Wu Hongya, Fan Linlin, Gao Yanan, Wang Jiaqi, Zheng Nan
Key Laboratory of Quality & Safety Control for Milk and Dairy Products of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Laboratory of Quality and Safety Risk Assessment for Dairy Products of Ministry of Agriculture and Rural Affairs, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Foods. 2022 Oct 26;11(21):3378. doi: 10.3390/foods11213378.
Research evidence shows that effective nutritional intervention could prevent or reduce intestinal inflammatory injury in newborn infants. Iron free lactoferrin (apo-LF), one of the main types of lactoferrin (LF), is a bioactive protein in milk that plays a vital role in maintaining intestinal health. The potential mechanism by which apo-LF modulates intestinal inflammation is, however, still unclear. In the study we first explored key genes and pathways in vitro by transcriptome date analysis and then validated them in vivo to reveal the underlying molecular mechanism. The results showed that apo-LF pretreatment effectively inhibited lipopolysaccharide (LPS)-induced primary intestinal epithelial cells (IECs) inflammation in the co-culture system (primary IECs and immune cells), which was specifically manifested as the reduction of the concentration of TNF-α, IL-6 and IL-1β and increased the concentration of IFN-γ. In addition, transcriptome data analysis revealed that the key pathway for apo-LF to exert anti-inflammatory effects was the NF-κB/PPAR signaling pathway. Further validation was performed using western blotting in colonic tissues of young mice and it was found that the major proteins of NF-κB signaling pathway (NF-κB, TNF-α and IL-1β) were inhibited by apo-LF and the target proteins of PPAR signaling pathway (PPAR-γ and PFKFB3) were activated by apo-LF. Taken together, this suggests that apo-LF has a protective effect against LPS-induced intestinal inflammatory injury via modulating the NF-κB/PPAR signaling pathway, which provides new insights for further anti-inflammatory study of apo-LF.
研究证据表明,有效的营养干预可预防或减轻新生儿的肠道炎性损伤。乳铁蛋白主要类型之一的脱铁乳铁蛋白(apo-LF)是牛奶中的一种生物活性蛋白,在维持肠道健康方面发挥着至关重要的作用。然而,apo-LF调节肠道炎症的潜在机制仍不清楚。在本研究中,我们首先通过转录组数据分析在体外探索关键基因和信号通路,然后在体内进行验证,以揭示其潜在的分子机制。结果表明,apo-LF预处理在共培养系统(原代肠上皮细胞和免疫细胞)中有效抑制了脂多糖(LPS)诱导的原代肠上皮细胞(IECs)炎症,具体表现为肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)浓度降低,干扰素-γ(IFN-γ)浓度升高。此外,转录组数据分析显示,apo-LF发挥抗炎作用的关键信号通路是核因子-κB/过氧化物酶体增殖物激活受体(NF-κB/PPAR)信号通路。在幼鼠结肠组织中使用蛋白质免疫印迹法进行进一步验证,发现NF-κB信号通路的主要蛋白(NF-κB、TNF-α和IL-1β)受到apo-LF的抑制,PPAR信号通路的靶蛋白(PPAR-γ和6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3,PFKFB3)被apo-LF激活。综上所述,这表明apo-LF通过调节NF-κB/PPAR信号通路对LPS诱导的肠道炎性损伤具有保护作用,这为apo-LF的进一步抗炎研究提供了新的见解。