目的: 探索体外模拟糖尿病状态下外源性IL-33对心肌成纤维细胞的作用。方法: 体外分离培养新生乳鼠心脏成纤维细胞，设立对照组（DMEM/F12+30 mmol/L甘露醇），高糖组（DMEM/F12+30 mmol/L葡萄糖），高糖+高迁移率族蛋白1（HMGB1）组（1 μg/mL），高糖+HMGB1+IL-33组（3 ng/mL）。经不同处理后，蛋白质印迹检测胶原蛋白Ⅰ、IL-33、二酰基甘油激酶ζ（DGKζ）表达量的变化；ELISA检测二酰基甘油（DAG）的分泌释放，以及蛋白激酶Cβ（PKCβ）活性的改变。结果： 心肌成纤维细胞经高糖处理后胶原蛋白Ⅰ生成增加，IL-33产量降低，HMGB1使上述情况进一步加剧，心肌细胞内DGKζ的表达下调，引起DAG的升高，并进一步促进PKCβ的激活，引发心肌纤维化。加入外源性IL-33对上述过程具有抑制作用，可抗心肌纤维化。结论： 外源性IL-33在心肌成纤维细胞模拟糖尿病状态下起抗纤维化作用。
Abstract
Objective： To investigate the role of IL-33 on cardiac fibrosis under high glucose treatment on cardiac fibroblasts. Methods：Isolated rat cardiac fibroblasts were divided into following groups: control (DMEM/F12+30 mmol/L mannitol), high glucose(DMEM/F12+30 mmol/L glucose),high glucose+HMGB1(1 μg/mL), high glucose +HMGB1+IL-33 (3 ng/mL).The expression of the collagen I, IL-33 and DGK ζ in the cardiac fibroblasts were evaluated by western blotting. The DAG and the activity of PKCβ was detected by ELISA. Results：When cardiac fibroblasts challenged with high glucose, the collagen I generation was increased and the production of IL-33 was decreased. The effect of high glucose was exaggerated by HMGB1. In addition, the DGKζ expression was decreased, DAG content was increased and further promoted the activation of PKCβ.Exogenous IL-33 attenuated above effects. Conclusion：Exogenous IL-33 plays a key role in anti-cardiac fibrosis induced by high glucose.
关键词
IL-33 /
糖尿病心肌病 /
肌成纤维细胞 /
胶原蛋白 /
纤维化
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