Effect of Wumen RMK-03TM active components on injury in LPS-treated nasal mucosa epithelial cells
吴门RMK-03TM喷剂有效组分对脂多糖引起的鼻黏膜上皮细胞损伤的作用研究

摘要

目的 研究吴门RMK-03TM喷剂有效组分对脂多糖（LPS）处理的鼻黏膜上皮细胞增殖和细胞炎性因子表达的影响。方法 体外培养人鼻黏膜上皮细胞HNEpC，用 0、1、2、4、8、16mg/L的RMK-03TM喷剂有效组分处理24h或48h，采用 CCK8法检测细胞增殖活性。将细胞分为空白对照组（不处理）、LPS 组（100 mg/L LPS 处理24h）和LPS + RMK-03TM喷剂有效组分组（100mg/L LPS 处理24h后加入4mg/L或8mg/L RMK-03TM喷剂有效组分作用24 h），采用CCK8法检测细胞增殖活性，ELISA法检测细胞中IL-1和TNF-α的表达，Western blot法检测细胞中蛋白p-IκBα、p-P65和血管紧张素转化酶2（ACE2）蛋白的表达。结果 RMK-03TM喷剂有效组分可促进鼻黏膜上皮细胞增殖，4-16mg/L 虫草素对鼻黏膜上皮细胞的增殖促进作用最明显。与空白对照组相比，LPS 组细胞增殖活性下降，IL-1β、TNF-α分泌含量和 p-IκBα、p-P65和ACE2蛋白的表达升高（P＜0.01）；与LPS 组相比，LPS + RMK-03TM喷剂有效组分组细胞增殖活性升高，IL-1β、TNF-α分泌含量和 p-IκBα、p-P65、ACE2蛋白的表达降低（P＜0.01）。结论 吴门RMK-03TM喷剂有效组分可以促进鼻黏膜上皮细胞增殖和抑制细胞炎症因子分泌，可能与抑制p-IκBα、p-P65和ACE2蛋白的表达有关。

Abstract

Objective To study the effect of Wumen RMK-03TM active components on the cell proliferation and expression of cytokines in LPS-induced nasal mucosa epithelial cells.
Methods HNEpC cells were cultured in vitro and the effect of Wumen RMK-03TM active components at different doses (0, 1, 2, 4, 8, 16mg/L) on the proliferation of human nasal mucosa epithelial cells was detected by CCK8 method. The cells were divided into four groups, blank control group (not treated), LPS group (treated with 100mg/L LPS for 24 hours), and LPS + RMK-03TM active components groups (4mg/L or 8mg/L components for 24hours after being treated with 100mg/L LPS for 24 hours). The proliferation of nasal mucosa epithelial cells was detected by CCK8 method. The contents of IL-1β and TNF-α a in the cell supernatant were detected by ELISA method. The expressions of p-IκBα, p-P65 and ACE2 proteins were detected by Western blot.
Results The proliferation activity of nasal mucosa epithelial cells increased after RMK-03TM active components treatment, especially the RMK-03TM active components at 416mg/L. Compared with the blank control group, the proliferation activity of LPS group was decreased, the contents of IL-1β, TNF-α and p-IκBα, p-P65, ACE2 protein were increased significantly (P＜0.05). Compared with LPS group, the proliferation activity of LPS + RMK-03TM active components groups was increased, the contents of IL-1β, TNF-α and p-IκBα, p-P65, ACE2 proteins were decreased (P＜0.05).
Conclusion   Wumen RMK-03TM active components can promote the proliferation of LPS induced nasal mucosa epithelial cells and inhibit the secretion of inflammatory cytokines. The possible mechanism is that RMK-03TM active components can inhibit the expression of the p-IκBα, p-P65 and ACE2 proteins.