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龚丽景,贾杰,付鹏宇.基于 RNA 测序分析不同低氧暴露模式对大鼠胫骨前肌差异基因表达的影响[J].中国实验动物学报,2022,30(1):84~91.
基于 RNA 测序分析不同低氧暴露模式对大鼠胫骨前肌差异基因表达的影响
RNA sequencing analysis of differentially expressed genes of the anterior tibial muscle in rats after different modes of hypoxia exposure
投稿时间:2021-08-12  
DOI:10. 3969 / j.issn.1005-4847. 2022. 01. 011
中文关键词:  慢性间歇低氧  急性低氧  骨骼肌  RNA 测序
英文关键词:chronic intermittent hypoxia  acute hypoxia  skeletal muscle  RNA sequencing
基金项目:
作者单位E-mail
龚丽景 北京体育大学 运动与体质健康教育部重点实验室,北京 100084 lijing.gong@ bsu.edu.cn 
贾杰 北京体育大学 运动人体科学学院, 北京 100084  
付鹏宇 西北工业大学体育部,西安 710072 1402884452@ qq.com 
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中文摘要:
       目的 筛选慢性间歇低氧暴露和急性低氧暴露对大鼠胫骨前肌差异表达基因及其相关通路分析。 方法 SD 大鼠 24 只,分为常氧对照组(C 组)、慢性间歇低氧组(IH 组,氧浓度为 12. 4%,每天 8 h,共 4 周)和急性低氧组(AH 组,氧浓度为 12. 4%,每天 24 h,共 3 d)。 干预后,测试抓力和瘦体重,取胫骨前肌(TA)进行 HE 染色后,再统计肌纤维横截面积(FCSA),Western Blot 法测试 Atrogin-1 和 MuRF1 的蛋白相对含量,并提取大鼠 TA 总 RNA 进行测序,筛选其差异基因,并分析相关的生物过程(BP)和通路(pathway)。 结果 (1)干预期间各时间点 IH 组体重低于 C 组,AH 组体重持续下降;干预后,AH 组瘦体重和相对抓力显著低于 C 组(P< 0. 05)。 (2)IH 组和 AH 组肌纤维形态出现损伤,且 IH 组更加明显,而各组 TA 的 FCSA 无显著差异。 (3)IH 组 TA 中 Atrogin-1 和 MuRF1 含量显著均高于 C 组,AH 组中 MuRF1 含量显著高于 C 组(P< 0. 05)。 (4)筛选差异基因发现,在 AH/ C 组上调而在 IH/ C 组下调的差异基因为慢肌结构相关基因。 GO 和 KEGG 分析可知,IH/ C 组差异基因主要富集于 PPAR 通路;上调差异基因功能富 集在抗氧化和糖脂代谢过程,下调基因富集在快慢肌之间转化和氧化应激过程;AH/ C 组上调差异基因功能主要富集于氧化应激、炎症反应和快肌和慢肌之间的转化,下调基因富集于 smad 蛋白信号转导和泛素蛋白连接酶结合等过程。 结论 慢性间歇低氧和急性低氧影响大鼠胫骨前肌差异表达分别富集于影响糖脂代谢水平和促进氧化应激及炎症反应上,说明不同低氧模式可通过不同转录途径而影响骨骼肌的代谢。
英文摘要:
       Objective Screening for differentially expressed genes (DEGs) in anterior tibial muscle of rats and pathway analysis after chronic intermittent hypoxia and acute hypoxia exposure. Methods Twenty-four SD rats were randomly divided into the normoxia control group (C group), the chronic intermittent hypoxia group ( IH group, 12. 4% O2 , 8 h / d, 4 weeks) and the acute hypoxia group (AH group, 12. 4% O2 , 24 h / d, 3 d). The grasping force and lean body weight were examined after the interventions. The anterior tibial muscle (TA) fiber cross-sectional area (FCSA) was examined by hematoxylin / eosin staining. Relative protein expression levels of Atrogin-1 and muscle RING-finger protein-1 (MuRF1) were examined by western blotting. Total RNA of TA was extracted and sequenced, DEGs were screened, and the biological process (BP) and pathways enriched by DEGs were analyzed. Results (1) During the intervention, body weights of the IH group were lower than those of the C group at each time point, and the body weight of AH group continued to decrease. After the intervention, the lean body weight and relative grip in the AH group were significantly lower than in the C group (P< 0. 05). (2) The morphology of muscle fibers of the TA was destroyed in the IH and AH groups, and was more obvious in the IH group, but there were no significant differences in FCSA. (3) The levels of Atrogin-1 and MuRF1 in TA in the IH group were significantly higher than those in the C group, and the MuRF1 level in the AH group was significantly higher than the C group ( P< 0. 05). ( 4) The DEGs up-regulated in AH/ C group comparison but down- regulated in the IH/ C group comparison were slow muscle structure-related genes. Gene Ontogeny and KEGG analysis showed that the DEGs in the IH/ C group were mainly enriched in peroxisome proliferator-activated receptor signaling pathway and other pathways. The function of up-regulated DEGs was mainly concentrated in the process of antioxidant and glycolipid metabolism, and for down-regulated genes were concentrated in the processes of transformation between fast and slow muscles and oxidative stress. The functions of up-regulated DEGs in the AH/ C group were mainly concentrated in oxidative stress, inflammatory response and transformation between fast and slow muscles, and for the down-regulated DEGs were concentrated in Smad protein signal transduction and ubiquitin protein ligase binding. Conclusions The DEGs of AT of rats affected by chronic intermittent hypoxia and acute hypoxia were enriched for processes in glycolipid metabolism and promoting oxidative stress and inflammatory responses, respectively, suggesting that different hypoxia modes affect skeletal muscle metabolism through different pathways.
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