Kongara Kavitha, Dukkipati Venkata Sayoji Rao, Tai Hui Min, Heiser Axel, Murray Alan, Webster James, Johnson Craig Brian
School of Veterinary Science, Massey University, Palmerston North 4410, New Zealand.
School of Agriculture and Environment, Massey University, Palmerston North 4410, New Zealand.
Animals (Basel). 2020 Jul 14;10(7):1187. doi: 10.3390/ani10071187.
Calf disbudding is a painful husbandry practice on dairy and beef cattle farms. An objective measurement of pain is useful to reliably evaluate the pain intensity and anti-nociceptive (analgesic) efficacy of therapeutic agents. The aim of this study was to investigate the changes in peripheral leucocyte inflammatory cytokine gene expression in calves after disbudding, and to assess whether the changes in cytokine gene expression could be an indicator of the efficacy of analgesic drugs. In a randomised controlled study, 16 calves (aged 31 to 41 days and weighing 58 to 73 kg), undergoing routine disbudding, were randomly allocated into two groups ( = 8 in each group). Calves in the control group received no analgesic, while those in the treatment group received 0.5 mg kg meloxicam subcutaneously prior to disbudding. Disbudding was performed using an electric debudder. Blood (10 mL) was sampled from the jugular vein just before and 4 and 24 h post-disbudding, RNA was extracted from leukocytes, and the transcription of 12 genes of interest was assessed using nCounter gene expression assay. The results showed significantly higher transcription (compared to baseline values) of the studied genes (except , , and ) in the control group calves at either 4 or 24 h post-disbudding. The administration of meloxicam one hour before disbudding significantly attenuated the upregulation of , , , , and gene transcription post-disbudding, while it did not suppress the elevated transcription of acute and pro-inflammatory cytokines such as , , , and genes. In conclusion, nCounter gene expression assay seems to be a promising tool to study the expression of cytokine genes and thus could be used for the pre-clinical evaluation of novel analgesics.
犊牛去角是奶牛场和肉牛场中一项痛苦的饲养操作。客观测量疼痛对于可靠评估治疗药物的疼痛强度和抗伤害感受(镇痛)效果很有用。本研究的目的是调查去角后犊牛外周血白细胞炎性细胞因子基因表达的变化,并评估细胞因子基因表达的变化是否可作为镇痛药疗效的指标。在一项随机对照研究中,16头(年龄31至41天,体重58至73千克)接受常规去角的犊牛被随机分为两组(每组 = 8头)。对照组犊牛不接受任何镇痛药,而治疗组犊牛在去角前皮下注射0.5毫克/千克美洛昔康。使用电动去角器进行去角。在去角前、去角后4小时和24小时从颈静脉采集10毫升血液,从白细胞中提取RNA,并使用nCounter基因表达分析评估12个感兴趣基因的转录情况。结果显示,对照组犊牛在去角后4小时或24小时,所研究基因(除 、 和 外)的转录水平显著高于基线值。去角前1小时给予美洛昔康可显著减轻去角后 、 、 、 和 基因转录的上调,而它并未抑制 、 、 和 等急性和促炎细胞因子基因转录的升高。总之,nCounter基因表达分析似乎是研究细胞因子基因表达的一种有前景的工具,因此可用于新型镇痛药的临床前评估。