[关键词]
[摘要]
本研究探究了紫云英苷对脂多糖(LPS)诱发的大鼠小肠上皮隐窝细胞(Rat intestinal epithelial cells,IEC-6)炎症模型抗炎作用。构建LPS诱发IEC-6细胞炎症模型,采用MTT法、酶联免疫吸附试验(ELISA)、实时荧光定量PCR和Western Blot法检测紫云英苷对IEC-6炎症细胞的存活率、相关炎症因子、基因及蛋白表达水平;结果显示,随着紫云英苷浓度的增加,IEC-6细胞的存活率逐渐增加,紫云英苷剂量(50 μg/mL、100 μg/mL)组存活率为90.68%和95.76%(p<0.05或p<0.01);酶联免疫吸附试验表明,与LPS组相比,紫云英苷50 μg/mL,100 μg/mL剂量显著抑制炎症因子,对IL-6炎症因子分泌水平抑制率分别为21.98%(p<0.05)和29.05%(p<0.01),mRNA表达水平分别降低了34.90%(p<0.05)和41.60%(p<0.01)。TNF-α炎症因子分泌水平抑制率分别为16.25%(p<0.05)和23.37%(p<0.01),mRNA表达水平分别降低了34.11%(p<0.05)和43.84%(p<0.01);蛋白通路方面,100 μg/mL的紫云英苷可显著抑制LPS诱导的IEC-6细胞NF-κB通路中P-IKKα/β降低了27.46%(p<0.05)、P-IκBα降低了41.52%(p<0.05)、P-p65降低了37.78%(p<0.01)。本探究为紫云英苷作为一种潜在缓解炎症性肠病药物的开发提供了可靠的依据。
[Key word]
[Abstract]
This study investigated the anti-inflammatory effect of Astragaloside on rat intestinal epithelial cells (IEC-6) model induced by lipopolysaccharide (LPS). A model of IEC-6 cell inflammation induced by LPS was established, and the survival rate, the related inflammatory factors, genes and protein expression levels of inflammatory IEC-6 cells were determined by MTT method, Enzyme-linked immunosorbent assay (ELISA), real-time fluorescence quantitative PCR and Western-blot, respectively. The results showed that the survival rate of IEC-6 cells increased gradually with the increase of Astragalin concentration. The survival rates of IEC-6 cells in different dose groups (50 μg/mL and 100 μg/mL) were 90.68% and 95.76% (p<0.05 and p<0.01), respectively. Enzyme-linked immunosorbent assay (ELISA) showed that compared with the LPS group, 50 μg/mL and 100 μg/mL of Astragalin significantly inhibited the secretion level of inflammatory factor IL-6, with inhibition rates of 21.98% (p<0.05) and 29.05% (p<0.01), respectively. The mRNA expression level of IL-6 was decreased by 34.90% (p<0.05) and 41.60% (p<0.01), respectively.The inhibitory rates of TNF-α secretion were 16.25% (p<0.05) and 23.37% (p<0.01), respectively, and the mRNA expression levels were decreased by 34.11% (p<0.05) and 43.84% (p<0.01), respectively. In terms of protein pathway, 100 g/mL of Astragalin showed significant inhibition of P-IKKα/β level 27.46% (p<0.05) and P-IκBα level 41.52% (p<0.05) in NF-κB pathway induced by LPS. This study provides a reliable basis for the development of Astragalin as a potential drug to relieve inflammatory bowel disease.
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[基金项目]
云南省科技厅重点项目(2018ZI001,2019ZG00905);云南省中青年学术和技术带头人后备人才项目(2018HB040)