Chen Guo-Ling, Zhang Han, Liu Yan-Li, Sun Hong-Yi, Wei Lu-Wan, Liu Zhi-Yu
Department of Anatomy, School of Medicine, Shandong University, Ji'nan 250012, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2009 Oct;31(5):598-602.
To investigate the effects of emodin on expression of cytokines induced by lipopolysaccharide (LPS) in cultured human corneal fibroblasts in vitro.
Primary human corneal fibroblasts of passages 4 were used in this research. Cells were treated with 10 microg/L LPS for 1, 2, 4, or 8 hours, which were pretreated with or without emodin for 30 minutes before LPS challenge. The degeneration of inhibitor of kappaB-alpha (I kappaB-alpha) and the effect of emodin on it were analyzed by Western blot analysis with a specific antibody. The cellular abundance of the mRNA of interleukin (IL)-6 and IL-8 from corneal fibroblasts under different conditions was determined by reverse transcriptase polymerase chain reaction (RT-PCR).
Compared with cells without LPS treatment, I kappaB-alpha level significantly decreased in every time point after LPS challenge (P < 0.01). Emodin inhibited the LPS-induced degeneration of I kappaB-alpha by corneal fibroblasts in a dose-dependent manner (P < 0.05). Compared with cells without LPS treatment, the expressions of IL-6 and IL-8 mRNA significantly increased in every time point after LPS challenge (P < 0.01). At the same time, the expressions of the mRNA of IL-6 and IL-8 induced by LPS in corneal fibroblasts were also inhibited by emodin in a dose-dependent manner (P < 0.05).
Emodin can inhibit the expressions of IL-6 and IL-8 mRNA induced by LPS in corneal fibroblasts, which maybe via inhibiting the degeneration of I kappaB-alpha and suppressing the activation of nuclear factor-kappaB.
研究大黄素对体外培养的人角膜成纤维细胞中脂多糖（LPS）诱导的细胞因子表达的影响。
本研究采用第4代人角膜原代成纤维细胞。细胞在接受10μg/L LPS处理1、2、4或8小时前，先经或未经大黄素预处理30分钟。用特异性抗体通过蛋白质免疫印迹分析κB抑制蛋白α（IκB-α）的降解情况以及大黄素对其的影响。通过逆转录聚合酶链反应（RT-PCR）测定不同条件下角膜成纤维细胞中白细胞介素（IL）-6和IL-8 mRNA的细胞丰度。
与未用LPS处理的细胞相比，LPS刺激后各时间点IκB-α水平均显著降低（P<0.01）。大黄素以剂量依赖性方式抑制角膜成纤维细胞中LPS诱导的IκB-α降解（P<0.05）。与未用LPS处理的细胞相比，LPS刺激后各时间点IL-6和IL-8 mRNA表达均显著增加（P<0.01）。同时，大黄素也以剂量依赖性方式抑制角膜成纤维细胞中LPS诱导的IL-6和IL-8 mRNA表达（P<0.05）。
大黄素可抑制角膜成纤维细胞中LPS诱导的IL-6和IL-8 mRNA表达，其机制可能是通过抑制IκB-α降解并抑制核因子κB的激活。
