摘要
为探究糖酵解途径关键基因干扰对其他肉品质功能基因的影响,先采用脂质体转染技术向分离、培养的肌肉卫星细胞中转染siRNA,干扰4个糖酵解途径关键基因(PKM、PFKM、PGK1、ALDOA)的表达,再利用经诱导分化的肌肉卫星细胞进行RNA提取和反转录,并使用qPCR方法检测4个目的基因及15个肉品质功能基因的mRNA表达情况。结果表明,与对照组相比,转染组细胞在增殖特点和表型上没有明显差异,转染组4个目的基因的表达量下降33.5%~93.8%。PKM干扰后,7个肉品质功能基因表达显著下调,5个显著上调;PFKM和PGK1干扰后,12个肉品质功能基因表达显著下调,3个显著上调;ALDOA干扰后,8个肉品质功能基因表达显著下调,3个上调。在4个转染组中,LDHA、ACOX1、PNPLA、CAPN1基因的表达水平均发生显著下降,而I-FABP、POMC、DGAT1基因的表达水平上调最明显,相关基因主要涉及细胞能量代谢、糖脂代谢以及细胞组织结构形成通路,其中脂质代谢和能量调节通路基因在目的基因干扰下受到的影响最大。研究结果为在基因水平探究肉品质形成的内在调控机制提供了参考。
To explore the interference impacts of 4 glycolysis pathway genes on other meat quality related genes,liposome transfection technology was used to transfect siRNAs of 4 glycolysis pathway genes(PKM,PFKM,PGK1,ALDOA)into the isolated and cultured satellite cells of goat muscle.The mRNA expressions of 4 target genes and 15 meat quality related genes were detected by qPCR after induced differentiation,RNA extraction and reverse transcription.The results showed that there was no significant difference in cell proliferation characteristics and phenotypes between the control and the transfection groups,and the expression levels of 4 target genes decreased 33.5%to 93.8%after interference.The expressions of 7 meat quality related genes were significantly down-regulated and 5 genes were up-regulated after PKM interference,the expressions of 12 meat quality related genes were significantly down-regulated and 3 genes were up-regulated under the interference of PFKM and PGK1,and the expressions of 8 meat quality related genes were significantly down-regulated and 3 genes were up-regulated after ALDOA interference.The expression levels of LDHA,ACOX1,PNPLA,CAPN1genes in the four transfection groups decreased significantly,while the expression levels of I-FABP,POMC,DGAT1genes increased obviously.The above changed genes mainly involved in cellular energy metabolism,glucose and lipid metabolism,cell tissue structure formation pathways,among which lipid metabolism and energy regulation genes suffered most under the interference of the target genes.These results provide a reference for exploring the internal regulatory mechanism of meat quality formation at gene level.
作者
刘永峰
李衡
付尚辰
肖宇
LIU Yongfeng;LI Heng;FU Shangchen;XIAO Yu(School of Food Engineering and Nutritional Science,Shaanxi Normal University,Xi an 710119,Shaanxi,China)
出处
《陕西师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2022年第5期90-98,共9页
Journal of Shaanxi Normal University：Natural Science Edition
基金
国家自然科学基金(U1903109)
陕西省重点研发计划(2021NY-032,2022NY-028)
西安市科技计划(20NYYF0011)。
