通过多种方法了解牛乳腺上皮细胞对急性热休克的早期反应

IF 4 2区 农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE Animal Pub Date : 2024-07-18 DOI:10.1016/j.animal.2024.101264
S. Castellani , C. Evangelista , M. Lepore , M. Portaccio , L. Basiricò , U. Bernabucci , I. Delfino
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引用次数: 0

摘要

热应激是奶牛场面临的一项重大挑战,会影响牛奶产量和质量,并在细胞水平上产生重要变化。有关热休克(HS)对奶牛乳腺细胞影响的体外研究大多侧重于中长期影响。近年来,傅立叶变换红外(FT-IR)显微光谱技术被越来越多地用于研究多种外部应激对不同细胞系的影响,直至单个细胞成分(如 DNA/RNA、脂质和蛋白质)的水平。本研究对牛乳腺上皮细胞(BME-UV1)在急性 HS(30 分钟-2 小时)诱导下可能发生的生化和分子水平变化进行了研究。将细胞暴露于不同的温度,即中性温度(TN,37 °C)和 HS 温度(42 °C),并获取傅立叶变换红外光谱,以分析 HS 对 BME-UV1 细胞成分(蛋白质、脂类和 DNA/RNA)的生化特性的影响。此外,还通过 RT-qPCR 分析了细胞活力测定、活性氧生成、热休克蛋白(HSPA1A、HSP90AA1、GRP78、GRP94)和抗氧化基因(SOD1、SOD2)的 mRNA 表达。傅立叶变换红外光谱(FT-IR)结果显示,与 TN 细胞相比,HS 细胞在接触 HS 30 分钟后,长链脂肪酸的含量就发生了变化,这可能是对 HS 细胞膜流动性改变的反应。暴露于 HS 2 小时后,HS 细胞中 DNA/RNA 活性的改变和聚集蛋白的积累凸显出来。基因表达分析表明,与 TN 细胞相比,HS 细胞中的 HSPA1A 和 HSP90AA1 在 30 分钟至 2 小时内过度表达。在接触 HS 2 小时后,HS 细胞中的 GRP94 也出现了过表达。急性 HS 并不影响 BME-UV1 细胞的活力、活性氧水平以及 SOD1 和 SOD2 基因的表达。根据研究结果,细胞在急性HS情况下会启动早期防御机制,这种高效的反应能力可能是奶牛耐受热应激的决定性因素。
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Insights on early response to acute heat shock of bovine mammary epithelial cells through a multimethod approach

Heat stress is a significant challenge in dairy cattle herds, affecting milk production and quality, and generating important changes at the cellular level. Most in vitro research on heat shock (HS) effects on dairy cow mammary cells was focused on medium-long-term effects. In recent years, Fourier transform-infrared (FT-IR) micro-spectroscopy has been increasingly used to study the effects of several external stresses on different cell lines, down to the level of single cellular components, such as DNA/RNA, lipids, and proteins. In this study, the possible changes at the biochemical and molecular level induced by acute (30 min-2 h) HS in bovine mammary epithelial (BME-UV1) cells were investigated. The cells were exposed to different temperatures, thermoneutral (TN, 37 °C) and HS (42 °C), and FT-IR spectra were acquired to analyse the effects of HS on biochemical characteristics of BME-UV1 cellular components (proteins, lipids, and DNA/RNA). Moreover, cell viability assay, reactive oxygen species production, and mRNA expression of heat shock proteins (HSPA1A, HSP90AA1, GRP78, GRP94) and antioxidant genes (SOD1, SOD2) by RT-qPCR were also analysed. The FT-IR results showed a change already at 30 min of HS exposure, in the content of long-chain fatty acids, which probably acted as a response to a modification of membrane fluidity in HS cells compared with TN cells. After 2 h of HS exposure, modification of DNA/RNA activity and accumulation of aggregated proteins was highlighted in HS cells. The gene expression analyses showed the overexpression of HSPA1A and HSP90AA1 starting from 30 min up to 2 h in HS cells compared with TN cells. At 2 h of HS exposure, also the overexpression of GRP94 was observed in HS cells. Acute HS did not affect cell viability, reactive oxygen species level, and SOD1 and SOD2 gene expression of BME-UV1 cells. According to the results obtained, cells initiate early defence mechanisms in case of acute HS and probably this efficient response capacity may be decisive for tolerance to heat stress of dairy cattle.

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来源期刊
Animal
Animal 农林科学-奶制品与动物科学
CiteScore
7.50
自引率
2.80%
发文量
246
审稿时长
3 months
期刊介绍: Editorial board animal attracts the best research in animal biology and animal systems from across the spectrum of the agricultural, biomedical, and environmental sciences. It is the central element in an exciting collaboration between the British Society of Animal Science (BSAS), Institut National de la Recherche Agronomique (INRA) and the European Federation of Animal Science (EAAP) and represents a merging of three scientific journals: Animal Science; Animal Research; Reproduction, Nutrition, Development. animal publishes original cutting-edge research, ''hot'' topics and horizon-scanning reviews on animal-related aspects of the life sciences at the molecular, cellular, organ, whole animal and production system levels. The main subject areas include: breeding and genetics; nutrition; physiology and functional biology of systems; behaviour, health and welfare; farming systems, environmental impact and climate change; product quality, human health and well-being. Animal models and papers dealing with the integration of research between these topics and their impact on the environment and people are particularly welcome.
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