■非甾体抗炎药(NSAIDs)是一类解热处方药,镇痛药,抗炎,和抗血小板作用。然而,长期使用NSAIDs会破坏肠粘膜屏障,导致侵蚀,溃疡,出血,甚至穿孔。从柑橘的干燥果皮中提取纯的柑橘总黄酮(PTFC),显示对肠粘膜屏障的保护作用,机制不明确。
■在本研究中,我们使用双氯芬酸(7.5mgkg-1,i.g.)诱导NSAIDs相关肠道病变的大鼠模型。PTFC(50,75,100mg·kg-1d-1,i.g.)在初次双氯芬酸给药前9天,然后是过去5天的共同管理。通过蛋白质印迹和透射电子显微镜(TEM)鉴定外泌体,然后与IEC-6细胞共培养。长链非编码RNA(lncRNA)H19、自噬相关蛋白5(Atg5)、ZO-1,Occludin,通过定量实时PCR(qRT-PCR)检测Claudin-1。轻链3(LC3)-I的表达,LC3-II,通过蛋白质印迹测试ZO-1、Occludin和Claudin-1蛋白。通过免疫荧光技术检查外泌体和自噬体的定位。
■PTFC治疗减轻双氯芬酸诱导的NSAIDs大鼠肠黏膜机械屏障功能紊乱。与NSAIDs大鼠来源的外泌体共培养的IEC-6细胞具有最低水平的保护性自噬,和严重的肠屏障损伤。与从给予PTFC的大鼠中提取的外泌体共培养的细胞表现出自噬和肠粘膜机械屏障功能的改善。预防效果与施用的PTFC的浓度成比例。
■PTFC通过下调外泌体lncRNAH19和促进自噬改善NSAIDs诱导的肠粘膜损伤。
UNASSIGNED: Non-steroid anti-inflammatory drugs (
NSAIDs) are a class of prescription drugs with antipyretic, analgesic, anti-inflammatory, and antiplatelet effects. However, long-term use of
NSAIDs will disrupt the intestinal mucosal barrier, causing erosion, ulcers, bleeding, and even perforation. Pure total flavonoids from Citrus (PTFC) is extracted from the dried peel of Citrus, showing a protective effect on intestinal mucosal barrier with unclear mechanisms.
UNASSIGNED: In the present study, we used diclofenac (7.5 mg kg-1, i.g.) to induce a rat model of NSAIDs-related intestinal lesions. PTFC (50, 75, 100 mg·kg-1 d-1, i.g.) was administered 9 days before the initial diclofenac administration, followed by co-administration on the last 5 days. Exosomes were identified by western blotting and transmission electron microscopy (TEM), and then co-cultured with IEC-6 cells. The expression of long non-coding RNA (lncRNA) H19, autophagy-related 5 (Atg5), ZO-1, Occludin, and Claudin-1 were detected by quantitative real-time PCR (qRT-PCR). The expression of light chain 3 (LC3)-I, LC3-II, ZO-1, Occludin and Claudin-1 proteins was tested by western blotting. The localization of both exosomes and autophagosomes was examined by immunofluorescent technique.
UNASSIGNED: The treatment of PTFC attenuated intestinal mucosal mechanical barrier function disturbance in diclofenac-induced
NSAIDs rats. IEC-6 cells co-cultured with
NSAIDs rats-derived exosomes possessed the lowest levels of protective autophagy, and severe intestinal barrier injuries. Cells co-cultured with the exosomes extracted from rats administrated PTFC exhibited an improvement of autophagy and intestinal mucosal mechanical barrier function. The prevention effect was proportional to the concentration of PTFC administered.
UNASSIGNED: PTFC ameliorated
NSAIDs-induced intestinal mucosal injury by down-regulating exosomal lncRNA H19 and promoting autophagy.