Objective To explore the effect of Ribes meyeri extracts on adipogenic and osteogenic differentiation of adipose-derived stem cells(ADSCs).Methods ADSCs of the p9 and p17 generations were used in the study,cell senescence was detected by β-galactosidase staining and expression of P16INK4a and P53 determined by real-time PCR.The cytotoxicity of Ribes meyeri on ADSCs was detected by CCK-8 assay,and the effect of Ribes meyeri on intracellular ROS content was detected by ROS kit.ADSCs of the p17 generation(senescent cells)treated with Ribes meyeri were subjected to adipogenic differentiation and ADSCs of the p9 generation(young cells)treated with Ribes meyeri were subjected to osteogenic differentiation.The expression of the adipogenic marker genes LPL and PPARγ,and the osteogenic marker genes OCN,RUNX2,and ALP were detected.Weighted gene co-expression network analysis(WGCNA)was constructed using undifferentiated,adipogenic and osteogenic differentiated and Ribes meyeri-treated adipogenic and osteogenic differentiated cells,respectively.KEGG enrichment of differentially expressed genes was performed for adipogenic differentiation of senescent cells and osteogenic differentiation of young cells treated with Ribes meyeri.Results The p17 generation of ADSCs was more senescent than p9 generations.The senescent ADSCs were more likely to differentiate into adipogenic cells,while young ADSCs were more likely to differentiate into osteoblasts.Ribes meyeri extract at concentration of 0.1 pg/μL promoted the cell proliferation of ADSCs,and Ribes meyeri at concentration of 1 ng/mL and 100 pg/mL decreased the level of ROS in ADSCs.Ribes meyeri inhibited the adipogenic differentiation of senescent ADSCs,and promoted osteogenic differentiation.The gene expression patterns of senescent and young ADSCs were different,and the gene expression pattern of young ADSCs was altered during the process of adipogenic differentiation.The effects of Ribes meyeri on adipogenic and osteogenic differentiation of ADSCs were correlated with TGF-β and Arg biosynthesis signaling pathways.Conclusion Ribes meyeri promotes the osteogenic differentiation of young ADSCs and inhibits the adipogenic differentiation of senescent ADSCs.
Ribes meyeriadipose mesenchymal stem cellsadipogenic and osteogenic differentiation
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