摘要
为了探究柚皮素(NAN)对体外肠屏障炎性损伤和功能障碍的影响,本试验采用人肠上皮细胞系Caco-2和原代小鼠肠微血管内皮细胞(MIMVECs)共培养构建新型体外肠上皮屏障模型,使用1μg/mL脂多糖(LPS)诱导肠屏障炎性损伤,通过测定Caco-2细胞跨上皮电阻(TEER)和FITC-葡聚糖渗透量以评估柚皮素对肠屏障损伤的影响;通过分析Caco-2细胞炎性因子、紧密连接蛋白和NF-κB信号通路蛋白表达变化,进一步探究柚皮素保护肠屏障的机制。结果显示,柚皮素可显著抑制LPS诱导的共培养体系中上皮细胞单层TEER值的下降(P<0.05)和FITC-葡聚糖渗透量的升高(P<0.05);此外,柚皮素显著下调LPS诱导的共培养体系中上皮细胞炎性因子TNF-α(P<0.05)和IL-8(P<0.05)的分泌水平,上调紧密连接蛋白ZO-1(P<0.05)、Occludin(P<0.05)的表达,促进NF-κB信号通路蛋白IκB(P<0.05)的表达,抑制IκB(P<0.05)、p65(P<0.05)的磷酸化。结果表明,柚皮素可能通过抑制NF-κB信号通路的过度激活和炎性因子的分泌,增加紧密连接蛋白的表达,抑制LPS诱导的肠屏障炎性损伤和功能障碍。
To investigate the effect of naringenin(NAN)on inflammatory injury and dysfunction of intestinal barrier in vitro,this study co-cultured human intestinal epithelial cell line Caco-2 with primary mouse intestinal microvascular endothelial cells(MIMVECs)to establish a novel intestinal epithelial barrier model in vitro.Lipopolysaccharide(LPS,1μg/mL)was used to induce inflammatory injury of intestinal barrier.The effect of naringenin on intestinal barrier injury was evaluated by measuring the transepithelial electrical resistance(TEER)and FITC-dextran flux of Caco-2 cells.The mechanism of naringenin protecting intestinal barrier was further explored by analyzing expression levels of inflammatory factors,tight junction proteins and NF-κB signaling pathway proteins.The results showed that naringenin significantly inhibited the decrease of TEER(P<0.05)and the increase of FITC-dextran flux(P<0.05)in LPS-induced co-culture system.In addition,naringenin significantly down-regulated the secretion level of epithelial inflammatory factors TNF-α(P<0.05)and IL-8(P<0.05)in LPS-induced co-culture system,up-regulated the expression of tight junction proteins ZO-1(P<0.05)and Occludin(P<0.05),promoted the expression of NF-κB signaling pathway protein IκB(P<0.05),and inhibited the phosphorylation level of IκB(P<0.05)and p65(P<0.05).Thus,by inhibiting the overactivation of NF-κB signal pathway and the secretion of inflammatory factors,naringenin may increase the expression of tight junction proteins,and inhibit LPS-induced inflammatory damage and dysfunction of intestinal barrier.
作者
李欢欢
钟佳
周期律
卞艺斐
刘钟杰
LI Huan-huan;ZHONG Jia;ZHOU Qi-lv;BIAN Yi-fei;LIU Zhong-jie(College of Veterinary Medicine,China Agricultural University,Beijing 100193,China;Shandong Provincial Key Laboratory of Traditional Chinese Medicine for Basic Research,Shandong University of Traditional Chinese Medicine,Jinan 250355,China)
出处
《中国兽医杂志》
CAS
北大核心
2023年第1期130-135,共6页
Chinese Journal of Veterinary Medicine
基金
国家自然科学基金青年科学基金项目(31902315)。
