Celastrol Ameliorates Inflammation in Human Retinal Pigment Epithelial Cells by Suppressing NF-κB Signaling
Celastrol Ameliorates Inflammation in Human Retinal Pigment Epithelial Cells by Suppressing NF-κB Signaling
复制标题
雷公藤红素通过抑制 NF-κB 信号传导改善人视网膜色素上皮细胞的炎症
DOI：
10.1089/jop.2018.0092
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发表时间：
2019
影响因子：
2.3
通讯作者：
Fu Shang
中科院分区：
文献类型：
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作者：
Jingyue Zhang;Kewen Zhou;Xinyu Zhang;Yeqi Zhou;Zhen Li;Fu Shang
目的：雷公藤红素是一种三萜奎宁类化合物，对多种疾病模型具有重要的生物学作用。在这项研究中，我们的目的是评估celastrol抑制脂多糖（LPS）诱导的视网膜色素上皮（RPE）细胞炎症的能力。方法：对人RPE （HRPE）细胞和ARPE-19细胞株进行单独或联合LPS处理。采用CCK-8法检测雷公藤红素对RPE细胞的细胞毒作用。通过流式细胞术和实时荧光定量PCR分别检测炎症细胞因子IL-6、IL-8和MCP-1的蛋白和mRNA水平。western blotting检测NF-κB信号通路磷酸化中间体（如i -κB α/β和p65）和MAPK信号通路磷酸化中间体（p38MAPK、SAPK/JNK和p42/p44MAPK）的水平。结果：在HRPE和ARPE-19细胞中，Celastrol显著抑制lps诱导的促炎细胞因子IL-6、IL-8和MCP-1的蛋白表达和mRNA表达水平。细胞活力和凋亡实验表明，在浓度小于1 μM时，celastrol对RPE细胞无明显的细胞毒作用，对RPE细胞凋亡有抑制作用。从机制上说，用celastrol预处理的RPE细胞表现出NF-κB通路调节因子、IKKα/β和i -κB α的磷酸化显著降低，随后P65失活，这表明celastrol通过抑制NF-κB信号通路来改善lps诱导的炎症。结论：雷公藤红素对RPE细胞具有较强的抗炎作用，在老年性黄斑变性的防治中具有潜在的应用价值。
Purpose:Celastrol is a triterpenoid quinine methide that exerts important biological effects on a variety of disease models. In this study, we aim to assess the ability of celastrol to inhibit lipopolysaccharide (LPS)-induced inflammation in retinal pigment epithelial (RPE) cells.Methods:Primary cultures of human RPE (HRPE) cells and ARPE-19 cell lines were treated with celastrol alone or in combination with LPS. The cytotoxic effect of celastrol on RPE cells was determined by the CCK-8 assay. Protein and mRNA levels of inflammatory cytokines, including IL-6, IL-8, and MCP-1, were detected by flow cytometry or by real-time fluorescent quantitative PCR, respectively. The levels of phosphorylated intermediates in the NF-κB signaling pathway (such as IκBα/β and p65) and MAPK signaling pathway (p38MAPK, SAPK/JNK, and p42/p44MAPK) were detected by western blotting.Results:Celastrol significantly inhibited LPS-induced expression of protein and mRNA expression levels encoding the proinflammatory cytokines, IL-6, IL-8, and MCP-1, in both HRPE and ARPE-19 cells. Cell viability and apoptosis assays revealed that celastrol had no apparent cytotoxic effect and it inhibited apoptosis of RPE cells at concentrations of less than 1 μM. Mechanistically, RPE cells that were pretreated with celastrol exhibited a substantial decrease in phosphorylation of the NF-κB pathway regulators, IKKα/β and IκBα, and subsequently inactivated P65, suggesting that celastrol ameliorates LPS-induced inflammation by suppressing the NF-κB signaling pathway.Conclusion:Our results provide evidence that celastrol is a potent anti-inflammatory agent in RPE cells and it may have potential applications in prevention and treatment of age-related macular degeneration.