抗阻运动对SAMP8小鼠骨骼肌lncRNA和mRNA基因表达谱的影响  

Effects of resistance exercise on long non-coding RNA andmRNA expression profiles of skeletal muscle in senescenceaccelerated-prone 8 mice

作　　者：傅泽铤 李仲豪 李伦宇 刘洪政 丁海丽[1] FU Zeting;LI Zhonghao;LI Lunyu;LIU Hongzheng;DING Haili(Institute of Sports Medicine and Health,Chengdu Sport University,Chengdu 610041,China;Guiyang Baiyun District Third Middle School,Guiyang 550014,China)

机构地区：[1]成都体育学院运动医学与健康研究所,成都610041 [2]贵阳市白云区第三中学,贵阳550014

出　　处：《中国实验动物学报》2024年第3期286-296,共11页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(81904318);四川省自然科学基金(2023NSFSC0545);四川省中央引导地方科技发展项目(2022ZYD0062);国家体育总局运动医学重点实验室(2018YFF0300904);运动医学四川省重点实验室(2021-A030);四川省科技创新创业苗子工程项目(2023JDRC0100)。

摘　　要：目的 利用RNA-seq技术比较运动组与对照组小鼠股四头肌中差异表达的lncRNA和mRNA,探讨抗阻运动作用于SAMP8小鼠的潜在调控机制。方法 12只28周龄雄性SAMP8快速衰老小鼠分为模型组(M组)、抗阻运动组(R组),每组6只;另设6只同龄SAMR1抗衰老小鼠作对照组(C组)。R组经递增负重爬梯运动训练8周。每1周测定相对抓力、每2周测转棒测试时间。取材后取右侧股四头肌进行苏木素-伊红(HE)染色观察其组织学变化;并取左侧股四头肌进行RNA-seq,筛选出差异表达的lncRNA(long non-coding RNA,lncRNA)和mRNA,然后对这些基因进行基因本体(Gene Ontology, GO)、京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)富集分析。结果 (1)干预前,SAMP8小鼠相比较于自然衰老的SAMR1小鼠表现出相对抓力以及加速转棒测试时长显著下降(P<0.01);干预后,与M组相比,R组相对抓力以及转棒时长均出现显著增加(P<0.01)。(2)HE染色结果显示,M组肌纤维横截面积与C组比较显著下降(P<0.01);R组肌纤维横截面积与M组比较显著增加(P<0.01)。(3)通过差异表达分析,相比于M组,R组有182个表达水平上调和218个表达下调的lncRNA以及454个表达上调和289个表达下调的mRNA。R组与M组之间差异lncRNA的靶基因KEGG显著富集的通路有产生IgA的肠道免疫网络、NF-κB、炎症性肠病等信号通路。结论 (1)本研究证明了抗阻运动能够改善SAMP8小鼠肌少症的骨骼肌机能,利用RNA-seq分别鉴定出M组、R组小鼠的差异lncRNA和mRNA。这些差异基因可能是肌少症的潜在治疗靶点。(2)通过分析差异表达lncRNA的靶基因以及mRNA的生物学信息,从而为进一步了解抗阻运动改善肌少症的机制提供了可能。Objective To explore the potential regulatory mechanism of resistance exercise in senescence accelerated-prone 8 mice(SAMP8)by evaluating the effects of exercise on the expression of long non-coding RNA(lncRNA)and mRNA in quadriceps muscles by RNA-sequencing(RNA-seq)technology.Methods Twenty-eight-week-old male SAMP8 mice were divided into a model group(M group)and resistance-exercise group(R group)(n=6 mice per group).Another eight normally aging SAMR1 mice of the same age were used as the control group(C group).Mice in R group received 8 weeks of increasing weight climbing exercise training.Relative grip strength was measured every week and the rotarod test was performed every 2 weeks.Histological changes in the right quadriceps femoris were observed by hematoxylin-eosin staining and the left quadriceps was used for RNA-seq.Differentially expressed lncRNA and mRNA were screened and analyzed for enrichment by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses.Finally,key differentially expressed genes were analyzed by quantitative reverse transcription-polymerase chain reaction to verify the accuracy of the RNA-seq result.Results(1)Relative grip strength and rotarod test time were significantly decreased in M group compared with C group(P<0.01),but were significantly increased after 8 weeks of Rgroup compared with M group(P<0.01).(2)The cross-sectional area of the muscle fibers was significantly lower in M group compared with C group,as shown by HE staining(P<0.01),while the cross-sectional area of the muscle fibers was significantly increased in the R group compared with M group(P<0.01).(3)Differential expression analysis identified 182 upregulated and 218 downregulated lncRNA,and 454 upregulated and 289 downregulated mRNA between M group and R group.The KEGG pathways of lncRNA target genes that were differentially expressed between M group and R group were significantly enriched in intestinal immune network for IgA production,nuclear factor-kappa B signaling pathway,and inflammatory bow

关 键 词：长非编码RNA SAMP8 肌少症 转录组 抗阻运动 

分 类 号：Q95-33[生物学—动物学]