Laboratory of Intestinal Adaptation and Recovery, Department of Paediatric Surgery, Dana-Dwek Children's Hospital, Tel Aviv Sourasky Medical Center, 6 Weizmann st, Tel Aviv 6423906, Israel.
Dept Pathology, Tel Aviv Sourasky Medical Center, Tel Aviv 6423906, Israel.
Nutrients. 2021 Mar 10;13(3):888. doi: 10.3390/nu13030888.
The aim of this study was to examine the anti-inflammatory and anti-apoptotic patterns of omega-3 polyunsaturated fatty acids (n-3 PUFAs) during methotrexate (MTX) induced intestinal damage in cell culture and in a rat model. Non-treated and treated with MTX HT 29 and HCT116cells were exposed to increasing doses of n-3 PUFAs and cell viability was evaluated using PrestoBlue assay. Male Sprague-Dawley rats were divided into 4 experimental groups: Control rats, CONTR+n-3 PUFA rats that were treated with oral n-3 PUFA, MTX rats were treated with MTX given IP, and MTX+n-3 PUFA rats were treated with oral n-3 PUFA before and following injection of MTX. Intestinal mucosal parameters and mucosal inflammation, enterocyte proliferation and apoptosis, TNF-α in mucosal tissue and plasma (ELISA), NF-κB, COX-2, TNF-α, Fas, FasL, Fadd, Bid, Bax and Bcl-2gene and protein levels were determined 72 h following MTX injection. Exposure of HT 29 and HCT116cells to n-3 PUFA attenuated inhibiting effects of MTX on cell viability. MTX-n-3 PUFA rats demonstrated a lower intestinal injury score and enhanced intestinal repair. A significant decrease in enterocyte apoptosis in MTX+n-3 PUFA rats was accompanied by decreased TNF-α, FAS, FasL, FADD and BID mRNA levels. Decreased NF-κB, COX-2 and TNF-α levels in mucosa was accompanied by a decreased number of IELs and macrophages. n-3 PUFAs inhibit NF-κB/COX-2 induced production of pro-inflammatory cytokines and inhibit cell apoptosis mainly by extrinsic pathway in rats with MTX-induced intestinal damage.
本研究旨在探讨ω-3 多不饱和脂肪酸（n-3 PUFAs）在细胞培养和大鼠模型中对甲氨蝶呤（MTX）诱导的肠道损伤的抗炎和抗凋亡作用模式。非处理和 MTX 处理的 HT 29 和 HCT116 细胞暴露于递增剂量的 n-3 PUFAs 后，使用 PrestoBlue 检测法评估细胞活力。雄性 Sprague-Dawley 大鼠分为 4 个实验组：对照组大鼠、口服 n-3 PUFA 处理的 CONTR+n-3 PUFA 大鼠、腹腔注射 MTX 的 MTX 大鼠和腹腔注射 MTX 前和后口服 n-3 PUFA 的 MTX+n-3 PUFA 大鼠。在 MTX 注射后 72 小时，测定肠黏膜参数和黏膜炎症、肠上皮细胞增殖和凋亡、黏膜组织和血浆中的 TNF-α（ELISA）、NF-κB、COX-2、TNF-α、Fas、FasL、Fadd、Bid、Bax 和 Bcl-2 基因和蛋白水平。HT 29 和 HCT116 细胞暴露于 n-3 PUFA 可减轻 MTX 对细胞活力的抑制作用。MTX+n-3 PUFA 大鼠的肠道损伤评分较低，肠道修复能力增强。MTX+n-3 PUFA 大鼠的肠上皮细胞凋亡明显减少，同时 TNF-α、FAS、FasL、FADD 和 BIDmRNA 水平降低。黏膜中 NF-κB、COX-2 和 TNF-α 水平降低伴随着 IELs 和巨噬细胞数量减少。n-3 PUFAs 可抑制 NF-κB/COX-2 诱导的促炎细胞因子产生，并通过外源性途径抑制 MTX 诱导的肠道损伤大鼠的细胞凋亡。
