摘要
为探究围产期奶牛亚临床酮病血清中生化指标的变化特征,本研究选取2~4胎次荷斯坦围产期奶牛45头,分别在分娩前7 d、分娩当天晨饲前经尾根静脉采血,以血酮检测值<1.2 mmol/L为健康组,≥1.2 mmol/L为酮病组。采用ELISA试剂盒测定分娩前7 d、分娩当天、分娩后7 d、14 d健康组和酮病组奶牛血液中的生化指标。结果显示,酮病组奶牛分娩前7 d和分娩当天血清中皮质醇含量显著高于健康组(P<0.05);并且酮病组奶牛分娩前7 d血清中β-羟丁酸(BHBA)、高密度脂蛋白(HDL-C)含量显著高于健康组(P<0.05);酮病组奶牛分娩前后血清中葡萄糖(Glu)、甘油三脂(TG)、极低密度脂蛋白(VLDL-C)、游离脂肪酸(FFA)含量较健康组虽无显著差异,但均高于健康组。分别以0、1×10^(-7)mmol/L、1×10^(-8)mmol/L、1×10^(-9)mmol/L浓度的糖皮质激素(GC)刺激原代分离培养的犊牛肝细胞,作用0、1 h、3 h、6 h、24 h后采用荧光定量PCR检测肝细胞中糖异生相关受体及其相关酶编码基因的相对转录水平,并以皮尔森相关系数对糖异生相关受体及其相关酶编码基因的转录水平进行相关性分析。q PCR结果显示:高浓度GC促进原代犊牛肝细胞中糖皮质激素受体(GR)NR3CI的转录水平,低浓度GC促进盐皮质激素受体(MR)NR3C2的转录水平。糖异生相关限速酶PC、PEPCK-C、PCK2、11β-HSD1、11β-HSD2编码基因的相对转录水平随GC浓度和其作用时间而变化。皮尔森(Pearson)相关性分析结果显示:PC、PCK2、11β-HSD1基因的相对转录水平与NR3C1极显著相关(P<0.01),PEPCK-C、11β-HSD1、11β-HSD2与NR3C2的相对转录水平显著相关(P<0.05)。上述结果表明酮病奶牛分娩前GC水平升高,而GC可能通过结合其受体GR/MR提高奶牛肝细胞中糖异生关键酶PC、PEPCK-C、PCK2、11β-HSD1和11β-HSD2编码基因m RNA的转录,从而促进糖异生作用。本研究为酮病奶牛体内糖脂代谢相关激素GC的内分泌调节作用�
The aim of this study was to explore the changes of serum biochemical indexes of perinatal dairy cows with subclinical ketosis,analyze the changes of the level of glucocorticoid cortisol(Cor),and to investigate the molecular mechanism of synthetic glucocorticoid(GC)regulating gluconeogenesis in liver cells of dairy cows by primary isolation of calf hepatocytes.45 Holstein perinatal cows of 2-4 parities were selected,and blood was collected from the anterior caudal vein 7 days before delivery and in the morning of delivery.The BHBCheck Plus Blood Ketone blood ketone test value<1.2 mmol/L was used as the healthy group,and those value≥1.2 mmol/L were used as the ketosis group.ELISA method was used to measure biochemical indexes in the blood of cows in the healthy group and ketosis group at 0 and 7 days before delivery,7 days and 14 days after delivery,respectively.The results showed that the serum cortisol content in the ketosis group was significantly higher than that in the healthy group 7 days before delivery and on the day of delivery(P<0.05).The contents ofβ-hydroxybutyric(BHBA)and high-density lipoprotein(HDL-C)in the serum of cows in the ketosis group 7 days before delivery were significantly higher than those in the healthy group(P<0.05).The levels of glucose(Glu),triglycerides(TG),extremely low density lipoprotein(VLDL-C)and free fatty acids(FFA)in the serum of cows with ketosis before and after delivery were not significantly different from those in healthy group,but were all higher than those in healthy group.The primary cultured calf hepatocytes were further stimulated by glucocorticoids at concentrations of 0,1×10^(-7)mmol/L,1×10^(-8)mmol/L and 1×10^(-9)mmol/L,respectively.After 0,1 hours,3 hours,6 hours and 24 hours,the relative transcription levels of gluconeogenic receptors and genes in hepatocytes were detected by qPCR method,and the correlation between the expression levels of gluconeogenic receptors and genes was analyzed by Pearson correlation coefficient.The results showed that high concen
作者
陈灰
李松建
杜蕊
徐子萱
李薛强
CHEN Hui;LI Song-jian;DU Rui;XU Zi-xuan;LI Xue-qiang(Inner Mongolia Agricultural University,College of Veterinary Medicine,Hohhot 010018,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2023年第9期959-967,共9页
Chinese Journal of Preventive Veterinary Medicine
基金
内蒙古自然科学基金面上项目(2019MS03056)
内蒙古自然科学基金博士基金项目(2019BS03017)
内蒙古农业大学高层次人才科研启动项目(NDYB2018-10)。
关键词
奶牛酮病
糖皮质激素
糖异生
糖皮质激素受体
盐皮质激素受体
dairy cattle ketosis
cortisol
gluconeogenesis
glucocorticoid receptor(GR)
mineralocorticoid receptor(MR)
