摘要：
目的:研究木犀草素·4,4'-联吡啶药物共晶的抗炎作用及机制.方法:以正常小鼠巨噬细胞RAW264.7为对照,以脂多糖(LPS)诱导的RAW264. 7细胞为炎症模型,采用MTT法检测不同浓度(10、20、40、80 μmol/L)的木犀草素、4,4'-联吡啶和木犀草素-4, 4'-联吡啶药物共晶作用细胞2 h后的细胞活性;荧光定量聚合酶链式反应法测定40 μmol/L浓度时细胞中诱导型一氧化氮合酶(iNOS)、环氧合酶2(COX-2)mRNA的表达,酶联免疫吸附法测定40 μmol/L浓度时细胞中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)蛋白表达,Western blot法测定40 μmol/L浓度时细胞中核转录因子p65 (NF-κB p65)蛋白表达.结果:与正常细胞比较, LPS诱导后RAW264. 7细胞活性明显降低(P<0. 01),iNOS和COX-2 mRNA表达水平及TNF-α、IL-6、NF-κB p65蛋白表达水平均明显升高(P<0. 01).木犀草素和木犀草素·4,4'-联吡啶药物共晶均能增强LPS诱导后RAW264. 7细胞活性(P<0. 05或P<0. 01), 且呈浓度依赖性;4,4'-联吡啶对LPS诱导后RAW264. 7细胞活性无明显影响;40 μmol/L浓度下,木犀草素和木犀草素·4, 4'-联吡啶药物共晶可使LPS诱导后细胞中iNOS和COX-2 mRNA表达水平及TNF-α、IL-6、NF-KB p65蛋白表达水平明显降低(P<0. 05或P<0. 01),其中木犀草素-4,4'-联吡啶药物共晶降低效果强于木犀草素(P<0. 05或P<0. 01).结论:木犀草素·4,4'-联吡啶药物共晶可能通过下调NF-κB信号来抑制炎症相关因子产生,其抗炎作用优于木犀草素.%OBJECTIVE: To study anti-inflammatory effect and mechanism of the luteolin · 4, 4' -dipyridy co-crystal.METHODS: Using macrophage RAW264. 7 of normal mice as control, the inflammation model was established with lipopolysaccharide (LPS) -induced RAW264. 7 cells. MTT assay was used to detect cells activity 2 h after treatment of different concentrations of luteolin (10, 20, 40, 80 μmol/L), 4, 4' -dipyridy (10,20, 40,80 μmol/L) and luteolin·4, 4' -dipyridy co-crystal (10, 20, 40, 80 μmol/L). The mRNA expression of iNOS and COX-2 in RAW264. 7 cells at 40 μmol/L were determined by qRT-PCR. The protein expression of TNF-α and IL-6 in RAW264. 7 cells at 40 μmol/L were determined by ELISA. The protein expression of NF-κB p65 in RAW264. 7 cells at 40 μmol/L were determined by Western bolt. RESULTS: Compared with normal cells, the activity of RAW264. 7 cells was decreased significantly after induced by LPS (P<0. 01); mRNA expression of iNOS and COX-2, protein expression of TNF-α, IL-6 and NF-κB p65 were increased significantly (P<0. 01). Both luteolin and luteolin · 4, 4' -dipyridy co-crystal could enhance the activity of RAW264. 7 cells after induced by LPS (P<0. 05 or P<0. 01) in concentration-dependent manner. 4, 4' -dipyridy had no significant effect on the activity of RAW264. 7 cells after induced by LPS. After luteolin and luteolin· 4, 4' -dipyridy co-crystal at 40 μmol/L, mRNA expression of iNOS and COX-2, protein expression of TNF-α, IL-6 and NF-κB p65 in RAW264. 7 cells after induced by LPS were decreased significantly (P<0. 05 or P<0. 01); the luteolin · 4, 4' -dipyridy co-crystal was better than luteolin (P<0. 05 or P<0. 01). CONCLUSIONS: The luteolin·4, 4' -dipyridy co-crystal can inhibit the generation of inflammatory factors by down-regulating NF-κB signal, and its anti-i nflammatory effect is better than luteolin.
