二十碳五烯酸和DHA差异抑制上皮间质转化和胶原沉积并缓解肠纤维化
EPA and DHA Differentially Inhibit Epithelial-to-Mesenchymal Transition and Decrease Collagen Deposition in Intestinal Fibrosis
冯政轩 1樊子怡 1陈诗薇 1谢政广 1陈凯阳 1周敏琪 1方剑1
作者信息
- 1. 绍兴文理学院医学院,浙江绍兴 312000
- 折叠
摘要
肠纤维化是炎症性肠病(inflammatory bowel disease,IBD)的严重并发症,然而目前尚未找到针对肠纤维化的治疗方案.ω-3 PUFAs 在其他器官中的抗纤维化作用已受到广泛关注,但在肠纤维化中的应用潜力仍有待发掘.本研究旨在探究ω-3 PUFAs 对肠纤维化的作用及机制.研究采用TNBS(2,4,6-trinitrobenzene sulfonic acid)诱导的肠纤维化小鼠模型,比较EPA、DHA 和鱼油对肠纤维化小鼠不同的抗纤维化作用.将40 只8 周龄雄性BALB/C 小鼠随机分为5 组.相较于DHA和鱼油组,EPA 干预能更有效缓解小鼠体重丢失,减少结肠长度缩短,降低疾病活动指数评分,并显著改善炎症反应(P＜0.05).ELISA 法检测发现,EPA 抑制促炎细胞因子TNF-α、IL-6 和IL-17 的表达,促进抗炎细胞因子IL-10 的表达.EPA 干预显著减轻了小鼠结肠的纤维化程度,下调了结肠中col1a2、co13a2 和羟脯氨酸的表达,并且结肠中EPA 含量与肠纤维化评分呈负相关(R2=0.7383,P＜0.0004;R2=0.4608,P＜0.0152).Western 印迹结果表明,EPA 上调p-AMPK、ULK1、LC3 和p62 的表达,下调mTOR 表达,同时抑制肠上皮细胞的上皮间质转化(epithelial-to-mesenchy-mal transition,EMT).qRT-PCR 分析显示,EPA 抑制α-SMA、TGF-β1、vimentin、TIMP-1(tissue inhibi-tors of metalloproteinases-1,TIMP-1)表达,增加MMP-9(matrix metalloproteinase-9,MMP-9)表达,从而抑制活化的肠间充质细胞,缓解细胞外基质(extracellular matrix,ECM)过度沉积.本研究发现,EPA 可恢复Th17/Treg 平衡,维持肠道免疫稳态,并抑制肠上皮细胞的EMT,调节TIMPs/MMPs 平衡,有效缓解肠纤维化小鼠结肠ECM 过度沉积.EPA 干预有望成为抗肠纤维化的新型辅助性防治策略,并提示自噬途径可能在抑制EMT 中发挥作用.
Abstract
Intestinal fibrosis is a serious complication of inflammatory bowel disease(IBD),however,no effective pharmacological treatment for intestinal fibrosis has been found yet.ω-3 PUFAs have re-ceived extensive attention for their anti-fibrotic effects in other organs,but their roles and mechanism in curing intestinal fibrosis are still unclear.In the present study,we compared the anti-fibrotic effects of EPA,DHA and fish oil in intestinal fibrosis mice using a TNBS(2,4,6-trinitrobenzene sulfonic acid)-induced intestinal fibrosis mouse model.Forty 8-week-old male BALB/C mice were randomly divided into five groups.Compared with the DHA and fish oil groups,EPA intervention more effectively alleviated weight loss,reduced colon length shortening,lowered disease activity index scores,and significantly im-proved inflammatory responses in the mice(P＜0.05).EPA inhibited the expression of pro-inflammatory cytokines TNF-α,IL-6,and IL-17,and promoted the expression of anti-inflammatory cytokine IL-10.EPA intervention significantly reduced the colonic histological severity of fibrosis,decreased col1a2,col3a2 and hydroxyproline expression in the colon,and EPA levels in the colon were negatively correlated with intestinal fibrosis scores(R2=0.7383,P＜0.0004;R2=0.4608,P＜0.0152).EPA up-regulated the expression of p-AMPK,ULK1,LC3 and p62,and also inhibited mTOR expression and the epithelial-to-mesenchymal transition(EMT)in intestinal epithelial cells.EPA inhibited α-SMA,TGF-β1,vimen-tin,TIMP-1(tissue inhibitors of metalloproteinases-1,TIMP-1)expression,and increased MMP-9(ma-trix metalloproteinase-9,MMP-9)expression,which inhibited activation of intestinal mesenchymal cells and alleviated excessive deposition of extracellular matrix(ECM).EPA restored Th17/Treg balance,maintained intestinal immune homeostasis,inhibited EMT in intestinal epithelial cells,regulated TIMPs/MMPs balance,and effectively alleviated the excessive deposition of colonic ECM in intestinal fibrosis mice.EPA is expected to be a novel adjuvant therapy for food-borne IBD with anti-fibrosis,and suggests that the autophagy pathway may play a role in inhibiting EMT.
关键词
炎症性肠病/Omega-3多不饱和脂肪酸/肠纤维化/自噬/上皮间质转化/胶原沉积Key words
inflammatory bowel disease(IBD)/Omega-3 polyunsaturated fatty acids(ω-3 PUFAs)/intestinal fibrosis/autophagy/epithelial-to-mesenchymal transition/collagen deposition引用本文复制引用
基金项目
浙江省基础公益研究计划(LGF18 H060006)
浙江省基础公益研究计划(LY21H260001)
浙江省医药卫生科技计划(2022KY1311)
浙江省医药卫生科技计划(2018KY819)
绍兴市市级医卫生类科技计划(2020A13064)
绍兴市市级医卫生类科技计划(2020A13064)
绍兴市市级医卫生类科技计划(2023SKY081)
国家级大学生创新创业训练计划(202210349030)
绍兴市市级科技计划(2023A14001)
绍兴文理学院科研启动基金(20210019)
出版年
2024
NETL
NSTL
维普
万方数据