CAJ | 학술논문

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30min,将细胞分为大黄素+LPS组和LPS组,分别在10μg/L LPS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40 μmol/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、8h,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.
목적 관찰대황소대지다당(LPS)유도적각막기질세포표체세포인자적영향.방법 원대배양각막기질세포,선취제4대세포진행실험.근거재LPS자격각막기질세포전시부선용40μmol/L대황소배양세포30min,장세포분위대황소+LPS조화LPS조,분별재10μg/L LPS자격각막기질세포전이급자격후1、2、4、8 h수집세포;혹선용0、5、10、20화40 μmol/L적대황소배양세포30 min후,재용10μg/L LPS자격8 h.Western blot검측각막기질세포κB억제인자-α(IκB-α)단백적표체;역전록취합매련반응(RT-PCR)검측각막기질세포백세포개소(IL)-6화IL-8적mR-NA표체.결과 LPS자격각막기질세포후1、2、4、8h,세포표체IκB-α단백균교자격전명현하강(P<0.01);대황소대LPS인기적각막기질세포IκB-α단백적강해유불동정도적억제작용,차해작용정일정적농도의뢰성(P<0.05).LPS자격인기각막기질세포IL-6화IL-8 mRNA표체승고,자격후각시간점IL-6화IL-8 mRNA표체균교LPS자격전명현승고,정일정시간의뢰성(P<0.01);대황소대LPS유도적각막기질세포IL-6화IL-8 mRNA표체유명현억제작용,차해작용정일정농도의뢰성(P<0.05).결론 대황소능억제체외배양적각막기질세포표체IL-6화IL-8,가능여기억제IκB-α강해、촉진NF-kB활화유관.
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, IκB-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 IκB-α: and suppressing the activation of nuclear factor-κB.