干扰DHX9对牛骨骼肌卫星细胞增殖与成肌分化的影响  

Effects of interfering DHX9 on the proliferation and myoblast differentiation of bovine skeletal muscle satellite cells

作　　者：杨旭 訾晶晶 于建杰 郭宏 丁向彬 刘新峰 张林林 YANG Xu;ZI Jingjing;YU Jianjie;GUO Hong;DING Xiangbin;LIU Xinfeng;ZHANG Linlin(College of Animal Science and Veterinary Medicine,Tianjin Agricultural University/Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry,Tianjin 300384,China;MOE Key Laboratory of Conservation and Utilization of Special Biological Resources in Western China,Ningxia University,Yinchuan 750021,China)

机构地区：[1]天津农学院动物科学与动物医学学院/天津市农业动物繁育与健康养殖重点实验室,天津300384 [2]宁夏大学西部特色生物资源保护与利用教育部重点实验室,宁夏银川750021

出　　处：《畜牧与兽医》2022年第2期9-14,共6页Animal Husbandry & Veterinary Medicine

基　　金：天津市自然科学基金(18JCYBJC95800);天津市教委科研计划项目(2018KJ183)。

摘　　要：为研究天冬氨酸-谷氨酸-丙氨酸-组氨酸盒解旋酶9(DEAH(Asp-Glu-Ala-His)-box helicase 9,DHX9)对牛骨骼肌细胞增殖与分化的影响,利用已经建立的牛骨骼肌卫星细胞体外成肌分化模型,设计合成DHX9的si-RNA,采用荧光定量PCR和Western blot技术检测DHX9基因在成肌分化前后以及增殖标志因子配对盒基因7(paired box 7, Pax7)、分化标志因子肌球蛋白重链(myosin heavy chain, MyHC)与肌细胞生成素(myogenin, MyoG)的mRNA与蛋白表达水平。结果显示:干扰后,DHX9在牛骨骼肌分化后表达量极显著下降(P<0.01);与对照组相比,增殖标志因子Pax7的mRNA水平和蛋白水平无显著变化(P>0.05),EdU阳性细胞率无变化(P>0.05),MyHC、MyoG的mRNA水平和蛋白水平均极显著升高(P<0.01);倒置显微镜观察发现肌管明显变粗。结果表明:干扰DHX9后对牛骨骼肌卫星细胞的增殖无影响,但能够促进牛骨骼肌卫星细胞的成肌分化过程,研究结果为进一步深入研究DHX9在动物骨骼肌发育分化调控机制方面提供参考。To determine the effects of DEAH(Asp-Glu-Ala-His)-box helicase 9(DHX9) on proliferation and differentiation of bovine skeletal muscle cells, the established model of in vitro myoblast differentiation of bovine skeletal muscle satellite cells was used, the si-RNA of DHX9 was designed and synthesized, and the mRNA expression level of DHX9 before and after the myoblast differentiation was detected by fluorescence quantitative PCR and Western blot. The MRNA and protein expression levels of proliferation marker Pax7, differentiation marker myosin heavy chain(MyHC) and myoietin(MyoG) were measured. The results showed that the expression of DHX9 decreased after the differentiation of the bovine skeletal muscles(P<0.01). There were no significant changes in the mRNA and protein levels of proliferation marker Pax7(P>0.05) and in the rate of EdU positive cells(P>0.05). The mRNA and protein levels of differentiation markers MyHC and MyoG were significantly increased(P<0.01). The present results indicated that interference with DHX9 had no effect on the proliferation of bovine skeletal muscle satellite cells, but it promoted the myogenic differentiation process of the bovine skeletal muscle satellite cells. The above finding provided a reference for further research on the regulation mechanism of DHX9 in animal skeletal muscle development and differentiation.

关 键 词：RNA/DNA解旋酶 牛骨骼肌 卫星细胞 成肌分化 

分 类 号：S823.9[农业科学—畜牧学]