PGC-1?–PPAR? Pathway Plays A Role In The Injury Mechanism Of PM2.5 Combined With Hypoxia-induced BEAS-2B Cells And The Protective Effect Of Yupingfeng Granules

Objective:Studies have found that Yupingfeng granules(YPF),a Chinese medicinal formula,have anti-inflammatory,antioxidant and immunomodulatory effects,but its mechanisms of action have not been well understood.In this study,we investigate the roles of PGC-1? and PPAR? in human bronchial epithelial cells(BEAS-2B)exposed to particulate matter with aerodynamic diameters <2.5 ?m(PM2.5)combined with hypoxia and the protective effect of YPF.Methods:1.To observe the influence of PM2.5,hypoxia and PM2.5 combined hypoxia on the expression of PGC-1? and PPAR? mRNA in BEAS-2B cells:cells were exposed to PM2.5 of 0,25,50,100?g/mL,hypoxia or PM2.5combined hypoxia respectively,and the expression of PGC-1? and PPAR?mRNA were detected with qPCR at 6,12,24 h after exposure to choose the time and concentration of PM2.5 combined hypoxia exposure.2.To investigate the protective mechanism of YPF on BEAS-2B cells exposed to PM2.5 combined hypoxia:after choosing the appropriate concentration of YPF with CCK8 method,setting up control group,PM2.5+hypoxia group,PM2.5+hypoxia+YPF group,PM2.5+ hypoxia+ siPGC-1?+YPF group,PM2.5+hypoxia+siCONTROL+YPF group.The level of mRNA and protein of PGC-1? and PPAR? were detected with qPCR and Western methods.The level of protein of ?-defension 2 and NF-?Bp65 were detected with Western methods.The level of protein of MCP-1 were detected with ELISA.Results:1.The levels of PGC-1? and PPAR? mRNA in different exposuregroups were lower significantly than that in the control group(P<0.01).Compared with the control group,the levels of PGC-1? and PPAR? mRNA and protein decreased in PM2.5+hypoxia group(P<0.01).Compared with PM2.5+hypoxia group,the levels of PGC-1?and PPAR? mRNA and protein decreased in PM2.5+hypoxia+YPF group(P<0.05).Compared with PM2.5+hypoxia+siCONTROL+YPF group,the levels of PGC-1? and PPAR? mRNA and protein decreased in PM2.5+hypoxia+siPGC-1?+YPF group(P<0.01).2.Compared with the control group,the levels of ?-defension 2 protein decreased in PM2.5+hypoxia group(P<0.01).Compared with PM2.5+hypoxia group,the levels of ?-defension 2 protein increased in PM2.5+hypoxia+YPF group(P<0.05).Compared with PM2.5+hypoxia+siCONTROL+YPF group,the levels of ?-defension 2 protein decreased in PM2.5+hypoxia+siPGC-1?+YPF group(P<0.01).3.Compared with the control group,the levels of MCP-1 protein increased in PM2.5+hypoxia group(P<0.01).Compared with PM2.5+hypoxia group,the levels of MCP-1 protein decreased in PM2.5+ hypoxia+YPF group(P<0.05).Compared with PM2.5+hypoxia+siCONTROL+YPF group,the levels of MCP-1 protein increased in PM2.5+hypoxia+siPGC-1?+YPF group(P<0.01).4.Compared with the control group,the levels of NF-?Bp65 protein increased in PM2.5+hypoxia group(P<0.01).Compared with PM2.5+hypoxia group,the levels of NF-?Bp65 protein decreased in PM2.5+ hypoxia+YPF group(P<0.05).Compared with PM2.5+hypoxia+siCONTROL+YPF group,the levels of NF-?Bp65 protein had no significant difference in PM2.5+hypoxia+siPGC-1?+YPF group.Conclusions: PM2.5 combined hypoxia can affect the expression of PGC-1? and PPAR? and induce an inflammatory reaction.YPF can improve these effects and may play a protective role through PGC-1?-PPAR? pathway.