地塞米松对炎症环境下RAW264.7-L929共培养体系的影响  

Effect of Dexamethasone on RAW264.7-L929 Co-culture System in Inflammation

作　　者：陈远洋 陈晓珊 周全红[1] CHEN Yuanyang;CHEN Xiaoshan;ZHOU Quanhong(Department of ICU,Affiliated Shanghai Sixth People′s Hospital,Shanghai Jiao Tong University,Shanghai 200233,China)

机构地区：[1]上海交通大学附属第六人民医院,200233

出　　处：《医学研究杂志》2022年第11期30-34,共5页Journal of Medical Research

基　　金：国家自然科学基金资助项目(面上项目)(81771933)。

摘　　要：目的探讨在脂多糖(lipopolysaccharide,LPS)诱导的持续炎症环境下,地塞米松(dexamethasone,DEX)对RAW264.7-L929(小鼠巨噬细胞-成纤维细胞)共培养体系中细胞功能的影响。方法CCK-8法测试LPS与不同浓度的地塞米松对RAW264.7和L929细胞存活率的影响,选取对细胞存活率影响较小的药物浓度作为实验浓度;建立RAW264.7-L929共培养体系:分别以2×10^(5)个/孔与6×10^(5)个/孔的数量将两种细胞接种于Transwell小室和6孔板中。设立对照组、LPS组、DEX组、LPS+DEX组和DEX+LPS组,反转录PCR测试各组共培养体系中RAW264.7细胞炎性细胞因子IL-6、iNOS、IL-10 mRNA的表达。使用划痕实验评估24h后,各组共培养体系中L929细胞的迁移情况。结果筛选出×10^(-7) mol/L地塞米松及100ng/ml LPS作为实验处理浓度进行后续实验。地塞米松降低了持续炎症下共培养体系中RAW264.7细胞IL-6、iNOS mRNA的相对表达(P<0.05),但对IL-10 mRNA表达的影响差异无统计学意义。炎症环境下,地塞米松可以促进共培养体系中L929成纤维细胞的迁移(P<0.05),而对于地塞米松预先处理后再发生的炎症,地塞米松无促进成纤维细胞迁移的作用。结论地塞米松有助于共培养体系中炎症的缓解及成纤维细胞的迁移。Objective To investigate the effects of dexamethasone on cell function in the RAW264.7-L929(mouse macrophage fibroblast)co-culture system under the environment of continuous inflammation induced by lipopolysaccharide.Methods CCK-8method was used to test the effect of LPS and different concentrations of dexamethasone on RAW264.7 and L929 on cell viability.The drug concentration with little effect on cell viability was selected as the experimental concentration.In establishing RAW264.7-L929 co-culture system,two kinds of cells were co-cultured in Transwell chambers and six-well plates in the number of 2×10^(5)/well and 6×10^(5)/well,respectively.The control group,LPS group,DEX group,LPS+DEX group and DEX+LPS group were set up.The expression of inflammatory cytokines IL-6,iNOS and IL-10 mRNA of RAW264.7 in the co-culture system of each group was evaluated by RT-PCR.The migration rate of L929 cells in the co-culture system of each group after 24 hours was estimated by scratch test.Results ×10^(-7) mol/L dexamethasone and 100ng/ml LPS were selected as the experimental treatment concentration for subsequent experiments.Dexamethasone reduced the relative expression of IL-6 mRNA and iNOS mRNA(P<0.05)of RAW264.7 in co-culture system under persistent inflammation.But there was no significant difference in the relative expression of IL-10 mRNA.Dexamethasone could promote the migration(P<0.05)of L929 fibroblasts in the co-culture system under the inflammatory environment,but for the inflammation after dexamethasone pre-treatment,dexamethasone did not promote the migration of fibroblasts.Conclusion Dexamethasone alleviates inflammation and improves the migration of fibroblasts in the co-culture system under persistent inflammation.

关 键 词：地塞米松 炎症 巨噬细胞 成纤维细胞 共培养体系 

分 类 号：R34[医药卫生—基础医学]