Effect Of Peroxisome Proliferator Activated Receptorγ Agonist Rosiglitazone On The Proliferation And Differentiation Of MC3T3-E1Cells In Vitro

objective1. To study the effect of Rosiglitazone(RSG),the agonist of peroxisome proliferatoractivated receptorγ(PPARγ),on the proliferation and differentiation of MC3T3-E1cells in vitro and to explore its related cellular pathway mechanism.2. To observe the effects of different concentrations of glucose on the proliferationand differentiation of primary osteoblasts.Methods1. Buying MC3T3-E1cells at Shanghai Institute of Cell Bank. Treating MC3T3-E1cells with different dose of Rosiglitazone, To observe the effects of differentconcentrations of Rosiglitazone on the MC3T3-E1cells proliferation.ALPstaining and Alizarin red staining. Real—time PCR was used for thedetermination of Runx2、 OB markers ALP and OCN mRNA expression.Western blot was used for determination of Runx2、ALP、β-catenin、ERK1/2and p-ERK1/2protein expression.2. The identification of mouse primary osteoblasts was performed byimmunofluorescence staining of collagen typeⅠand Von Kossa staining.Treatingosteoblasts with different dose of glucose, To observe the effects of differentconcentrations of glucose on the osteoblasts proliferation，ALP activity. Real—time PCR was used for the determination of Runx2、OB markers ALP and OCNmRNA expression.Results1. Compared with DMSO group the experimental group cell proliferation decreased.With the increasing Rosiglitazone concentrations, the ALP staining and theAlizarin red staining in1uM RSG group and5uM RSG group decreased. PCRshowed the mRNA expression of Runx2、ALP and OCN decreased，westernblot showed Runx2、ALP protein expression decreased，With the increasingRosiglitazone concentrations， β-catenin、 ERK1/2and p-ERK1/2proteinexpression decreased too.2. With the increasing glucose concentrations,the osteoblasts cell proliferationdecreased. Compared with5.5mmol／L normal glucose control,the ALP stainingin15.5mmol／L group and25.5mmol／L group decreased;PCR showed themRNA expression of OCN、ALP、Runx2decreased. Conclusions1. Rosiglitazone inhibition of MC3T3-E1cells proliferation and osteogenicdifferentiation, this inhibition may be achieved through the Wnt/beta-catenin andERK1/2pathway.2. High glucose can decrease osteoblasts proliferation and activity, Which is one ofthe key pathogenesis factors of diabetic osteoporosis.