木犀草素·4,4′-联吡啶药物共晶对小鼠巨噬细胞RAW264.7的抗炎作用研究  

Anti-inflammatory Effect of the Luteolin·4,4′-dipyridy Co-crystal on Macrophage RAW264.7 in Mice

作　　者：刘立新[1] 邹冬玉 张羽男[1] 张强[2] 张楠楠[1] LIU Lixin;ZOU Dongyu;ZHANG Yunan;ZHANG Qiang;ZHANG Nannan(College of Pharmacy,Jiamusi University,Heilongjiang Jiamusi 154007,China;College of Public Health,Jiamusi University,Heilongjiang Jiamusi 154007,China)

机构地区：[1]佳木斯大学药学院,黑龙江佳木斯154007 [2]佳木斯大学公共卫生学院,黑龙江佳木斯154007

出　　处：《中国药房》2018年第5期602-606,共5页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81641120);黑龙江省卫生和计划生育委员会科研课题(No.2017-421)

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、80μmol/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40μmol/L浓度时细胞中诱导型一氧化氮合酶(i NOS)、环氧合酶2(COX-2)m RNA的表达,酶联免疫吸附法测定40μmol/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40μmol/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),i NOS和COX-2 m RNA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;40μmol/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i NOS和COX-2 m RNA表达水平及TNF-α、IL-6、NF-κB 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(10,20,40,80μmol/L),4,4′-dipyridy(10,20,40,80μmol/L)and luteolin·4,4′-dipyridy co-crystal(10,20,40,80μmol/L).The mRNA expression of iNOS and COX-2 in RAW264.7 cells at 40μmol/L were determined by qRT-PCR.The protein expression of TNF-αand IL-6 in RAW264.7 cells at 40μmol/L were determined by ELISA.The protein expression of NF-κB p65 in RAW264.7 cells at 40μmol/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 40μmol/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.

关 键 词：木犀草素 4 4′-联吡啶 药物共晶 小鼠巨噬细胞RAW264.7 抗炎作用 

分 类 号：R361.3[医药卫生—病理学]