Effect of 1-Deoxynojirimycin(DNJ)from mulberry extract on MC3T3-E1 cells under high glucose environment
Objective To investigate the effect of 1-Deoxynojirimycin(DNJ)on the proliferation and osteo-genic differentiation of MC3T3-E1 cells in high glucose environment.Methods The P4 generation MC3T3-E1 cells were taken,and the effects of different concentrations of DNJ(10 mmol/L,1 mmol/L,100 μmol/L,10 μmol/L,1 μmol/L,100 nmol/L,10 nmol/L and 1 nmol/L)on the proliferation of osteo-genic precursor cells were detected by CellDounting Kit-8(CCK-8)in a high glucose environment of 50 mmol/L,and the concentration of DNJ in subsequent experimental studies was screened.MC3T3-E1 cells cultured to logarithmic growth phase were selected and divided into blank group(complete medium),high glucose group(complete medium with 50 mmol/L glucose concentration)and experimental group[complete medium with 50 mmol/L glucose concentration+different concentrations of DNJ(100 μmol/L,10 μmol/L,1 μmol/L)solution].Flow cytometry was used to detect cell apoptosis and reactive oxygen species production.The contents of AGEs,IL-1β,IL-6 and TNF-α were detected by ELISA kit.The early osteogenic ability of MC3T3-E1 cells were detected by alkaline phosphatase staining and activity.The late osteogenic ability of MC3T3-E1 cells were detected by alizarin red staining and quantitative detection.The mRNA expressions of IL-1β,IL-6,TNF-α,Bax,Bcl-2,IGF-1,ALP,OCN,OSX,Col-1 and Runx2 were detected by RT-PCR.Results In the high glucose environment,100 μmol/L,10 μmol/L and 1 μmol/L DNJ can significantly promote the proliferation of MC3T3-E1 cells,so it is used for subsequent research.Compared with the high glucose group,the apoptosis rate,reactive oxygen species and the contents of AGEs,IL-1β,IL-6 and TNF-α in the cell supernatant of the experimental group weredecreased,and the difference was statistically significant(P＜0.05).The results of RT-PCR showed that compared with the high glucose group,the mRNA expression of Bax,IL-1β,IL-6 and TNF-α in the experimental group was significantly decreased,and the mRNA expression of Bcl-2,IGF-1,ALP,OCN,OSX,Col-1 and Runx2 was significantly increased,the difference was statistically significant(P＜0.05).Conclusion In the high glucose environment,DNJ can inhibit the apoptosis of mouse pre-osteoblast MC3T3-E1 and the production of in-flammatory factors within a certain concentration range,and promote the differentiation of osteoblasts.
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