五灵胶囊中部分化学成分对脂多糖诱导枯否细胞释放炎症因子的影响

Influence of Part Chemical Components in Wuling Capsules on the Expression of ERK and NF-КB Signal Pathway in Kupffer Cells Induced by Lipopolysaccharide

关键词:  五灵胶囊  有效成分  枯否细胞  炎性因子  核因子-κB

Key words:  Wuling capsules  active ingredient  Kupffer cell  inflammatory factor  nuclear factor-kappa B

摘要:

目的 观察五灵胶囊(Wuling capsules，WL)中部分化学成分对脂多糖(lipopolysaccharider，LPS)诱导大鼠原代枯否细胞(Kupffer cell，KC)表达ERK(extracellular signal-regulated kinase)和核因子(nuclear factor-kappa B，NF-κB)调节炎症因子和介质的作用。方法 分离大鼠KC，采用60 μg·L^-1 LPS诱导KC分泌炎症因子及NOS，ELISA测定TNF-α、IL-6、IL-8，比色法测定NOS，Western blot检测全蛋白ERK、p-ERK、NF-κBp50、NF-κBp65、p-NF-κBp65、p-IκK、p-IκB及核、浆蛋白NF-κBp65、p-NF-κBp65的变化。结果 WL明显下调LPS活化KC表达p-ERK、p-NF-κBp65、p-IκK、p-IκB蛋白，模拟WL混合成分(Mix)及6单体成分明显下调LPS活化KC表达p-ERK和p-NF-κBp65信号通路蛋白。各受试药物组均能降低核内p-NF-κBp65表达，减少p-NF-κBp65入核抑制炎性基因转录，降低活化KC过量分泌TNF-α、IL-6、IL-8和NOS。结论 WL中五味子醇甲、五味子乙素、隐丹参酮、丹参酮ⅡA、柴胡皂苷D是干预LPS诱导KC表达ERK、NF-κB信号通路蛋白，抑制炎性因子与介质基因转录而减少TNF-α、IL-6、IL-8和NOS分泌的有效成分。

Abstract:

OBJECTIVE The effects of Wuling capsules and their part of chemical components on lipopolyccharider(LPS) inducing the expression of extracellular signal-regulated kinase(ERK) and nuclear factor-kappaB (NF-κB) signaling pathway to regulate inflammatory factor and medium in rat primary Kupffer cells (KC) were observed. METHODS Liver cells and KC were separated, the promoting inflammatory factor and nitrous oxide (NO) were secreted in KC induced by 60 μg·mL^-1 LPS, and the TNF-α, IL-6, IL-8 were determined by ELISA, and the nitrous oxide producing quantity were detected by colorimetry, the expression change of NF-κBp50, NF-κBp65, p-NF-κBp65, IκB, p-IκB, TNF-α albumen in KC were examined by western blot. RESULTS Wuling capsules could obviously down-regulate the expression of NF-κB signal pathway albumen in KC activated by LPS, and the mix ingredients of imitation Wuling casules and six monomer components produced differently to regulate effects on the expression level of p-ERK and p-NF-κBp65 signal molecule protein. Cytoplasm and nuclear distribution test showed the expression level of p-ERK and p-NF-κBp65 in nuclear of KC activated by LPS was significantly lowered, the p-NF-κBp65 import nuclear was significantly attenuated and evoked gene transcription was inhibited, then the contents of secreting TNF-α, IL-6, IL-8, NO by activation KC were significantly lowered at the same time. CONCLUSION The schizandrol A, schizandrin B, cryptanshinono, tanshinone ⅡA, saikosaponins D in Wuling capsules are the active component of Wuling capsules in treatments of inflammation response of chronic hepatitis.