炎症介质对肾小管上皮细胞中PPARγ和辅调节因子表达的影响及机制.pdf

Recently, several studies have demonstrated that the expression of nuclear receptors (ie PPARs, RXR) and coregulators (ie SRC-1, SRC-2, SRC-3, PGC-1, NCoR) could be up-regulated in heart, liver, brain, adipose tissue by tumor necrosis factor-α (TNF-α), interlukin-1β (IL-1β) and other proinflammatory mediators. But the relationship between such proinflammatory mediators and PPARγ or its coregulators during inflammation of kidney is little known. We treated human renal tubular epithelial cells (HK-2) with TNF-α and IL-1β, and investigated the expression of NF-κB, PPARγ, coregulators and chemokine MCP-1 so as to study the role of PPARγ and coregulators in nephritis. In the first part of study, our results showed that TNF-α could up-regulate the expression of MCP-1 in dose- and time-dependent manners. The mRNA level of MCP-1 was significantly increased at 0.5h of 10ng/ml TNF-α stimulating （P0.05 ）, peaked at 4 hours （P0.01 ） and lasted to 8 hours （P0.05 ） . The excretion of MCP-1 protein in cell supernatant peaked at 24 hours, from 15.52± 1.05pg/ml to 40.5±0.97pg/ml （P0.01 ）. IL-1β (10ng/ml) could also up-regulate expression of MCP-1 protein from 19.33±2.33pg/ml to 160.56±2.8pg/ml （P0.01 ） at 8 hours ，then decreased to 50.82±1.25pg/ml at 24 hours （P0.05 ）. All these results indicated TNF-α and IL-1β could trigger inflammation response in HK-2 cells. In the second part of study, we found that both TNF-α and IL-1β could reduce the mRNA and protein levels of PPARγ in dose- and time-dependent manners. EMSA showed that the activities of PPARγ were down-regulated and NF-κB was up-regulated in time-dependent manner, and both of which peaked at 24 hours. These results demonstrated that TNF-α