Interventional Effect Of Fucoidan On Inflammatory Response Of Human Bronchial Epithelial Cells

Objective:Fucoidan is a sulfated polysaccharide extracted from marine brown algae,which has been found to have various biological activities.This study aims to investigate the effects of fucoidan on the inflammatory response of bronchial epithelial cells,explore its related molecular pathways,and its comprehensive effects in respiratory inflammatory diseases.Methods:Experimental group and drug intervention: The experiment was divided into five groups: control group(without LPS stimulation),LPS group(10μg/m L),LPS +fucoidan low concentration group(50μg/m L),LPS + fucoidan medium concentration group(100μg/m L),LPS + fucoidan high concentration group(200μg/m L).The cells were stimulated with 10μg/m L LPS for 16 hours,and then treated with different concentrations of fucoidan for 8 hours according to the grouping.CCK8 method was used to observe the effect of fucoidan on LPS-induced cell damage,and the active oxygen detection kit was used to observe the effect of fucoidan and LPS on cell reactive oxygen species.Determine the inflammatory factors IL-6,IL-1β and TNF-α of each group by ELISA.QPCR was used to detect the transcription level of inflammatory factors,TLR4 and My D88 in each group.The expression of TLR4 and My D88 protein in cells of each group was detected by Western blot.The expression of TLR4 fluorescence in cells of each group was detected by immunofluorescence.Graphpad Prism 9.0 software was used for mapping and analysis,SPSS25.0software was used for one-way ANOVA.Image J was used for protein gray value analysis.Measurement data is expressed by(?)±s.P<0.05 indicates that the difference is statistically significant.Results:1.After being stimulated with 10μg/m L LPS for 16 hours,the viability of cells decreased and the expression of ROS increased.However,the concentration of fucoidan within the experimental range did not affect the viability of bronchial epithelial cells.After 8 hours of intervention with fucoidan in LPS stimulated cells,the cell viability significantly increased compared to the untreated group,and the expression of ROS decreased;2.In Elisa and q PCR experiments,compared with the control group,the inflammatory factors(IL-6,IL-1β,TNF-α)transcriptional level in LPS group increased significantly.After treatment with fucoidan,the transcription level decreased in a concentration dependent manner;3.Western blot experiments found that the expression of TLR4 and My D88 proteins in LPS group cells was significantly increased compared to the control group.After receiving fucoidan intervention,the expression levels of TLR4 and My D88 in cells were lower than those in LPS group,and the difference was statistically significant;4.In the cell immunofluorescence experiment,the expression of TLR4 fluorescence in LPS group cells was enhanced,significantly higher than that in the control group;The fluorescence intensity of TLR4 in cells treated with fucoidan was significantly lower than that in cells treated with LPS,and the difference was statistically significant.Conclusions:1.When LPS stimulates 16 HBE cells at an experimental concentration of10μg/m L for 16 hours,a cellular inflammation and oxidative stress model can be established;2.Fucoidan can improve cell damage by reducing the high levels of ROS produced by LPS stimulated cells,and may have antioxidant stress effects.Fucoidan may reduce the expression level of inflammatory factors induced by LPS by inhibiting the TLR4/My D88 pathway,thereby reducing the damage of inflammatory reactions to cells.3.Fucoidan can be considered for development as a marine derived drug used to curb respiratory inflammation.