Effects Of Ginsenoside Rg1 On Proliferation、Differentiation And Expression Of Inflammatory Cytokines Of Human Dental Pulp Cells Stimulated By LPS

Objective Human dental pulp cells(hDPCs)were stimulated with Escherichia coli(E.coli)lipopolysaccharide(LPS)to induce inflammation.To investigate the effects of Ginsenoside Rg1 on the inflammatory response and odontogenic differentiation of lipopolysaccharide-stimulated hDPCs.Methods1.hDPCs were cultured in media only or media containing different concentrations of ginsenoside Rg1(2.5,5,10 μmol/L)for 7 days.Cell proliferation was assessed by CCK-8 assay.2.hDPCs were treated with various doses of ginsenoside Rg1(2.5,5,10μmol/L)in the absence or presence of 1 μg/mL E.coli LPS for 1,3,5,7 days.Cell proliferation was assessed by CCK-8 assay.3.hDPCs were incubated with 5 μmol/L ginsenoside Rg1 in the absence or presence of 1 μg/mL E.coli LPS.After 7,14 days of osteogenic induction,alkaline phosphatase(ALP)activity and ALP staining were performed to evaluate the differentiation of hDPCs.After 21 days of osteogenic induction,the effects of ginsenoside Rg1 were evaluated by alizarin red stain.4.hDPCs were treated with LPS 3 hours before ginsenoside Rg1 treatment.TNF-α,IL-1β,IL-6m RNA levels were measured by real-time PCR.IL-6 protein levels were detected by ELISA.Results1.According to the results of the CCK-8 assay,the Optical Density(OD)value were increased in 5 μmol/L and 10 μmol/L Ginsenoside Rg1 group compared with control group on the fifth day.on the sixth day and the seventh day,the OD value were also increased in 2.5、5、10 μmol/L Ginsenoside Rg1 group compared with control group,the differences was statistically significant(p<0.0001).2.On fifth day,the OD value were increased in 2.5、5、10 μmol/L Ginsenoside Rg1+LPS group compared with the LPS group,the OD value were increased in 2.5、5、10 μmol/L Ginsenoside Rg1+LPS group compared with the LPS group on seventh day,and the differences was statistically significant(p<0.05).3.After osteogenic induction for 7 days,ALP activity in hDPCs of the Ginsenoside Rg1+LPS group and the LPS group showed a significant increase(p<0.05).After osteogenic induction for 7 and 14 days,ALP staining experiments showed that there was no significant difference between experimental groups.After osteogenic induction for 21 days,alizarin red stain experiment showed that there was more mineralized nodes in the Ginsenoside Rg1 groups compared with control groups.4.real-time PCR and ELISA showed that there was no significant difference between different experimental groups.ConclusionGinsenoside Rg1 can enhance the proliferation of hDPCs at an concentration of 5,10μmol/L with or without LPS.Ginsenoside Rg1 with a concentration of 5 μmol/L promotes the osteogenic differentiation of hDPCs.Ginsenoside Rg1 shows no anti-inflammatory effects on LPS stimulated hDPCs.