快型和慢型肉鸡肌纤维发育差异的 IGF1-PI3K/AKT 信号通路调控机制

Regulatory mechanisms of the IGF1-PI3K/AKT signaling pathway in myofiber developmental differences between fast- and slow-growing broilers

摘要

[目的]本文旨在探究不同生长速度肉鸡在肌纤维结构及其分子调控机制上的差异，揭示肌纤维发育特点及分子调控机制。[方法]选取快速生长的 Cobb500 肉鸡和慢速生长的皖南三黄鸡为研究对象，分别取 18 胚龄、21 日龄和 42 日龄的快、慢型肉鸡肌肉样本进行苏木精-伊红染色，通过扫描电镜观察肌纤维形态结构，利用实时荧光定量 RT-qPCR 和蛋白质免疫印迹检测肌纤维发育关键基因表达。构建胰岛素样生长因子 1(IGF1)敲除和过表达质粒并转染至鸡胚成纤维细胞(DF-1)，验证 IGF1 的作用通路及功能。[结果]快速型肉鸡 Cobb500 在 18 胚龄肌纤维直径显著低于慢速型肉鸡三黄鸡(P<0.05)，在 21 和 42 日龄显著高于慢速型肉鸡(P<0.05)。快速型肉鸡 18 胚龄的肌纤维密度显著高于慢速型肉鸡(P<0.05)，在 21 和 42 日龄显著低于慢速型肉鸡(P<0.05)。快速型肉鸡肌纤维中 IGF1 基因和蛋白表达量在 18 胚龄、21 日龄和 42 日龄显著高于慢速型肉鸡(P<0.05)。与阴性对照组和 IGF1 敲除组相比，过表达 IGF1 显著增加丝氨酸/苏氨酸激酶 1(AKT1)、细胞周期蛋白依赖性激酶 4(CDK4)和 c-Jun N 末端激酶(JNK)基因表达(P<0.05)。在质粒转染 16~40h，过表达 IGF1 质粒转染组细胞活力显著高于敲除 IGF1 质粒转染组和空白对照组(P<0.05)。[结论]快速型肉鸡肌纤维数量比慢速型肉鸡更多，IGF1 可以通过 PI3K/AKT 信号通路调控成肌纤维细胞的活力，从而促进肌纤维的发育。

Abstract

[Objectives] This paper aimed to elucidate the differences in myofiber structure and molecular regulatory mechanisms between fast- and slow-growing broilers, with a focus on characterizing myofiber developmental traits and their underlying molecular pathways. [Methods] Fast-growing Cobb500 broilers and slow-growing Wannan Sanhuang chickens were selected. Muscle samples were collected at embryonic 18 days (E18), 21 days post-hatch (D21), and 42 days post-hatch (D42). Myofiber morphology was analyzed using hematoxylin and eosin (HE) staining and scanning electron microscopy (SEM), while key myogenic gene expression was quantified via quantitative real-time PCR (RT-qPCR) and Western blotting. In vitro functional assays demonstrated that knockout and overexpression plasmids of insulin-like growth factor 1 (IGF1) were transfected into chicken embryo fibroblast (DF-1) cells to validate IGF1-mediated signaling pathways and biological functions. [Results] At embryonic 18 days (E18), the fast-growing broilers exhibited significantly smaller myofiber diameters compared to the slow-growing chickens (P<0.05). Conversely, myofiber diameters in the fast-growing broilers surpassed those of slow-growing chickens at 21- and 42-days post-hatch (P<0.05). Muscle fiber density in the fast-growing broilers was significantly higher at E18 (P<0.05) but markedly lower at D21 and D42 compared to the slow-growing chickens (P<0.05). IGF1 mRNA and protein expression levels in the fast-growing broilers were consistently elevated at E18, D21, and D42 relative to the slow-growing chickens (P<0.05). Overexpression of IGF1 significantly upregulated the expression of serine/threonine kinase 1 (AKT1), cyclin-dependent kinase 4 (CDK4), and c-Jun N-terminal kinase (JNK) genes compared to negative control and IGF1-knockdown groups (P<0.05). During 16-40 hours post-transfection, IGF1-overexpressing cells displayed significantly higher viability than IGF1-knockdown and blank control groups (P<0.05). [Conclusions] The fast-growing broilers possess a greater myofiber quantity than the slow-growing chickens. IGF1 enhanced myofiber development by activating the PI3K/AKT signaling pathway, thereby regulating myoblast viability and proliferation.

关键词

快速型肉鸡 / 慢速型肉鸡 / 肌纤维结构 / IGF1 / PI3K/AKT1 通路

Key words

fast-growing broilers / slow-growing broilers / myofiber structure / IGF1 / PI3K/AKT signaling pathway

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基金资助

安徽省重点研究和开发计划项目(202004a06020054)