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2022, Vol. 26 ›› Issue (13): 2000-2005Mechanism of paeoniflorin on bone marrow mesenchymal stem cells intervened by lipopolysaccharideLi Min, Yu Yang, Cheng Jiyan
Abstract: BACKGROUND: Bone marrow mesenchymal stem cells have strong potential application prospects in tissue engineering, cell therapy and gene therapy. More and more researches have focus on the effective improvement of survival rate and biological effect of bone marrow mesenchymal stem cell transplantation. OBJECTIVE: To investigate the effects of paeoniflorin on bone marrow mesenchymal stem cell proliferation, migration and the transforming growth factor-β1, ERK1/2 genes and proteins expression in bone marrow mesenchymal stem cells intervened by lipopolysaccharide. METHODS: The effect of paeoniflorin at different concentrations on the proliferation of bone marrow mesenchymal stem cells was detected by cell counting kit-8 assay. Scratch test was used to detect the effect of 10 μmol/L paeoniflorin on the migration ability of bone marrow mesenchymal stem cells. Real-time fluorescence quantitative PCR was used to detect the expression of collagen I, collagen III and Ki67 mRNA under the intervention of 10 μmol/L paeoniflorin. Bone marrow mesenchymal stem cells were divided into control group, lipopolysaccharide group (1 mg/L) and lipopolysaccharide (1 mg/L) + paeoniflorin (80 μmol/L) group. After 72 hours of intervention, real-time fluorescence quantitative PCR and western blot assay were used to detect the transforming growth factor-β1 and ERK1/2 (MAPK3/1) expression of genes and proteins in cells. Transforming growth factor-β1 protein location and semi-quantitative analysis in bone marrow mesenchymal stem cells were detected by immunofluorescence. RESULTS AND CONCLUSION: (1) Paeoniflorin increased the proliferation rate of bone marrow mesenchymal stem cells within a certain concentration (6.25-12.5 μmol/L). (2) The blank scratch area of the 10 μmol/L paeoniflorin group was significantly narrowed compared with the control group (P < 0.05). (3) Compared with the control group, the expression levels of collagen I, collagen III and Ki67 mRNA were significantly increased in the 10 μmol/L paeoniflorin group (P < 0.01). (4) Lipopolysaccharide significantly promoted transforming growth factor-β1, MAPK1, MAPK3 genes and transforming growth factor-β1, p-ERK1/2 protein expression (P < 0.01). After the addition of paeoniflorin, the up-regulated amount was significantly reduced (P < 0.01). (5) Based on the above experimental results, this study concludes that paeoniflorin can promote the proliferation, migration and the expression of collagen I, collagen III and Ki67 genes in bone marrow mesenchymal stem cells. Meanwhile, paeoniflorin can decline the expression of transforming growth factor-β1, ERK1/2 (MAPK3/1) genes and proteins in bone marrow mesenchymal stem cells intervened by lipopolysaccharide. Key words:stem cells, bone marrow mesenchymal stem cells, paeoniflorin, lipopolysaccharide, transforming growth factor-β1, extracellular regulated protein kinases |