| Objective:In this study,the serum of AECOPD,ACO patients were used to stimulate AM to establish an in vitro model of local inflammation,and PM2.5 was used to stimulate the inflammatory model in vitro.The expression of IL-6,IL-10 were detected,and the effects of PM2.5 stimulation on macrophages incubated with serum of AECOPD and ACO were investigated.Methods:Six groups were divided Control group,PM2.5 group,AECOPD group,ACO group,PM2.5+AECOPD group,PM2.5+ACO group.Enzyme linked immunosorbent assay(ELISA)was used to determine the concentrations of IL-6 and IL-10 in cell supernatant after cells were cultured for 6h.The concentrations of IL-6 and IL-10 and the expression of IL-6 and IL-10 were compared.Results:1.Comparison of clinical data: The expression of IL-6 in peripheral blood of ACO patients was higher than that of AECOPD patients(P<0.05).There was no significant difference between the two groups of patients in age,average smoking package years,FEV1%,white blood cell count,percentage of neutrophils,eosinophil percentage,IL-10 and intravenous hormone application(P>0.05).2.MH-S cell stimulated by PM2.5,the expression of IL-6 increased,and the difference was statistically significant compared with the control group(P<0.05).MH-S cells were stimulated by the serum of AECOPD patients and ACO patients,the expression of IL-6 increased,and the difference was statistically significant compared with the control group(P<0.05).The expression of IL-6 increased more obviously in the ACO group than that in the AECOPD group(P<0.05).Compared with the PM2.5 groups,the expression of IL-6 increased more obviously in MH-S cells simultaneously stimulated by PM2.5,AECOPD or ACO patients’ serum,The expression of IL-6 in PM2.5+ACO group increased more obviously than that in PM2.5+AECOPD group(P<0.05).3.PM2.5 stimulated MH-S cells,the expression of IL-10 decreased,and the difference was statistically significant compared with the control group(P<0.05).The serum of AECOPD patients and ACO patients stimulated MH-S cells,the expression of IL-10 decreased,and the difference was statistically significant compared with the control group(P<0.05).The expression of IL-10 decreased more obviously in the ACO group than that in the AECOPD group,but the difference was not statistically significant(P>0.05).Compared with the PM2.5 groups,the expression of IL-10 decreased more obviously in MH-S cells simultaneously stimulated by PM2.5,AECOPD or ACO patients’ serum(P<0.05).The expression of IL-10 in PM2.5+ACO group decreased more obviously than that in PM2.5+AECOPD group,but the difference wasn’t statistically significant(P>0.05).Conclusion:The expression of IL-6 increased and the expression of IL-10 decreased stimulated by PM2.5,The co stimulation of PM2.5 with the patient’s serum had stronger inflammatory effect,The inflammatory effect of PM2.5 on ACO patients may be higher than that of AECOPD patients.Objective:Silent information regulator 2-related enzymes 1(SIRT1)is thought to be involved in the regulation of cellular senescence and inflammation.Antagonistic crosstalk between SIRT1 and NF-κB signals present in many inflammatory diseases.In this study,PM2.5 was used to stimulate macrophages.SIRT1 activator SRT2104 and SIRT1 specific inhibitor EX527 were added,the protein expressions of SIRT1,NF-κB P65,P-P65,IκB,P-IκB and the expressions of IL-6,IL-10 were detected,the relationship between PM2.5 induced macrophage inflammation and SIRT1/NF-κB pathway were explored.Methods:The cell concentration were adjusted to 1×106/ m L,and the cells were cultured overnight in 6-well plates.After that,the cells were divided into Control group,PM2.5 group,PM2.5+SRT2104 group,PM2.5+ EX527 group,stimulated for 12 hours.Cells were collected and proteins were extracted.The expressions of SIRT1,NF-κB P-65,P-P65,IκB and P-IκB were detected by Western blot.The expressions of IL-6,IL-10 were detected by ELISA.Results:1.Compared with the control group,the expression of IκB and P-IκB decreased in PM2.5 group(P<0.05).The expressions of IκB and P-IκB in PM2.5+SRT2104group were decreased compared with the control group(P<0.05),but increased compared with the PM2.5 group(P<0.05),the expression of IκB and P-IκB in PM2.5+EX527group decreased compared with the control group and PM2.5group(P<0.05),no significant change compared with PM2.5 group.2.Compared with the control group,the expression of P65 and P-P65 increased in the PM2.5 group,and the change of P-P65 was statistically significant(P<0.05),the expression of P65 and P-P65 in PM2.5+SRT2104 group decreased compared with the control group and PM2.5 group,the changes of P-P65 was statistically significant(P<0.05).the expression of P65 and P-P65 in PM2.5+EX527 group increased compared with the control group and PM2.5 group,the difference of P-P65 expression was statistically significant(P<0.05).3.Compared with the control group,the expression of SIRT1 decreased in PM2.5group and PM2.5+EX527 group(P<0.05),but increased in the PM2.5+SRT2104group(P<0.05),compared with the PM2.5 group,the expression of SIRT1 increased in PM2.5+SRT2104 group but decreased in PM2.5+EX527group(P<0.05).4.The concentration of IL-6 and IL-10 in each group were detected.Compared with the control group,the concentration of IL-6 in PM2.5+SRT2104 group was decreased(P<0.05),compared with the PM2.5 group,the IL-6 concentration in the PM2.5+SRT2104 group was decreased(P<0.05),the concentration of IL-6 in PM2.5+EX527 group was not significantly changed compared with PM2.5 group.Compared with PM2.5+SRT2104 group,the concentration of IL-6 in PM2.5+EX527 group as increased(P<0.05).Compared with the control group,the concentration of IL-10 decreased in PM2.5 group,PM2.5+SRT2104 group and PM2.5+EX527 group(P<0.05).Compared with the PM2.5 group,the concentration of IL-10 increased in PM2.5+SRT2104 group,while decreased in PM2.5+EX527 group(P<0.05)compared with PM2.5+SRT2104 group,the expression of IL-10 in PM2.5+EX527 group decreased(P<0.05).Conclusion:PM2.5 can cause inflammation of MH-S cells through SIRT1/NF-κB pathway.SIRT1 may become a new target to inhibit PM2.5-induced inflammation of macrophages. |
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