【摘要】 为探讨细胞因子信号转导抑制因子3(suppressor of cytokine signaling 3, SOCS3)对高糖(high glucose, HG)诱导的大鼠肾小球系膜细胞RMC炎性因子表达的影响及机制，将RMC细胞分为6组：正常对照(normal control, NC)组、渗透压对照(osmotic control, OSM)组、 HG组、 HG+TAK-242(TLR4抑制剂)组、 HG+sh-SOCS3组和HG+sh-SOCS3+TAK-242组。CCK-8法检测细胞的存活率；流式细胞术检测细胞的凋亡情况；ELISA检测细胞培养上清液中炎性因子TNF-α、 IL-1β、 IL-6和COX2表达水平；qPCR法检测RMC细胞TLR4、 NF-κB p65和IκB-α mRNA的相对表达水平；Western blotting检测RMC细胞TLR4、 NF-κB p65、 p-p65、 IκB-α和p-IκB-α蛋白的表达量。结果显示，与NC组相比，HG组细胞存活率显著降低(P<0.01),细胞凋亡率，细胞上清液中TNF-α、 IL-1β、 IL-6和COX2表达水平，细胞中TLR4、p65、IκB-α mRNA相对表达水平和TLR4、 p-p65、 p-IκB-α蛋白表达水平均显著升高(均P<0.01);与HG组相比，HG+TAK-242组和HG+sh-SOCS3组细胞存活率显著升高(均P<0.05),细胞凋亡率，细胞上清液中TNF-α、 IL-1β、 IL-6和COX2表达水平，细胞中TLR4、p65、IκB-α mRNA表达水平和TLR4、 p-p65、 p-IκB-α蛋白表达水平均显著降低(均P<0.05); HG+sh-SOCS3+TAK-242组较HG+TAK-242组效果更加显著(均P<0.05)。由此，SOCS3可通过促进TLR4/NF-κB信号通路的激活上调HG诱导的大鼠系膜细胞炎性因子表达，加重炎症反应。更多还原

【Abstract】 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 IκB-α 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 expression of inflammatory factors in RMCs cultured with HG by activating the TLR4/NF-κB signaling pathway, thus aggravates the inflammatory responses.更多还原