不同中药对中性粒细胞迁移微血管内皮细胞后溶菌酶释放量的影响

Effects of Different Chinese Herbs on the Lysozyme of Transendothelial Migration of Neutrophil

试验旨在探讨脂多糖(LPS)刺激大鼠肠黏膜微血管内皮细胞(RIMVECs)对跨内皮迁移中性粒细胞(PMNs)溶菌酶释放量的影响及白头翁、黄连、黄柏、秦皮、马齿苋的干预作用。选取0、0.1、1、10、100μg/mL LPS刺激RIMVECs,采用ELISA法检测12和24h细胞培养上清液中IL-6水平以确定后续试验所需LPS浓度。建立PMNs跨RIMVECs黏附模型和迁移的Transwell模型,采用细胞计数法、染色法、ELISA方法评价5种中药对LPS刺激RIMVECs后PMNs的黏附情况、迁移率及其溶菌酶释放量的影响。结果显示,与对照组相比,10μg/mL LPS作用RIMVECs后24h时的IL-6水平极显著升高(P<0.001);与LPS组相比,黄连、白头翁组的PMNs黏附率显著降低(P<0.05),溶菌酶释放量极显著升高(P<0.001),秦皮、白头翁组迁移率显著降低(P<0.05)。提示LPS刺激内皮细胞显著抑制了PMNs杀菌酶的释放,中药黄连、白头翁可以显著减轻这种抑制作用,PMNs杀菌酶的释放可能与内皮细胞的功能完整性有关,本试验为进一步筛选有效中药水溶性成分提供了理论依据。

To investigate the effect of lipopolysaccharide（LPS）stimulating rat intestinal mucous membrane microvascular cells（RIMVECs）on lysozyme content of transendothelial migration of neutrophil（PMNs）and the intervention of Pulsatillae（BTW）,Coptis chinensis（HL）,Cortex phellodendri（HB）,Cortex fraxini（QP）and Purslane（MCX）,0,0.1,1,10,100μg/mL LPS were selected to stimulate the RIMVECs in this experiment,and the IL-6 level in cells culture supernatants at 12 and 24 hwere determined by ELISA assay to determine the proper concentration of LPS for subsequent trials.To establish a transwell model of PMNs trans RIMVECs adherence and migration,the cell adhesion,migration rate and the release of lysozyme were evaluated by cell counts,staining and ELISA methods after LPS stimulating RIMVECs.The results showed that compared with the control group,the level of IL-6 at 24 hwas extremely significantly increased（P〈0.001）after RIMVECS was stimulated by 10μg/mL LPS.Compared with LPS group,the PMNs adhesion rate in HL and BTW groups,the migration rate of QP and BTW groups were sig-nificantly decreased（P〈0.05）,and the amount of lysozyme in HL and BTW groups were extremely significantly increased（P〈0.001）.The results showed that LPS stimulating endothelial cells could significantly inhibite the release of lysozyme bactericidal enzymes,intervention of Pulsatillae and Coptis chinensis could significantly reduce this inhibition.This study suggested that the release of lysozyme might be related to the functional integrity of endothelial cells,providing a theoretical basis for further screening of water-soluble components in effective Chinese herbal medicine.