抵抗素通过激活AMPK/mTOR信号通路和自噬减轻脂多糖诱导的牛肺泡巨噬细胞炎症。
Resistin alleviates lipopolysaccharide-induced inflammation in bovine alveolar macrophages by activating the AMPK/mTOR signaling pathway and autophagy.
作者信息
Ma Xiaoping, Yang Aining, Fan Xiaoben, Liu Hong, Gu Yu, Wang Zhisheng, Guo Hongrui, Fang Jing, Cui Hengmin, Gou Liping, Deng Junliang, Cai Dongjie, Zuo Zhicai
机构信息
College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
College of Life Sciences, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
出版信息
Heliyon. 2024 Sep 18;10(19):e38026. doi: 10.1016/j.heliyon.2024.e38026. eCollection 2024 Oct 15.
OBJECTIVE
Resistin (RETN) is an adipocyte-specific hormone that participates in metabolism and modulates cellular inflammation. Our study aimed to assess the effects of RETN treatment on autophagy and the underlying molecular and biological mechanisms in bovine alveolar macrophages (BAMs).
METHODS
The optimal concentration of RETN + lipopolysaccharide (LPS) on macrophages was screened and then used to co-culture with alveolar macrophages. Autophagosomes in BAMs were examined using a transmission electron microscope (TEM). Quantitative real-time PCR (qRT-PCR) was used to detect the mRNA expression of microtubule-associated protein light chain 3 (LC3) and p62. Western blot (WB) was used to detect the protein expressions of LC3 and p62. The distribution of LC3 and p62 proteins in the cells was observed by immunofluorescence (IF). The concentrations of interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) were detected using enzyme-linked immunosorbent assay (ELISA). The protein expression of adenosine-monophosphate-activated protein kinase (AMPK), p-AMPK, mammalian target of rapamycin (mTOR), and p-mTOR was detected using WB.
RESULTS
The treatment of BAMs with RETN or LPS increased the number of autophagosomes and the ratio of LC3II/LC3I and decreased the expression level of p62 protein. RETN treatment significantly triggered autophagy compared to LPS treatment. Moreover, the ratios of p-AMPK/AMPK and p-mTOR/mTOR were upregulated and downregulated, respectively, after RETN treatment, suggesting that AMPK/mTOR signaling pathway activation is required for RETN-mediated autophagy in BAMs. Additionally, the ratio of LC3-II/LC3-I was lower, and the concentrations of IL-1β, IL-6, and TNF-α significantly decreased in the LPS and RETN co-treatment groups compared to the single LPS treatment group. However, both autophagy- and LPS-induced inflammation were partially alleviated by RETN treatment.
CONCLUSION
RETN can promote autophagy in BAMs by activating the AMPK/mTOR signaling pathway, it may help prevent LPS-induced inflammation.
目的
抵抗素（RETN）是一种脂肪细胞特异性激素，参与新陈代谢并调节细胞炎症。我们的研究旨在评估RETN处理对牛肺泡巨噬细胞（BAM）自噬的影响及其潜在的分子和生物学机制。
方法
筛选RETN+脂多糖（LPS）对巨噬细胞的最佳浓度，然后用于与肺泡巨噬细胞共培养。使用透射电子显微镜（TEM）检查BAM中的自噬体。采用定量实时PCR（qRT-PCR）检测微管相关蛋白轻链3（LC3）和p62的mRNA表达。使用蛋白质免疫印迹法（WB）检测LC3和p62的蛋白表达。通过免疫荧光（IF）观察LC3和p62蛋白在细胞中的分布。使用酶联免疫吸附测定（ELISA）检测白细胞介素（IL）-1β、IL-6和肿瘤坏死因子-α（TNF-α）的浓度。使用WB检测腺苷单磷酸活化蛋白激酶（AMPK）、p-AMPK、雷帕霉素靶蛋白（mTOR）和p-mTOR的蛋白表达。
结果
用RETN或LPS处理BAM可增加自噬体数量和LC3II/LC3I比率，并降低p62蛋白表达水平。与LPS处理相比，RETN处理显著触发自噬。此外，RETN处理后，p-AMPK/AMPK和p-mTOR/mTOR比率分别上调和下调，表明AMPK/mTOR信号通路激活是RETN介导BAM自噬所必需的。此外，与单一LPS处理组相比，LPS和RETN联合处理组中LC3-II/LC3-I比率较低，IL-1β、IL-6和TNF-α浓度显著降低。然而，RETN处理可部分减轻自噬和LPS诱导的炎症。
结论
RETN可通过激活AMPK/mTOR信号通路促进BAM自噬，可能有助于预防LPS诱导的炎症。
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