The Study Of The Effects Of Gypenosides On Proliferation And Differentiation Of Osteoblasts Based On Endoplasmic Reticulum Stress And Wnt/β-catenin Signaling Pathway

ObjectiveTo investigate the effect of gypenosides on osteoblasts from the perspective of Wnt/β-catenin pathway and endoplasmic reticulum stress.Methods1.Three SD rats born 24 hours after birth were selected.After dislocation execution,osteoblasts were extracted,cultured,transcribed and identified.2.The fourth generation(P4)of rat osteoblasts were prepared and cultured in medium containing gypenosides of gypenosides at different concentrations.The proliferation of osteoblasts was determined by CCK-8 method under the intervention of different drug gradients.3.The osteogenic capability of osteoblasts(P4)was determined by Alkaline phosphatase activity(ALP)staining with different drug gradient concentrations,and the effect of gypenosides on the differentiation of osteoblasts was detected at different stages.4.Osteoblasts were divided into control group and experimental group.The optimal drug concentration was selected according to the CCK-8 and ALP staining results.mRNA expressions of Wnt2,β-catenin,BMP2,GRP78/BIP,CHOPP,and PDI were detected by qRT-PCR.Expressions of Wnt2,β-catenin,BMP2,GRP78/BIP,CHOP,and PDI were detected by Western blot.Results1.The proliferation rate of osteoblasts was also different under different drug concentration medium intervention.When the drug concentration was 80 g/ml and the intervention time was 5 days,the proliferation rate was the highest.2.The positive rate of ALP staining of osteoblasts which has a concentration of 80 μg/ml was the higher than other groups.3.Compared with the control group,the mRNA levels of Wnt2,β-catenin,GRP78/BIP,PD I in the experimental group were up-regulated with statistical significance(P<0.05);the expressions of CHOP were also increased without statistical significance(P>0.05),while the expression of BMP2 was slightly decreased with no statistical significance(P>0.05).4.Compared with the control group,the expression of Wnt2,β-catenin,BMP2,PDI increased,with statistically significant difference(P<0.05),while the expression of CHOP also increased,the expression of GRP78/BIP decreased without statistically significant difference(P>0.05).Conclusion1.Gypenosides can promote the proliferation and osteogenic differentiation of osteoblasts.2.Gypenosides of gypenodium pentaphyllum can regulate the Wnt/attenual-catenin signaling pathway in osteoblasts,promote the expression of molecular chaperones like GRP78/BIP and PDI,it can help improving the correct folding rate of proteins in endoplasmic reticulum,and have the potential of regulation of endoplasmic reticulum stress.