重庆医学2025，Vol.54Issue(12)：2759-2765,7.DOI:10.3969/j.issn.1671-8348.2025.12.008
小干扰RNA沉默甘氨酰-tRNA合成酶对L6细胞肌球蛋白重链Ⅱb表达的影响
Effect of glycyl-tRNA synthetase silencing by small interfering RNA on the expression of myosin heavy chain Ⅱb in L6 cells
摘要
Abstract
Objective To silencing glycyl-tRNA synthetase(GlyRS)in rat skeletal muscle myoblasts by using small interfering RNA(siRNA),and to investigate the effect GlyRS on the expression of myosin heavy chain Ⅱb(MHC-Ⅱb)in L6 cells.Methods Three GlyRS-siRNA sequences(GlyRS-siRNA1,2,and 3)were designed,synthesized,and transfected into L6 cells.The GlyRS mRNA expression was detected by qPCR to select the most efficient siRNA sequence.Another L6 cells were randomly divided into the normal cell group(no treatment,group N)and the negative control group(transfected with the negative control se-quence,group NC),the GlyRS-siRNA-24 h,-48 h,-72 h,-96 h and-120 groups(transfected with the GlyRS-siRNA sequence with the highest interference efficiency and cultured for the corresponding time),the average diameters of L6 cells in each group were measured at the corresponding time points respectively.the mRNA and pro-tein expressions of GlyRS and MHC-Ⅱb were detected by qPCR and Western blot respectively.Results Glyrs-siRNA1,2,and 3 decreased the expression level of GlyRS mRNA by 90.4%,62.0%,and 78.7%respectively.Among them,Glyrs-siRNA1 had the highest interference efficiency(P<0.001);the average diameter of L6 cells transfected with GlyRS siRNA1 for 24 h showed no significant change compared with the NC group.Af-ter transfection for 48 h and 72 h,the average diameter of L6 cells decreased significantly compared with the NC group(P<0.001),and then gradually increased.The expression level of GlyRS mRNA in each time point group of GlyRS-siRNA was lower than that in the NC group(P<0.05,P<0.01),among which the reduction was most obvious in the GlyRS-siRNA-72 h group.The expression levels of MHC-Ⅱb mRNA in each GlyRS-siRNA time point group were all higher than those in the NC group(P<0.05,P<0.01),among which the in-crease was more obvious in the GlyRS-siRNA-48 h,-72 h,and-96 h groups.But there was no statistically significant difference in pairwise comparisons among the three groups(P>0.05);The changes in the protein expression levels of GlyRS and MHC-Ⅱb were basically consistent with their mRNA expressions.Conclusion Silencing GlyRS can up-regulate the synthesis of MHC-Ⅱb in L6 cells.关键词
骨骼肌萎缩/甘氨酰-tRNA合成酶/小干扰RNA/肌球蛋白重链ⅡbKey words
skeletal muscle atrophy/glycyl-tRNA synthetase/small interfering RNA/myosin heavy chain Ⅱb分类
医药卫生引用本文复制引用
WANG Meng,LI Xucan,WANG Yu,YU Jiayan,YANG Lixia,WU Zhengxiao..小干扰RNA沉默甘氨酰-tRNA合成酶对L6细胞肌球蛋白重链Ⅱb表达的影响[J].重庆医学,2025,54(12):2759-2765,7.基金项目
贵州省科技计划项目(黔科合基础-ZK[2022]一般608) （黔科合基础-ZK[2022]一般608）
贵州省卫生健康委员会科学技术基金项目(gzwkj2021-505) （gzwkj2021-505）
遵义医学院硕士科研启动基金项目(F-891). （F-891）
