SOCS3 promotes the high glucose-induced inflammatory factors expression in rat mesangial cells through TLR4/NF-κB signaling pathway
The aim of this study was to investigate the effect of suppressor of cytokine signaling 3(SOCS3)on the high glucose(HG)-induced inflammatory factors expression in rat glomerular mesangial cell(RMC)and the related mechanism.For this purpose,RMCs were divided into six groups:normal control(NC)group,osmotic control(OSM)group,HG group,HG+TAK-242(TLR4 inhibitor)group,HG+sh-SOCS3 group,and HG+sh-SOCS3+TAK-242 group.Cell viability was detected by CCK-8 assay and cell apoptosis was detected by FACS.The levels of inflammatory factors TNF-α,IL-1β,IL-6,and COX2 in cell culture supernatant were detected by ELISA.The mRNA expressions of TLR4,NF-κB p65,and IκB-α in RMC were detected by qPCR.The protein expressions of TLR4,NF-κB p65,p-p65,IκB-α,and p-IκB-α were detected by western blotting.The results showed that compared to those of NC group,the cell survival rate of the HG group was significantly decreased(P＜0.01),while the cell apoptosis rate,the levels of TNF-α,IL-1β,IL-6 and COX2 in cell culture supernatant,the mRNA expression of TLR4,p65 and IκBα and the protein expression of TLR4、p-p65 and p-IκB-α were significantly increased(all with P＜0.01).Compared to those of HG group,the cell survival rates of the HG+TAK-242 group and the HG+sh-SOCS3 group were significantly increased(both P＜0.05)while the apoptosis rate,the levels of TNF-α,IL-1β,IL-6 and COX2 in the cell culture supernatant,the mRNA expression of TLR4,p65,and IκB-α,and the protein expression of TLR4,p-p65,and p-IκB-α were significantly decreased(all with P＜0.05).The above changes were more significant in the HG+sh-SOCS3+TAK-242 group than the HG+TAK-242 group(all with P＜0.05).Taken together,SOCS3 up-regulates the expres-sion of inflammatory factors in RMCs cultured with HG by activating the TLR4/NF-κB signaling pathway,thus aggravates the inflammatory responses.
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