing serum group,Jingui Shenqi Pill containing serum group,Liuwei Dihuang Pill containing serum group,blank serum group(The serum concentration of each group is 10%).At the 4th,8th,12 th and 16 th day of induction,the Dlx5 m RNA and protein expression levels were detected by RT-q PCR and WB methods,and the cell supernatant was collected at the 4th and 8th day to detect ALPactivity.Results: On the 4th and 8th day,compared with the model group,the expression of ALP activity in the induction group and the blank serum group increased significantly(P<0.05).Compared with the induction group and the blank serum group,the expression of ALP activity in the Jiangu Erxian Pill medicated serum group,the Jingui Shenqi Pill medicated serum group and the Liuwei Dihuang Pill medicated serum group was significantly higher(P<0.05).On the 4th and 12 th day,compared with the model group,the expression level of Dlx5 m RNA and protein in the induction group and blank serum group increased(P<0.05);Compared with the induction group and the blank serum group,the expression level of Dlx5 m RNA and protein in the Jiangu Erxian Pill containing serum group,the Jingui Shenqi Pill containing serum group,and the Liuwei Dihuang Pill containing serum group increased(P<0.05).On the 8th and 16 th day,compared with the model group,the expression level of Dlx5 m RNA and protein in the Jiangu Erxian Pill serum group increased(P<0.05).Conclusion:Jiangu Erxian Pill,Jingui Shenqi Pill and Liuwei Dihuang Pill can induce osteogenic differentiation of senile BMSCs and promote the expression of Dlx5 and ALP activity. |
