Šeškutė Milda, Laucaitytė Goda, Inčiūraitė Rūta, Malinauskas Mantas, Jankauskaitė Lina
Department of Pediatrics, Lithuanian University of Health Sciences, 44307 Kaunas, Lithuania.
Institute of Physiology and Pharmacology, Lithuanian University of Health Sciences, 44307 Kaunas, Lithuania.
Cells. 2025 Jun 28;14(13):990. doi: 10.3390/cells14130990.
The gastrointestinal (GI) tract is a major barrier against pathogens, including viruses. The antiviral responses in the GI epithelium have been broadly investigated, but data on the contribution of the stromal cells remain scarce. Melatonin, widely used to treat insomnia, has recently been proposed as an antiviral agent, yet its effect in the GI tract remains poorly understood. We compared the antiviral responses in Caco-2 monocultures and co-cultures with intestinal fibroblasts (HSIFs) and endothelial cells (HUVECs) after stimulation using Poly I:C. We evaluated the apoptosis, proliferation, key antiviral markers (IRF1, IRF3, IFNs, TBK1, STAT3), and mitochondrial and peroxisomal activation with and without melatonin. The Caco-2 cells cultured with the HSIFs and HUVECs demonstrated enhanced proliferation and reduced Poly I:C-induced apoptosis. The co-culture exhibited a more rapid IRF3-IFNλ1 response, higher TBK1 expression, and enhanced peroxisomal activation compared to these properties in the monoculture. Melatonin further reduced apoptosis and modulated organelle-specific antiviral signaling by suppressing peroxisomal activation and promoting mitochondrial activity. Reduced peroxisomal activation was associated with decreased TBK1, IRF3, and IFNλ1 levels and altered STAT3 signaling. These effects were more pronounced when melatonin was applied post-stimulation compared to that under prophylactic use. Fibroblast-endothelial interactions amplify the antiviral responses in the intestinal epithelial cells by activating the TBK1-IRF3-IFNλ1 axis. Melatonin modulates these responses, highlighting its therapeutic potential in viral GI infections.
胃肠道（GI）是抵御包括病毒在内的病原体的主要屏障。人们已经广泛研究了胃肠道上皮中的抗病毒反应，但关于基质细胞作用的数据仍然很少。褪黑素广泛用于治疗失眠，最近被提议作为一种抗病毒药物，但其在胃肠道中的作用仍知之甚少。我们比较了使用聚肌胞苷酸（Poly I:C）刺激后，Caco-2单培养物以及与肠道成纤维细胞（HSIFs）和内皮细胞（HUVECs）共培养物中的抗病毒反应。我们评估了有无褪黑素情况下的细胞凋亡、增殖、关键抗病毒标志物（IRF1、IRF3、干扰素、TBK1、STAT3）以及线粒体和过氧化物酶体的激活情况。与HSIFs和HUVECs共培养的Caco-2细胞表现出增殖增强以及聚肌胞苷酸诱导的细胞凋亡减少。与单培养物相比，共培养物表现出更快的IRF3 - 干扰素λ1反应、更高的TBK1表达以及增强的过氧化物酶体激活。褪黑素通过抑制过氧化物酶体激活和促进线粒体活性，进一步减少细胞凋亡并调节细胞器特异性抗病毒信号传导。过氧化物酶体激活的减少与TBK1、IRF3和干扰素λ1水平的降低以及STAT3信号传导的改变有关。与预防性使用相比，刺激后应用褪黑素时这些作用更为明显。成纤维细胞 - 内皮细胞相互作用通过激活TBK1 - IRF3 - 干扰素λ1轴增强肠道上皮细胞中的抗病毒反应。褪黑素调节这些反应，突出了其在病毒性胃肠道感染中的治疗潜力。
