Effects Of Chronic Obstructive Pulmonary Disease Patients’ Serum On Inflammation Of MH-S Cells Induced By PM2.5 And Mechanism Of Salubrinal

Part one: effects of serum from patients with chronic obstructive pulmonary disease on PM2.5-induced MH-S cell inflammationObjective: Establishment of model with MH-S cells.MH-S cells were stimulated by serum and PM2.5 in patients with chronic obstructive pulmonary disease(COPD).The aim of this study was to observe the changes of HDAC2 activity and IL-17 in MH-S cells.Methods: MH-S cells were stimulated with different concentrations of serum from patients with chronic obstructive pulmonary disease(COPD)and healthy people.The concentration of serum stimulation was determined to be 1% for 6 hours.Macrophages were stimulated with serum and PM2.5.They were divided into six groups: control group,PM2.5 group,COPD serum stimulate group,healthy volunteers serum stimulate group,PM2.5 and COPD serum costimulation group,PM2.5 and healthy volunteers serum costimulation group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of IL-17 in the supernatant of cells,and the activity colorimetric assay was used to detect the HDAC2 activity.The correlation between IL-17 and HDAC2 activity was analyzed by Pearson correlation analysis.Results: After stimulation of MH-S cells with PM2.5 and serum,the activity of HDAC2 was decreased and the secretion of IL-17 was increased compared with the control group.the change of HDAC2 activity decrease of MH-S cellsand IL-17 concentration increase in the serum stimulation group of patients with COPD was significant than that of the group of healthy volunteers serum stimulation group.and there was a significant difference between the two groups(P < 0.05).The activity of HDAC2 in MH-S cells decreased and the secretion of IL-17 increased more obviously in serum and PM2.5 costimulation groups,compared with PM2.5 group and serum alone group,the difference was statistically significant(P < 0.05).The IL-17 concentration were higher and the HDAC2 activity were lower in COPD patients’ serum and PM2.5 co-stimulation group compared with healthy volunteers’ serum and PM2.5 co-stimulation group(P < 0.05).There was a negative correlation between IL-17 and HDAC2(r = 0.786,P < 0.01).Conclusion: Simple PM2.5,serum stimulate could increase the secretion of IL-17 and decrease the activity of HDAC2 in MH-S cells.Serum can aggravate the effect of PM2.5 on MH-S cells.The effect of co-stimulation of COPD patients’ serum and PM2.5 on MH-S cells in was more obvious than that in healthy people serum and PM2.5 costimulation group.Part two: the role of Salubrinal in the inflammation of MH-S cells induced by PM2.5Objective: On the basis of the previous study.Salubrinal was used to interfere with MH-S,stimulated by PM2.5 to investigate the role of salubrinal in PM2.5 induced MH-S cell injury.Methods: The experiment was divided into 3 groups: control group:the blank control group was not given any stimulation at all.PM2.5 group: addingPM2.5 to the medium to make its final concentration 100 ug/ ml for 12 hours.PM2.5 and salubrinal group: salubrinal(30uMol salubrinal was added to the culture medium 1 hour ahead of time)and PM2.5,was stimulated for 12 hours.CHOP,GRP78 expression of MH-S was observed by immunofluorescence,IL-17 was measured by ELISA and activity Colorimetric determination of HDAC2.Results: After stimulation with PM2.5 for 12 hours,the CHOP fluorescence increased and enter into nucleus obviously,the GRP78 fluorescence increased and significantly in cytoplasm,the IL-17 content increased and the HDAC2 activity decreased compared with the blank control group,the difference was statistically significant(P<0.05).After the intervention of Salubrinal,the fluorescence of CHOP was lower than that of PM2.5,the fluorescence of GRP78 was higher than that of PM2.5,the content of IL-17 was decreased and the activity of HDAC2 was increased.The difference was statistically significant(P < 0.05).Conclusion: PM2.5 stimulation increased the expression of CHOP,GRP78,activated the ERS.Salibrinal inhibit the expression of CHOP in MH-S cells and enhanced the expression of GRP78.Salubrinal alleviated the damage effect of PM2.5 on MH-S Cell.