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本文旨在离体培养人肾小管上皮细胞株(human renal tubular epithelial cells,HKCs)上研究血管紧张素Ⅱ 1型受体阻断剂(angiotensin Ⅱ type 1 receptor blocker,ARB)坎地沙坦抗炎效应的机制。用流式细胞仪、Western blot、RT-PCR和ELISA技术分别检测HKCs的toll样受体4(toll-like receptor 4,TLR4)蛋白水平、血管紧张素II 1型受体(angiotensin Ⅱ type 1 receptor,AT1R)和磷酸化核因子-κB(nuclear factor-kappa B,NF-κB)p65蛋白、巨噬细胞趋化蛋白-1(macrophage chemoattractant protein-1,MCP-1)和RANTES的mRNA水平,以及MCP-1和RANTES在培养液中的蛋白浓度。结果显示,在离体培养的HKCs中,脂多糖(lipopolysaccharide,LPS)上调TLR4蛋白表达水平,增强NF-κB活化和下游炎症因子(包括MCP-1和RANTES)释放;坎地沙坦逆转LPS引起的TLR4表达的上调、NF-κB活化及MCP-1和RANTES的释放,但用siRNA下调AT1R表达并不改变坎地沙坦的这些效应。以上结果提示,坎地沙坦通过一条新的不依赖于AT1R的途径发挥抗炎效应。
This article aims to study the anti-inflammation of candesartan on human renal tubular epithelial cells (HKCs) in vitro by angiotensin Ⅱ type 1 receptor blocker (ARB) The mechanism of effect. The toll-like receptor 4 (TLR4) protein levels in HKCs were detected by flow cytometry, Western blot, RT-PCR and ELISA respectively. Angiotensin II type 1 receptor , AT1R) and p65 protein of nuclear factor-kappa B (NF-κB), macrophage chemoattractant protein-1 (MCP-1) and RANTES, Protein concentration of MCP-1 and RANTES in culture broth. The results showed that in cultured HKCs, lipopolysaccharide (LPS) up-regulated the expression of TLR4 protein, enhanced the activation of NF-κB and the release of downstream inflammatory cytokines including MCP-1 and RANTES. Candesartan reversed LPS-induced TLR4 expression, activation of NF-κB and release of MCP-1 and RANTES, but downregulation of AT1R expression with siRNA did not alter these effects of candesartan. The above results suggest that candesartan exerts anti-inflammatory effects through a new AT1R-independent pathway.