1 Department of Pediatrics and Infantile Neuropsychiatry, Pediatric Gastroenterology and Liver Unit, Sapienza University of Rome, Viale Regina Elena 324, 00161 Rome, Italy.
2 Division of Health Protection Technologies, Territorial and Production Systems Sustainability Department, ENEA, Via Angullarese 301, 00123 Rome, Italy.
Benef Microbes. 2018 Apr 25;9(3):389-399. doi: 10.3920/BM2017.0078. Epub 2018 Apr 10.
Current research into original therapies to treat intestinal inflammation is focusing on no-drug therapies. KLD is a mixture of krill oil (KO), probiotic Lactobacillus reuteri (LR), and vitamin D (VitD3). The aim of this study was to assess in vitro and in vivo the potential cooperative effects of KLD in reducing gut inflammation. Colorectal adenocarcinoma cell lines, CACO2 and HT29, and C57BL/6 mice were used for in vitro and in vivo analyses, respectively. Cells were exposed to cytomix (interferon gamma + tumour necrosis factor alpha (TNF-α)) to induce inflammation or co-exposed to cytomix and KO, LR and VitD3 alone or to cytomix and KLD. Animals were treated for 7 days with dextran sodium sulphate (DSS) to induce colitis or with DSS and KLD. In vitro assays: F-actin expression was analysed by immunofluorescence; scratch test and trans-epithelial electric resistance test were performed to measure wound healing; adhesion/invasion assays of adhesive and invasive Escherichia coli (AIEC) bacteria were made; mRNA expression of TNF-α, interleukin (IL)-8 and vitamin D receptor (VDR) was detected by quantitative PCR. In vivo assays: body weight, clinical score, histological score and large intestine weight and length were estimated; mRNA expression of TNF-α, IL-1β, IL-6, IL-10 by quantitative PCR; VDR expression was detected by quantitative PCR and immunohistochemistry. In vitro: KLD restores epithelial cell-cell adhesion and mucosal healing during inflammation, while decreases the adhesiveness and invasiveness of AIEC bacteria and TNF-α and IL-8 mRNA expression and increases VDR expression. In vivo: KLD significantly improves body weight, clinical score, histological score and large intestine length of mice with DSS-induced colitis and reduces TNF-α, IL-1β and IL-6 mRNA levels, while increases IL-10 mRNA and VDR levels. KLD has significant effects on the intestinal mucosa, strongly decreasing inflammation, increasing epithelial restitution and reducing pathogenicity of harmful commensal bacteria.
目前，针对肠道炎症的原创治疗方法的研究集中在非药物治疗上。KLD 是磷虾油（KO）、益生菌罗伊氏乳杆菌（LR）和维生素 D（VitD3）的混合物。本研究旨在评估 KLD 在体外和体内减轻肠道炎症的潜在协同作用。分别使用结肠腺癌细胞系 CACO2 和 HT29 以及 C57BL/6 小鼠进行体外和体内分析。细胞暴露于细胞因子混合物（干扰素 γ+肿瘤坏死因子-α（TNF-α））以诱导炎症，或与 KO、LR 和 VitD3 单独或与细胞因子混合物和 KLD 共同暴露。动物用葡聚糖硫酸钠（DSS）处理 7 天以诱导结肠炎或用 DSS 和 KLD 处理。体外试验：通过免疫荧光分析 F-肌动蛋白表达；划痕试验和跨上皮电阻试验用于测量伤口愈合；黏附/侵袭试验检测黏附性和侵袭性大肠杆菌（AIEC）细菌；定量 PCR 检测 TNF-α、白细胞介素（IL）-8 和维生素 D 受体（VDR）的 mRNA 表达。体内试验：估计体重、临床评分、组织学评分和大肠重量和长度；定量 PCR 检测 TNF-α、IL-1β、IL-6、IL-10 的 mRNA 表达；定量 PCR 和免疫组织化学检测 VDR 表达。体外：KLD 在炎症过程中恢复上皮细胞-细胞黏附性和黏膜愈合，同时降低 AIEC 细菌的黏附性和侵袭性以及 TNF-α和 IL-8 mRNA 表达，增加 VDR 表达。体内：KLD 可显著改善 DSS 诱导的结肠炎小鼠的体重、临床评分、组织学评分和大肠长度，并降低 TNF-α、IL-1β 和 IL-6 mRNA 水平，同时增加 IL-10 mRNA 和 VDR 水平。KLD 对肠道黏膜有显著作用，强烈降低炎症，增加上皮修复，并减少有害共生菌的致病性。
