[miR-373 inhibits M2 polarization of tumor associated macrophages and affects rectal cancer cells by regulating JAK2/STAT6 signal pathway]Li, Wu, Xie
et alXi Bao Yu Fen Zi Mian Yi Xue Za Zhi (2025) 41 (3), 211-220
Abstract: Objective To explore the effects of miR-373 and Janus kinase 2/signal transducer and activator of transcription 6 (JAK2/STAT6) signaling pathways on the M2 polarization of tumor associated macrophages (TAM) in rectal cancer. Methods THP-1 cells were induced into M0/M1/M2 macrophages, M0 macrophages were cocultured with Caco-2 cells to obtain TAM, Flow cytometry was used to detect the expression of CD86 and CD206, Real-time quantitative qPCR and Western blot were used to detect miR-373, inducible nitric oxide synthase (iNOS), toll-like receptor 4 (TLR-4), interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), arginase 1 (Arg1), chitinase 3-like 1 (Ym1), resistin like α (Fizz1), IL-10 mRNA and protein levels. TAM were transfected and divided into overexpressing miR-373 group (miR-373-TAM) and control group (miR-NC-TAM), overexpressing miR-373+JAK2-TAM group (miR-373 combined with JAK2-TAM) and control group (miR-373 combined with NC-TAM), and then cocultured with Caco-2 cells. Flow cytometry was used to detect the expression of CD206 in TAM; Real-time quantitative PCR and Western blot were used to detect miR-373, Arg1, Ym1, Fizz1, IL-10, JAK2, STAT6 mRNA and protein levels in TAM; CCK-8 assay, colony formation assay, and Transwell assay were used to detect the proliferation, migration, and invasion ability of Caco-2 cells. Thirty nude mice were randomly divided into Caco-2 cells group, Caco-2 cells combined with miR-NC-TAM group, and Caco-2 cells combined with miR-373-TAM group, with 10 mice in each group. Rats in each group were subcutaneously injected with pure Caco-2 cells, Caco-2 cells combined with TAM, and Caco-2 cells combined with TAM overexpressing miR-373. After 4 weeks of cell inoculation, immunofluorescence staining was used to detect F4/80+CD206+cells level in tumor tissue; Real-time quantitative PCR and Western blot were used to detect miR-373, JAK2, STAT6, Arg1, Ym1, Fizz1, IL-10 mRNA and protein levels in tumor tissues. Results TAM tended to M2 polarization. After overexpression of miR-373, miR-373 level in TAM was increased, while Arg1, Ym1, Fizz1, IL-10, JAK2, STAT6 mRNA and protein levels were decreased, proliferation, migration, invasion ability of Caco-2 cells were decreased; Overexpression of JAK2 could partially reverse the effect of overexpression of miR-373 on the M2 polarization of TAM and proliferation, migration, invasion ability of Caco-2 cells. TAM could promote tumor growth; Overexpression of miR-373 could inhibit tumor growth and inhibit M2 polarization of TAM. Conclusion miR-373 could inhibit M2 polarization of TAM in rectal cancer, and miR-373 might inhibit proliferation and metastasis of rectal cancer cells by regulating the JAK2/STAT6 pathway.
Features of toll-like receptor genes (TLR-2, TLR-3, TLR-4 and TLR-6) polymorphism in open-angle glaucoma patientsShevchenko, Prokofiev, Konenkov
et alVavilovskii Zhurnal Genet Selektsii (2025) 29 (1), 128-134
Abstract: Modern research shows that innate immunity plays an important role in the pathogenesis of primary open-angle glaucoma (POAG). An increase in the content of toll-like receptors (TLR) in the glaucomatous retina of the human eye was revealed. TLRs can modulate the immune response in glaucoma; provide early recognition of damaging agents, activation of signaling pathways and effector mechanisms of the nonspecific immune defense system aimed at restoring homeostasis. The TLR-encoding genes' polymorphism alters the amino acid structure of the receptors, which leads to changes in their immune functions: expression level, ligand-binding and coreceptor functions, transport and signal transmission. The aim was to analyze the association of the TLR2 (rs5743708), TLR3 (rs3775291), TLR4 (rs4986790, rs4986791) and TLR6 (rs5743810) polymorphisms with primary open-angle glaucoma in patients of Western Siberia.99 patients (52 men and 47 women) with a diagnosis of primary open-angle glaucoma were examined. The comparison group consisted of 100 people (81 women and 19 men). TLR2 (rs5743708), TLR3 (rs3775291), TLR4 (rs4986790, rs4986791) and TLR6 (rs5743810) polymorphisms were analyzed by RT-PCR using test systems with Syber Green (Lytex, Russia). Statistical analysis was performed using the software package SPSS 23.0 and Arlequin 3.5.2.2.the distribution of genotypes in the patient group and in the control group corresponded to the Hardy-Weinberg equilibrium. The genotype frequencies did not significantly differ between the two analyzed groups. The frequency of TLR2-753 ArgArg:TLR6-249 ProPro was increased in the group of patients with POAG. The linkage disequilibrium between two polymorphic positions of the TLR4 gene was revealed. In addition, the linkage disequilibrium between TLR2-TLR6 gene for the glaucoma group and the control group was revealed.an increase in certain genotypes in the patient group relative to the control group may indirectly indicate the involvement of infectious factors in the initiation of POAG. However, despite the proven importance of the participation of their protein products in the pathogenesis of glaucoma, the relationship of TLR polymorphism requires additional research taking into account the ethnic characteristics of patients and intergenic interactions for a better understanding of the complex mechanisms of disease development. This will help carry out early diagnosis and develop the necessary therapeutic strategy.Copyright © AUTHORS.
Effects of lipopolysaccharide administration on thymus damage, antioxidant capacity and immune function in weaned pigletsBai, Jiang, Li
et alJ Vet Res (2025) 69 (1), 111-119
Abstract: Piglets are vulnerable to stress during weaning because of changes in the feeding environment, nutrients, and other growth-impacting conditions. In this study, stress injury was modelled by continuous intraperitoneal injection of lipopolysaccharide (LPS) and was used to investigate the dynamics of antioxidant indices and immunoinflammatory factors in the piglet thymus.Forty-eight weaned piglets were divided into an LPS group and a control group. One group was injected with LPS solution (100 μg/kg) and the other with sterile saline daily. The experiment ran over 13 days, and six piglets from each group were euthanised for necropsy on days 1, 5, 9 and 13. Thymic tissues were collected, and the antioxidant indices and mRNA expression levels of related genes were measured by enzyme activity assay and reverse-transcription quantitative PCR.In the LPS group, catalase activities were significantly increased on days 1 and 5, that of superoxide dismutase was significantly higher on day 9 and glutathione activity was elevated throughout. Messenger RNA (mRNA) expression of the toll-like receptor 4 (TLR4) pathway, interleukin (IL) 6, and IL-2 increased in the thymus on day 1. By day 5, the mRNA expression of the TLR pathway, the janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway, the kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, tumour necrosis factor α, IL-10, IL-6 and IL-2 were decreased. On day 13, the mRNA expression levels of the TLR4 and Keap1/Nrf2 pathways, TNF-α, IL-10 and IL-6 increased again.Continuous LPS induction led to high activation of the thymic immune system in piglets during the prophase. However, this activation was accompanied by atrophy and immunosuppression mid-experiment. Nevertheless, the immune function gradually recovered in the later stages.© 2025 Lingna Bai et al., published by Sciendo.
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