Vitamin D3 mediates autophagy to alleviate inflammatory responses in bovine endometrial epithelial cells and organoids via the PI3K/AKT/mTOR pathway

IF 1.4 3区 农林科学 Q4 IMMUNOLOGY Veterinary immunology and immunopathology Pub Date : 2024-10-11 DOI:10.1016/j.vetimm.2024.110839
Yalin Zhang, Xiaoyu Xie , Mingzhu Sun, Yujie Zhuang, Jin Zhou, Juanjuan Li, Penghui Yan, Juntao Zhang , Zhiping Zhang
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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.
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维生素 D3 通过 PI3K/AKT/mTOR 通路介导自噬,减轻牛子宫内膜上皮细胞和器官组织的炎症反应
作为一种天然抗炎剂,VD3(1,25 二羟基维生素 D3)的抗炎作用是否与自噬有关仍不清楚。本研究探讨了 VD3 对牛子宫内膜上皮细胞(BEECs)和牛子宫内膜器官组织(BEOs)的炎症损伤、自噬、氧化应激和细胞凋亡的影响。用 LPS(1 μg/ml)处理 BEECs 和 BEOs 24 小时,然后用 LPS+VD3 (50 ng/ml)处理 6 小时。使用 CCK8 检测法评估细胞活力。利用 qRT-PCR 和 Western 印迹分析量化了炎症因子(IL-1β、IL-6、TLR4、NF-κB)、自噬标记物(Beclin-1、ATG5、ATG7、p62)和 PI3K/AKT/mTOR 通路成分(PI3K、AKT 和 mTOR)的表达水平。结果表明,在 LPS+VD3 组中,IL-1β、IL-6、TLR4 和 NF-κB 的表达水平显著下降,CAT 和 SOD2 的活性明显提高(P < 0.05)。在 LPS+VD3 组中,自噬相关因子的表达明显增加,而信号通路因子的表达减少(P < 0.05)。此外,LPS+VD3 组细胞凋亡明显减少(P <0.05)。总之,这些研究结果表明,VD3能调节自噬,减轻BEECs和BEOs的氧化应激和炎症损伤,并通过PI3K/AKT/mTOR途径抑制LPS诱导的细胞凋亡。
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来源期刊
CiteScore
3.40
自引率
5.60%
发文量
79
审稿时长
70 days
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.
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