ApoD对鸡成肌细胞增殖与分化的调控机制研究

Study on the Regulation Mechanism of ApoD on the Proliferation and Differentiation of Chicken Myoblasts

为揭示ApoD对鸡成肌细胞增殖与分化的调控作用,研究在鸡原代成肌细胞中分别过表达和干扰ApoD基因,检测胞内活性氧(ROS)和丙二醛(MDA)含量,同时通过流式细胞术、CCK-8、qRT-PCR及免疫印迹等试验探究其对细胞增殖和分化的影响。结果显示:ApoD降低胞内ROS和MDA含量,促进鸡原代成肌细胞增殖;同时,ApoD下调肌肉萎缩标志基因Atrogin1和MuRF1的表达,上调生肌因子MyoD、MyoG和MyHC的表达,从而促进鸡原代成肌细胞分化。研究结果提示ApoD通过降低胞内ROS含量、降低Atrogin1对MyoD的泛素化降解作用以及MuRF1对MyHC的抑制作用,促进鸡成肌细胞增殖与分化。

In order to reveal the regulatory effect of ApoD on proliferation and differentiation of chicken myoblasts, ApoD gene was overexpressed and interfered in chicken primary myoblasts in this study. Then the intracellular reactive oxygen species(ROS) content and malondialdehyde(MDA) content were detected and flow cytometry, CCK-8, qRT-PCR and western blot were performed to explore its effects on cell proliferation and differentiation. The results showed that ApoD reduced the intracellular ROS and MDA content and stimulated the proliferation of chicken primary myoblasts;At the same time, it down-regulated the expression of muscle atrophy marker genes Atrogin1 and MuRF1, and up-regulated the expression of myogenic factors MyoD, MyoG and MyHC, thus promoted the differentiation of chicken primary myoblasts. The results suggested that ApoD lowered the intracellular ROS content and reduced the ubiquitination and degradation of MyoD by Atrogin1 as well as the inhibition of MuRF1 on MyHC, thereby promoted the proliferation and differentiation of chicken myoblasts.