大黄素对脂多糖诱导的角膜基质细胞表达细胞因子的影响  

Effects of Emodin on Expression of Cytokines Induced by Lipopolysaccharide in Corneal Fibroblasts

作　　者：陈国玲[1,2] 张晗[2] 刘艳丽[1] 孙洪义[2] 魏璐婉[1] 刘执玉[1] 

机构地区：[1]山东大学医学院解剖学教研室,济南250012 [2]山东大学眼科中心/第二医院眼科,济南250033

出　　处：《中国医学科学院学报》2009年第5期598-602,I0003,共6页Acta Academiae Medicinae Sinicae

基　　金：山东省教育厅科技计划项目(J08LH59)~~

摘　　要：目的观察大黄素对脂多糖(LPS)诱导的角膜基质细胞表达细胞因子的影响。方法原代培养角膜基质细胞,选取第4代细胞进行实验。根据在LPS刺激角膜基质细胞前是否先用40μmol/L大黄素培养细胞30 min,将细胞分为大黄素+LPS组和LPS组,分别在10μg/LLPS刺激角膜基质细胞前以及刺激后1、2、4、8 h收集细胞;或先用0、5、10、20和40μmol/L的大黄素培养细胞30 min后,再用10μg/L LPS刺激8 h。Western blot检测角膜基质细胞κB抑制因子-α(IκB-α)蛋白的表达;逆转录聚合酶链反应(RT-PCR)检测角膜基质细胞白细胞介素(IL)-6和IL-8的mR-NA表达。结果LPS刺激角膜基质细胞后1、2、4、8 h,细胞表达IκB-α蛋白均较刺激前明显下降(P<0.01);大黄素对LPS引起的角膜基质细胞IκB-α蛋白的降解有不同程度的抑制作用,且该作用呈一定的浓度依赖性(P<0.05)。LPS刺激引起角膜基质细胞IL-6和IL-8 mRNA表达升高,刺激后各时间点IL-6和IL-8 mRNA表达均较LPS刺激前明显升高,呈一定时间依赖性(P<0.01);大黄素对LPS诱导的角膜基质细胞IL-6和IL-8 mRNA表达有明显抑制作用,且该作用呈一定浓度依赖性(P<0.05)。结论大黄素能抑制体外培养的角膜基质细胞表达IL-6和IL-8,可能与其抑制IκB-α降解、促进NF-kB活化有关。Objective To investigate the effects of emodin on expression of cytokines induced by lipopolysaccharide （LPS） in cultured human corneal fibroblasts in vitro. Methods Primary human corneal fibroblasts of passages 4 were used in this research. Cells were treated with 10 μg/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 κB-α （IκB-α） 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）. Results Compared with cells without LPS treatment, IKB-α level significantly decreased in every time point after LPS challenge （P 〈 0.01 ）. Emodin inhibited the LPS-induced degeneration of IκB-α 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 ）. Conclusion 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 IKB-α and Suppressing the activation of nuclear factor-κB.

关 键 词：大黄素 脂多糖 细胞因子 角膜基质细胞 

分 类 号：R772.2[医药卫生—眼科]