{"title":"Vitamin D3 mediates autophagy to alleviate inflammatory responses in bovine endometrial epithelial cells and organoids via the PI3K/AKT/mTOR pathway","authors":"Yalin Zhang, Xiaoyu Xie , Mingzhu Sun, Yujie Zhuang, Jin Zhou, Juanjuan Li, Penghui Yan, Juntao Zhang , Zhiping Zhang","doi":"10.1016/j.vetimm.2024.110839","DOIUrl":null,"url":null,"abstract":"<div><div>As a natural anti-inflammatory agent, it remains unclear whether the anti-inflammatory effects of VD3 (1,25 dihydroxyvitamin D3) are related to autophagy. This study investigates the impact of VD3 on inflammatory injury, autophagy, oxidative stress, and apoptosis in bovine endometrial epithelial cells (BEECs) and bovine endometrial organoids (BEOs). BEECs and BEOs were treated with LPS (1 μg/ml) for 24 hours, followed by treatment with LPS+VD3 (50 ng/ml) for 6 hours. Cell viability was assessed using the CCK8 assay. The expression levels of inflammatory factors (IL-1β, IL-6, TLR4, NF-κB), autophagy markers (Beclin-1, ATG5, ATG7, p62), and components of the PI3K/AKT/mTOR pathway (PI3K, AKT, and mTOR) were quantified using qRT-PCR and Western blot analyses. LC3B expression was detected by immunofluorescence, and the apoptosis rate was assessed using Annexin V. The results demonstrated a significant decrease in the expression levels of IL-1β, IL-6, TLR4, and NF-κB, along with a notable increase in the activity of <em>CAT</em> and <em>SOD2</em> in the LPS+VD3 group (<em>P</em> < 0.05). The expression of autophagy-related factors was significantly increased, whereas the expression of signaling pathway factors was decreased in the LPS+VD3 group (<em>P</em> < 0.05). Additionally, apoptosis was significantly alleviated in the LPS+VD3 group (<em>P</em> < 0.05). Collectively, these findings indicate that VD3 modulates autophagy, attenuates oxidative stress and inflammatory damage in BEECs and BEOs, and inhibits LPS-induced apoptosis via the PI3K/AKT/mTOR pathway.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":null,"pages":null},"PeriodicalIF":1.4000,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary immunology and immunopathology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0165242724001259","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
As a natural anti-inflammatory agent, it remains unclear whether the anti-inflammatory effects of VD3 (1,25 dihydroxyvitamin D3) are related to autophagy. This study investigates the impact of VD3 on inflammatory injury, autophagy, oxidative stress, and apoptosis in bovine endometrial epithelial cells (BEECs) and bovine endometrial organoids (BEOs). BEECs and BEOs were treated with LPS (1 μg/ml) for 24 hours, followed by treatment with LPS+VD3 (50 ng/ml) for 6 hours. Cell viability was assessed using the CCK8 assay. The expression levels of inflammatory factors (IL-1β, IL-6, TLR4, NF-κB), autophagy markers (Beclin-1, ATG5, ATG7, p62), and components of the PI3K/AKT/mTOR pathway (PI3K, AKT, and mTOR) were quantified using qRT-PCR and Western blot analyses. LC3B expression was detected by immunofluorescence, and the apoptosis rate was assessed using Annexin V. The results demonstrated a significant decrease in the expression levels of IL-1β, IL-6, TLR4, and NF-κB, along with a notable increase in the activity of CAT and SOD2 in the LPS+VD3 group (P < 0.05). The expression of autophagy-related factors was significantly increased, whereas the expression of signaling pathway factors was decreased in the LPS+VD3 group (P < 0.05). Additionally, apoptosis was significantly alleviated in the LPS+VD3 group (P < 0.05). Collectively, these findings indicate that VD3 modulates autophagy, attenuates oxidative stress and inflammatory damage in BEECs and BEOs, and inhibits LPS-induced apoptosis via the PI3K/AKT/mTOR pathway.
期刊介绍:
The journal reports basic, comparative and clinical immunology as they pertain to the animal species designated here: livestock, poultry, and fish species that are major food animals and companion animals such as cats, dogs, horses and camels, and wildlife species that act as reservoirs for food, companion or human infectious diseases, or as models for human disease.
Rodent models of infectious diseases that are of importance in the animal species indicated above,when the disease requires a level of containment that is not readily available for larger animal experimentation (ABSL3), will be considered. Papers on rabbits, lizards, guinea pigs, badgers, armadillos, elephants, antelope, and buffalo will be reviewed if the research advances our fundamental understanding of immunology, or if they act as a reservoir of infectious disease for the primary animal species designated above, or for humans. Manuscripts employing other species will be reviewed if justified as fitting into the categories above.
The following topics are appropriate: biology of cells and mechanisms of the immune system, immunochemistry, immunodeficiencies, immunodiagnosis, immunogenetics, immunopathology, immunology of infectious disease and tumors, immunoprophylaxis including vaccine development and delivery, immunological aspects of pregnancy including passive immunity, autoimmuity, neuroimmunology, and transplanatation immunology. Manuscripts that describe new genes and development of tools such as monoclonal antibodies are also of interest when part of a larger biological study. Studies employing extracts or constituents (plant extracts, feed additives or microbiome) must be sufficiently defined to be reproduced in other laboratories and also provide evidence for possible mechanisms and not simply show an effect on the immune system.