Simvastatin Affects The Proliferation Of MC3T3-E1 Cells Via ERK1/2 And P38MAPK Signaling Pathways

Osteoporosis is a systemic bone disease characterized by changes in bone density,accelerated bone loss,and reduced and narrowed bone trabeculae.Simvastatin is a natural medicine that can significantly enhance bone formation in addition to its lipid-lowering effect.Its side effects are lower than other therapeutic drugs.In order to explore the effect of simvastatin on the proliferation of MC3T3-E1 cells,and to understand its relationship with the concentration of simvastatin and the time of action;by inhibiting the activity of ERK1/2 and p38 in the MAPK signaling pathway,to explore whether simvastatin passes through ERK1/2,p38MAPK signaling pathway affects the expression of proliferation markers and protein of MC3T3-E1 cells.By adding simvastatin with a concentration of 0mol/,10^-9mol/L,10^-8mol/L,10^-7mol/L,10^-6mol/L,10^-5mol/L into a 96-well cell culture plate,culture for 24h Afterwards,CCK-8 and p NPP kits were used to detect the proliferation activity of MC3T3-E1 cells and the activity of alkaline phosphatase（Alkaline phosphatase,ALP）,and the optimal concentration of simvastatin was selected;when MC3T3-E1cells were confluent Add the optimum concentration of simvastatin to the 96-well plate,set four drug action times（24h,36h,48h,and 60h）.At different time periods,use the CCK-8 kit to determine the proliferation activity of MC3T3-E1 cells Select the optimal time of action of simvastatin;the results show that simvastatin at a concentration of 10^-5mol/L enhances the proliferation and ALP activity of MC3T3-E1cells more significantly;when the action time of simvastatin is 36h,The proliferation effect of MC3T3-E1 cells is more significant.Divide the experiment into 4experimental groups,choose simvastatin with a concentration of 10^-5mol/L,and the action time for 36h,using CCK8 method,p NPP method,enzyme-linked immunosorbent technique（ELISA）,Western blotting（Western blotting）The method was used to detect the ability of cell proliferation,ALP activity,the ability to secrete type I collagen（COL-I）,and the expression levels of p-ERK1/2 and p-p38 in each group.The results showed that compared with the simvastatin group in the blank group,the cell proliferation,ALP activity,and p-ERK1/2 and p-p38 protein expression levels in the simvastatin group were significantly increased,and the secretion of COL-I increased,that is,simvastatin Group can improve MC3T3-E1 cell proliferation,ALP activity,COL-I secretion and p-ERK1/2,p-p38 protein expression levels;Simvastatin+p38 inhibitor group（SIM+SB203580）and Simvastatin+ERK1/2 inhibitor group（SIM+SCH772984）Compared with simvastatin group,after inhibitors of ERK1/2 and p38MAPK signaling pathway act on MC3T3-E1 cells,the cell proliferation effect is significantly weakened,ALP activity and COL-I secretion and p-ERK1/2,p-p38 protein expression ability was significantly reduced.The above results show that 10^-9-10^-5mol/L simvastatin has the effect of promoting the proliferation of MC3T3-E1 cells,and the optimal simvastatin concentration for promoting the proliferation of MC3T3-E1 cells is 10^-5mol/L,which is the best The action time is 36 hours;Simvastatin can enhance the proliferation of MC3T3-E1 cells through the ERK1/2 and p38MAPK signal transduction pathways,increase the ALP activity and the ability to secrete type I collagen,thereby playing a positive role in the proliferation of MC3T3-E1 cells Regulation effect.Provide a reliable theoretical basis for the clinical research of osteoporosis treatment drugs.