Batt Mackenzie C, Gibbs Rachel L, Reith Rachel R, Fuller Anna M, Yates Dustin T, Petersen Jessica L
Department of Animal Science, University of Nebraska-Lincoln, Lincoln, NE 68583, USA.
J Anim Sci. 2025 Jan 4;103. doi: 10.1093/jas/skaf116.
Lidocaine is a commonly used local anesthetic that blocks sodium channels in nociceptor neurons, preventing the transmission of pain signals to the brain. Lidocaine can be administered to reduce discomfort during tissue biopsies. Biopsy tissue may then be used to study the transcriptome under the assumption that the genomic activity of lidocaine-treated tissue accurately reflects that of untreated tissue. This study investigated how intramuscular lidocaine injection influenced skeletal muscle gene expression in sheep, aiming to understand how transcriptomic changes could affect data interpretation. Approximately 10 min before euthanasia, the left biceps brachii muscle from each of 6 wether lambs (48.7 ± 0.8 kg) was injected (IM; 20G hypodermic needle) at a depth of 3 cm with 2 mL of 2% lidocaine (20 mg/mL); the right biceps brachii was untreated. At necropsy, muscle samples were collected from the injection sites and contralateral limbs and flash-frozen. In an additional set of lambs, lidocaine-treated and untreated samples were collected from the biceps brachii of 4 lambs, and the vastus intermedius of 4 other lambs. RNA was isolated and mRNA sequenced to a targeted depth of 20 million reads per sample. Sequences were mapped and quantified; matched-pair analysis was performed in EdgeR. No genes were consistently differentially expressed due to treatment in both muscle types, perhaps reflecting their distinct physiological roles. Lidocaine did influence the transcriptome with anti-inflammatory effects evident in both muscle types, including the downregulation of immune-associated transcription factors and other genes. Lidocaine's influence varied on other broad categories of genes, including those associated with muscle contractility, tissue repair, and structural integrity, which could affect the interpretation of transcriptome data in studies of muscle growth and development. Pathway analysis revealed that lidocaine impacted signaling mechanisms for cellular connectivity and structure. This study demonstrates that intramuscular administration of lidocaine results in the alteration of tissue's gene expression profiles, highlighting the importance of considering lidocaine's influence in transcriptome analyses. Thus, the use of complementary physiological measures to validate transcriptomic findings is recommended to ensure observed gene expression changes are accurately attributed to experimental conditions rather than the effects of lidocaine.
利多卡因是一种常用的局部麻醉剂，它能阻断伤害感受器神经元中的钠通道，从而阻止疼痛信号向大脑的传递。利多卡因可用于减轻组织活检过程中的不适。然后，活检组织可用于研究转录组，前提是利多卡因处理过的组织的基因组活性能准确反映未处理组织的基因组活性。本研究调查了肌肉注射利多卡因如何影响绵羊骨骼肌基因表达，旨在了解转录组变化如何影响数据解读。在安乐死大约10分钟前，给6只去势羔羊（体重48.7±0.8千克）的每只羊的左肱二头肌（肌肉注射；20G皮下注射针）在3厘米深处注射2毫升2%利多卡因（20毫克/毫升）；右肱二头肌未处理。尸检时，从注射部位和对侧肢体采集肌肉样本并速冻。在另一组羔羊中，从4只羔羊的肱二头肌以及另外4只羔羊的股中间肌采集利多卡因处理和未处理的样本。分离RNA并对mRNA进行测序，每个样本的目标深度为2000万条读数。对序列进行比对和定量；在EdgeR中进行配对分析。两种肌肉类型中均未发现因处理而持续差异表达的基因，这可能反映了它们不同的生理作用。利多卡因确实影响了转录组，在两种肌肉类型中均有明显的抗炎作用，包括免疫相关转录因子和其他基因的下调。利多卡因对其他广泛类别的基因的影响各不相同，包括与肌肉收缩性、组织修复和结构完整性相关的基因，这可能会影响肌肉生长和发育研究中转录组数据的解读。通路分析表明，利多卡因影响细胞连接和结构的信号传导机制。本研究表明，肌肉注射利多卡因会导致组织基因表达谱的改变，突出了在转录组分析中考虑利多卡因影响的重要性。因此，建议使用补充性生理指标来验证转录组学研究结果，以确保观察到的基因表达变化准确归因于实验条件而非利多卡因的影响。
