不同浓度乙醇洗脱沙葱黄酮对脂多糖诱导的小鼠腹腔巨噬细胞的抗炎作用

摘要

This experiment was conducted to explore the anti-inflammatory effects of 35％ and 75％ ethanol elutions of Allium mongolicum Regel flavonoids (AMF) on lipopolysaccharide (LPS) -induced mouse peritoneal macrophage (MPM) .The effects of different concentrations (25, 50, 100, 200 and 400 μg/mL) of 35％and 75％ ethanol elutions of AMF on cell viability of MPM were examined by cell counting kit-8 assay.The blank control group, LPS stress model group, different concentration ethanol elutions of AMF groups (supplemented with 25, 50 and 100 μg/mL 35％ or 75％ ethanol elutions of AMF, respectively) were set up, and the effects different concentrations of ethanol elutions of AMF on cytokines and inflammatory factors of MPM were determined using Griess, real time PCR and enzyme-linked immune sorbent assay methods.The result showed that compared with the blank control group, supplemented with 25 to 100 μg/mL 35％ and 75％ ethanol elutions of AMF could significantly increase the cell viability of MPM (P<0.01) .Compared with the LPS stress model group, 35％ and 75％ ethanol elutions of AMF could significantly decrease the nitric oxide content and inducible nitric oxide synthase mRNA relative expression in MPM (P<0.01) , and showed a dose-dependent effect, and the effect of 75％ ethanol elutions of AMF was better than 35％ ethanol elutions of AMF;35％ethanol elutions of AMF could significantly decrease the contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in MPM (P<0.01) , and also could significantly decrease the TNF-α and IL-6 mRNA relative expression (P<0.01) ;75％ ethanol elutions of AMF could significantly decrease the contents of TNF-α in MPM (P<0.05 or P<0.01) , and also could significantly decrease the TNF-α and IL-6 mRNA relative expression (P<0.01) .In conclusion, 35％ and 75％ ethanol elutions of AMF can exert its anti-inflammatory effect via improving cell viability of MPM and affecting secretion of cytokines and inflammatory factors.%本试验旨在研究通过超声波法提取的35％和75％乙醇洗脱沙葱黄酮对脂多糖 (LPS) 诱导的小鼠腹腔巨噬细胞的抗炎作用.采用细胞计数试剂盒检测不同添加浓度 (25、50、100、200、400μg/mL) 的35％或75％乙醇洗脱沙葱黄酮对小鼠腹腔巨噬细胞活性的影响;设空白对照组、LPS应激模型组、不同浓度乙醇洗脱沙葱黄酮组 (分别加入25、50、100μg/mL 35％或75％乙醇洗脱沙葱黄酮溶液) , 采用Griess、实时荧光定量PCR及酶联免疫吸附测定法检测不同浓度乙醇洗脱沙葱黄酮对小鼠腹腔巨噬细胞细胞因子及炎性因子的影响.结果表明:与空白对照组相比, 35％、75％乙醇洗脱沙葱黄酮在25～100μg/mL添加浓度内可极显著提高小鼠腹腔巨噬细胞活性 (P<0.01) .与LPS应激模型组相比, 35％、75％乙醇洗脱沙葱黄酮均可极显著极显著降低小鼠腹腔巨噬细胞一氧化氮含量及诱导型一氧化氮合酶mRNA相对表达量 (P<0.01) , 并均呈剂量依赖效应, 且75％乙醇洗脱沙葱黄酮作用效果优于35％乙醇洗脱沙葱黄酮; 35％乙醇洗脱沙葱黄酮可极显著降低小鼠腹腔巨噬细胞肿瘤坏死因子-α (TNF-α) 、白细胞介素-6 (IL-6) 含量 (P<0.01) , 极显著降低TNF-α、IL-6 mRNA相对表达量 (P<0.01) ; 75％乙醇洗脱沙葱黄酮可显著或极显著降低小鼠腹腔巨噬细胞TNF-α含量 (P<0.05或P<0.01) , 极显著降低TNF-α、IL-6 mRNA相对表达量 (P<0.01) .由此可见, 35％、75％乙醇洗脱沙葱黄酮可通过提高小鼠腹腔巨噬细胞活性、调控细胞因子及炎性因子的分泌而发挥抗炎作用.