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Hyperbaric oxygen combined with catechin inhibits the inflammatory response of human bone marrow stromal HS-5 cells by regulating TRAF6/NF-κB signaling pathway

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Author:
No author available
Journal Title:
Chinese Journal of Nautical Medicine and Hyperbaric Medicine
Issue:
5
DOI:
10.3760/cma.j.cn311847-20220122-00029
Key Word:
高压氧;儿茶素;TRAF6/NF-kB信号通路;人骨髓基质;HS-5细胞;炎症反应;Hyperbaric oxygen;Catechin;TLR6/NF-κB signaling pathway;Human bone marrow stroma;HS-5 cells;Inflammatory response

Abstract´╝Ü Objective:To discuss the effect of hyperbaric oxygen(HBO)combined with catechin(Can)on lipopolysaccharide(LPS)-induced inflammatory response of human bone marrow stromal HS-5 cells and its mechanism of action.Methods:A portion of normal HS-5 cells in good condition were assigned as control group(DMEM medium),and another portion of HS-5 cells were used to establish the bone marrow inflammatory cell model induced by LPS which were then divided into four groups for different interventions:the model group(1.0 μg/ml LPS was added into the medium),the HBO group(1.0 μg/ml LPS was added into the medium and then treated with 0.2 MPa HBO),the Can group(1.0 μg/ml LPS and 150 μmol/L Can were added into the medium)and the combined group(1.0 μg/ml LPS and 150 μmol/L Can were added into the medium and then treated with 0.2 MPa HBO). The proliferation of HS-5 cells in each group was detected by the CCK-8 method,and the expression levels of IL-1β,IL-6,TNF-α,and IL-10 in HS-5 inflammatory cell fluid were measured by ELISA. The apoptosis of HS-5 inflammatory cells in each group was detected by flow cytometry,and the expression levels of TLR6/NF-κB signaling pathway proteins in HS-5 inflammatory cells were detected by western blotting.Results:The results of the CCK-8 method showed that Can at the concentration of 150 μmol/L inhibited the proliferation of HS-5 cells induced by LPS at 1.0 μg/ml,which was used as a reference to establish an inflammatory cell model of bone marrow HS-5. The results of flow cytometry showed that the survival rate of HS-5 inflammatory cells in the combined group[(13.99±1.25)%]was significantly lower than that in the model group[(46.69±4.27)%],the HBO group[(39.28±3.74)%],and the Can group[(23.14±2.20)%],with statistically significant differences( P<0.05 or P<0.01). Compared with the model group,the HBO group,and the Can group,the expression levels of proteins(TLR6 and p-NF-κB)in HS-5 inflammatory cells in the combined group were significantly decreased( P<0.01). Conclusion:Can combine with HBO has a strong anti-inflammatory effect on HS-5 inflammatory cells,and its possible mechanism of action is to inhibit LPS-induced bone marrow inflammatory response by regulating the TLR6/NF-κB signaling pathway,which can provide new ideas for the treatment of bone injuries.

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