槲皮素(Que)是一种在植物中发现的黄酮醇化合物,具有多种生物活性。坏死,一种特殊形式的程序性细胞死亡,在许多胃肠道疾病的发展中起着至关重要的作用。本研究旨在探讨Que能否通过调节凋亡信号通路减轻脱氧雪腐镰刀菌烯醇(DON)暴露后仔猪肠道损伤和屏障功能障碍。首先,对24头断奶仔猪进行了2×2因子设计,并分析了主要因素,包括Que(基础日粮或补充100mg/kgQue的日粮)和DON暴露(对照饲料或被4mg/kgDON污染的饲料)。饲喂21d后,仔猪被处死作为样本。接下来,在存在或不存在DON攻击(0.5μg/mL)的情况下,用或不用Que(10μmol/mL)预处理猪肠道上皮细胞系(IPEC-1)。饮食Que增加了体重,平均每日收益,和平均日采食量(p<0.05)。补牙提高了绒毛高度,DON暴露后断奶仔猪空肠中occludin和claudin-1蛋白表达较高(p<0.05),表明肠屏障功能增强(p<0.05)。饮食Que还下调了总受体相互作用蛋白激酶1(t-RIP1)的蛋白质丰度,磷酸化RIP1(p-RIP1),p-RIP3,总混合谱系激酶结构域样蛋白(t-MLKL),DON暴露后仔猪的p-MLKL(p<0.05)。此外,Que预处理增加了DON攻击后IPEC-1细胞上清液中的细胞活力并降低了乳酸脱氢酶(LDH)活性(p<0.05)。Que治疗还改善了上皮屏障功能,表现为更高的跨上皮电阻(TEER)(p<0.001),降低异硫氰酸荧光素标记的葡聚糖(FD4)通量(p<0.001),DON攻击后,occludin和claudin-1的分布更好(p<0.05)。此外,用Que预处理还抑制t-RIP1,p-RIP1,t-RIP3,p-RIP3,t-MLKL的蛋白质丰度,DON攻击后IPEC-1细胞中的p-MLKL(p<0.05)。总的来说,我们的数据表明,Que可以改善DON诱导的肠损伤和与抑制坏死凋亡信号通路相关的屏障功能障碍.
Quercetin (Que) is a flavonol compound found in plants, which has a variety of biological activities. Necroptosis, a special form of programmed cell death, plays a vital role in the development of many gastrointestinal diseases. This study aimed to explore whether Que could attenuate the intestinal injury and barrier dysfunction of piglets after deoxynivalenol (DON) exposure through modulating the necroptosis signaling pathway. Firstly, twenty-four weaned piglets were used in a 2 × 2 factorial design and the main factors, including Que (basal diet or diet supplemented with 100 mg/kg Que) and DON exposure (control feed or feed contaminated with 4 mg/kg DON). After feeding for 21 d, piglets were killed for samples. Next, the intestinal porcine epithelial cell line (IPEC-1) was pretreated with or without Que (10 μmol/mL) in the presence or absence of a DON challenge (0.5 μg/mL). Dietary Que increased the body weight, average daily gain, and average daily feed intake (p < 0.05) through the trial. Que supplementation improved the villus height, and enhanced the intestinal barrier function (p < 0.05) indicated by the higher protein expression of occludin and claudin-1 (p < 0.05) in the jejunum of the weaned piglets after DON exposure. Dietary Que also down-regulated the protein abundance of total receptor interacting protein kinase 1 (t-RIP1), phosphorylated RIP1 (p-RIP1), p-RIP3, total mixed lineage kinase domain-like protein (t-MLKL), and p-MLKL (p < 0.05) in piglets after DON exposure. Moreover, Que pretreatment increased the cell viability and decreased the lactate dehydrogenase (LDH) activity (p < 0.05) in the supernatant of IPEC-1 cells after DON challenge. Que treatment also improved the epithelial barrier function indicated by a higher transepithelial electrical resistance (TEER) (p < 0.001), lower fluorescein isothiocyanate-labeled dextran (FD4) flux (p < 0.001), and better distribution of occludin and claudin-1 (p < 0.05) after DON challenge. Additionally, pretreatment with Que also inhibited the protein abundance of t-RIP1, p-RIP1, t-RIP3, p-RIP3, t-MLKL, and p-MLKL (p < 0.05) in IPEC-1 cells after DON challenge. In general, our data suggest that Que can ameliorate DON-induced intestinal injury and barrier dysfunction associated with suppressing the necroptosis signaling pathway.