摘要
本试验以脂多糖(LPS)诱导的小鼠巨噬细胞RAW264.7为试验材料,研究水蔓菁总黄酮的体外抗炎活性及其机制.采用MTT法检测水蔓菁总黄酮的细胞毒性,中性红试验测定细胞吞噬能力,Griess法检测一氧化氮(NO)的生成量,酶联免疫吸附测定(ELISA)法检测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、前列腺素E2(PGE2)、诱导型一氧化氮合酶(iNOS)和环氧合酶-2(COX-2)的生成量,Western blot法检测核因子-κB(NF-κB)信号通路相关蛋白表达量.结果显示:质量浓度为200 μg/mL的水蔓菁总黄酮对小鼠巨噬细胞 RAW264.7 无显著毒性(P＞0.05).不同质量浓度(25、50、100、200、300、400 μg/mL)的水蔓菁总黄酮均能显著或极显著增强LPS诱导的小鼠巨噬细胞RAW264.7的吞噬能力(P＜0.05或P＜0.01),显著或极显著抑制NO、IL-1、IL-6、TNF-α、PGE2、iNOS和COX-2的生成量及磷酸化核因子-κB p65(p-NF-κB p65)和磷酸化核因子-κB抑制蛋白α(p-IκBα)的蛋白表达量(P＜0.05或P＜0.01),并能显著或极显著增强核因子-κB抑制蛋白α(IκBα)的蛋白表达量(P＜0.05或P＜0.01).综上可知,水蔓菁总黄酮能增强小鼠巨噬细胞的吞噬能力,并能够通过抑制细胞内iNOS、COX-2、NO和炎症因子的生成和调控NF-κB信号通路相关蛋白表达来发挥体外抗炎活性.
Abstract
The purpose of this study was to investigate the in vitro anti-inflammatory activity and mechanism of total flavonoids from Veronica linariifolia by lipopolysaccharide(LPS)-induced mouse macrophages RAW264.7.MTT assay was used to detect the cytotoxicity of total flavonoids from Veronica linariifolia;the phagocytosis capacity was determined by neutral red assay;the production of nitric oxide(NO)was detected by Griess method;enzyme-linked immunosorbent assay(ELISA)method was used to detect the production of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),prostaglandin E2(PGE2),inducible nitric oxide synthase(iNOS)and cyclooxygenase-2(COX-2);the expression levels of nuclear fac-tor-κB(NF-κB)signaling pathway-associated proteins were assayed by Western blotting method.The results showed that the total flavonoids from Veronica linariifolia in the mass concentration of 200 μg/mL had no sig-nificant cytotoxicity to mouse macrophages RAW264.7(P＞0.05).Different mass concentrations(25,50,100,200,300 and 400 μg/mL)of total flavonoids from Veronica linariifolia could significantly or extremely significantly enhance the phagocytosis capacity of mouse macrophages RAW264.7 induced by LPS(P＜0.05 or P＜0.01),significantly or extremely significantly inhibit the production of NO,IL-1β,IL-6,TNF-α,PGE2,iNOS,COX-2 and the protein expression levels of phosphorylated nuclear factor-KB p65(p-NF-KB p65)and phosphorylated inhibitor of nuclear factor-κBα(p-IκBα)(P＜0.05 or P＜0.01),and significantly or extremely significantly enhance the protein expression level of inhibitor α of nuclear factor-κB(IκBα).In conclusion,the total flavonoids from Veronica linariifolia can enhance the phagocytosis function of mouse macrophages and exert anti-inflammatory activity in vitro by inhibiting the production of iNOS,COX-2,NO and inflammatory factors,and regulating the expression of NF-κB signaling pathway-associated proteins.[Chinese Journal of Animal Nutrition,2024,36(6):3963-3972]
基金项目
山西省2020年度职业院校教师素质提高计划中高职教师素质协同提升项目(CXTD2020-11)
山西药科职业学院科研资助项目(2020106)
山西药科职业学院教师发展基金专项资助项目()
维普
万方数据