MT2A对脂多糖介导的人肺毛细血管内皮细胞损伤的保护作用

Protective Effect of MT2A on Lipopolysaccharide-mediated Injury of Human Pulmonary Capillary Endothelial Cells

目的:探索不同浓度的金属硫蛋2A(Metallothionein 2A, MT2A)对脂多糖(Lipopolysaccharid, LPS)介导的人肺微血管内皮细胞损伤的保护作用。方法:培养人肺毛细血管内皮细胞株(Human lung microvascular endothelial cells, HPMVECs),经过一定浓度LPS溶液进行刺激后,利用不同浓度的MT2A与对照组共同培养,一段时间后观察炎性介质IL-6、TNF-α释放的量及荧光显微镜观察组HPMVECs骨架形态变化。结果:各组TNF-α浓度均在0 h最低,随之逐渐升高,到6 h达到高峰;从各时间点来看,除0 h各组TNF-α浓度无显著差异外(F=0.717, P=0.549),其余各时间点B1、B2、B3均显著高于A组(均P<0.05)。各组中IL-6浓度均在0 h最低,A组在2 h达到高峰,随后逐渐下降;B1组在4 h达到高峰,随之下降;B2、B3组从0 h开始逐渐升高,到6 h达到峰值;从各时间点来看,除0 h各处理因素无显著差异外(F=2.341, P=0.092),其余各时间点B1、B2、B3均低于A组(均P<0.05)。A组6 h小时后纤维状肌动蛋白(F-actin)明显解聚,分布明显减少,应力纤维排列紊乱或者消失;B1组、B2组、B3组6 h小时后,与A组相比,F-actin的分布明显较多,应力纤维排列较为整齐。结论:LPS刺激人肺毛细血管内皮细胞有明显损伤效应,加入MT2A后细胞相关炎性因子释放量、细胞骨架损伤情况明显减轻,表明一定浓度的MT2A对LPS介导的肺毛细血管损伤有明显保护作用。

Objective: To explore the protective effects of different concentrations of metallothionein 2 A on human lung microvascular endothelial cell injury. Methods: Culture human lung capillary endothelial cell line, stimulated with a certain concentration of LPS solution, and then use different concentrations of MT2 A. Co-Cultivation with the control group, the amount of IL-6 and TNF-α released from the inflammatory medium and the morphology of the HPMVECs in the fluorescence microscope group were observed after a period of time. Results: The concentration of TNF-α in each group was the lowest at 0 h, then gradually increased, and reached a peak at 6 h.From the time points, except for 0 h, there was no significant difference in the concentration of TNF-α between the groups(F=0.717,P=0.549). At other time points, B1, B2, and B3 were significantly higher than those in group A(both P<0.05). The IL-6 concentration of each group was the lowest at 0 h, and the peak of group A peaked at 2 h, then gradually decreased. The peak of group B1 peaked at 4 h and then decreased. Groups B2 and B3 gradually increased from 0 h and peaked at 6 h. From each time point, there was no significant difference(F=2.341, P=0.092) except for 0 h treatment factor. B1, B2 and B3 were lower than group A at other time points(all P<0.05). After 6 hours in group A, fibrillating actin(F-actin) depolymerized, the distribution was significantly reduced, and the stress fibers were disordered or disappeared. After 6 hours in the B1, B2 and B3 groups, the distribution of F-actin was significantly more than that of the A group, and the stress fibers were arranged neatly. Conclusion: LPS stimulates human lung capillary endothelial cells to have obvious damage. After the addition of MT2 A, the release of cell-associated inflammatory factors and cytoskeletal damage were significantly reduced, indicating that certain concentrations of MT2 A have significant protective effects against LPS-mediated pulmonary capillary damage.