Study on promotion effect of Qushi Jiangu Formulaon osteogenic differentiation of MC3T3-E1 cells
Objective To explore the mechanism of action on MC3T3-E1 cells.Methods The solution were mixed with different concentrations(0.005 mg/mL,0.025 mg/mL,0.05 mg/mL,0.1 mg/mL,0.2 mg/mL,0.4 mg/mL),and the cells were intervened with different concentrations.After 48 hours of intervention,the cell proliferation rate was detected by Cell Counting Kit-8(CCK-8)method,and the best drug concen-tration was screened.MC3T3-E1 cells were made into cell suspension and randomly divided into blank control,dexamethasone,low dose drug(D),medium dose drug(Z),high dose drug(G)and seeded to a six-well plate.The mold was made after the cell adhesion reached 80%(no drug was added in the blank group;DEX intervention with DEX;D,Z and G intervention with corresponding concentration of bone prescription + DEX)for 48 hours,it replaced with osteogenic induction medium for culture.After 15 days of induction,the alkaline phosphatase(ALP)content and the expression levels of RUNX 2 and OSX protein were measured in each group.Results Compared with the results of CCK-8 experiment,the first three survival concentrations were the drug was selected as 0.1 mg/mL(low concentration),0.2 mg/mL(middle concentration)and 0.4 mg/mL(high concentration).Compared with control group,the ALP ac-tivity in the dexamethasone group(P<0.05),the low-dose group,the middle-dose group and the high-dose group was decreased significantly ALP,there was no significant difference between low-dose and mid-dle-dose groups(P>0.05),but the ALP activity in high-dose groups were increased significantly(P<0.05).Compared with the control group,the expression of RUNX2 and OSX protein in the middle-dose group,low-dose group and dexamethasone group was decreased significantly ALP,there was no signifi-cant difference in the expression of OSX and RUNX2 protein between the high-dose group and the control group(P<0.05),but there was significant difference in the expression of OSX and RUNX2 protein be-tween the high-dose group(P<0.05).Conclusion Through the study,it can be found that the bone pre-scription can promote the osteogenic differentiation ability of MC3T3-E1 cells.Meanwhile,it can be in-ferred that it may regulate bone formation related proteins such as RUNX 2,OSX to correct the balance between bone formation and bone resorption and achieve the treatment of osteoporosis.
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