Mohsin Muhammad Ali, Zhou Xiaojing, Huiru Yu, Shen Wenxiang, He Baoxiang, Sobiech Przemysław, Pierzchała Mariusz, Ogłuszka Magdalena, Starzyński Rafał, Kalra Garima, Deshmukh Bharti, Thangarasu Revathy, Kashyap Neeraj, Czarnik Urszula, Lepczyński Adam, Woźniakowski Grzegorz, Pareek Chandra S
Department of Clinical Veterinary Medicine, College of Animal Science and Technology, Guangxi University, Nanning, 530004, Guangxi, China.
Shanghai Animal Disease Prevention and Control Center, Changning District, 201103, Shanghai, China.
J Vet Res. 2024 Jun 22;68(2):313-324. doi: 10.2478/jvetres-2024-0037. eCollection 2024 Jun.
In dairy cattle, oxidative stress is a predominant problem associated with diseases and reproductive health issues. This study aimed to detect the variation in the antioxidant biomarkers by adding different concentrations of β-hydroxybutyric acid (BHBA) and sought to elucidate its effects on the gene expression levels of growth hormone (GH) and antioxidant biomarkers in bovine hepatocytes.
Four antioxidant biomarkers, namely malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH Px) were evaluated using commercially available bovine ELISA kits. The expression levels of the bovine GH, its receptor (GHR), insulin-like growth factor (IGF), IGF-1, IGF-1 receptor, CAT, SOD, GSH-Px and β-actin (as a reference) genes in liver cell culture were determined by reverse transcriptase-PCR assay.
With the increase of BHBA concentration and culture time, the activities of SOD, CAT, and GSH Px biomarkers in hepatocytes decreased. However, the content of MDA in hepatocytes increased gradually with the increase of hepatocyte culture time and BHBA concentration. The qPCR results revealed that after adding BHBA, gene expression levels of GSH-Px, SOD and IGF biomarkers in hepatocytes began to differ in the culture groups at 12 h, whereas the gene expression level of the CAT and GHR biomarkers in hepatocytes began to differ at 6 h.
Quantitative PCR results showed that the BHBA significantly downregulated the expression levels of the GHR gene and CAT, GSH Px and SOD antioxidant biomarker genes.
在奶牛中,氧化应激是与疾病和生殖健康问题相关的一个主要问题。本研究旨在通过添加不同浓度的β-羟基丁酸(BHBA)来检测抗氧化生物标志物的变化,并试图阐明其对牛肝细胞中生长激素(GH)基因表达水平和抗氧化生物标志物的影响。
使用市售的牛ELISA试剂盒评估四种抗氧化生物标志物,即丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH Px)。通过逆转录-PCR测定法测定肝细胞培养物中牛GH、其受体(GHR)、胰岛素样生长因子(IGF)、IGF-1、IGF-1受体、CAT、SOD、GSH-Px和β-肌动蛋白(作为对照)基因的表达水平。
随着BHBA浓度和培养时间的增加,肝细胞中SOD、CAT和GSH Px生物标志物的活性降低。然而,肝细胞中MDA的含量随着肝细胞培养时间和BHBA浓度的增加而逐渐增加。qPCR结果显示,添加BHBA后,肝细胞中GSH-Px、SOD和IGF生物标志物的基因表达水平在培养12小时时开始在各培养组中出现差异,而肝细胞中CAT和GHR生物标志物的基因表达水平在6小时时开始出现差异。
定量PCR结果表明,BHBA显著下调了GHR基因以及CAT、GSH Px和SOD抗氧化生物标志物基因的表达水平。