基于RNA测序技术研究推拿改善运动性疲劳大鼠肝损伤的作用机制  

Study on the mechanism of action of Tuina to improveliver injury in exercise-induced fatigue rats based on RNA sequencing technology

作　　者：宋红志 王泽中 钮妍 付国兵[3] 魏培栋[3] 蔡明亨 王康[3] SONG Hongzhi;WANG Zezhong;NIU Yan;FU Guobing;WEI Peidong;CAI Mingheng;WANG Kang(Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China)

机构地区：[1]北京中医药大学第二临床医学院,100078 [2]北京中医药大学第三附属医院儿科 [3]北京中医药大学东方医院推拿理疗科

出　　处：《环球中医药》2024年第5期794-800,共7页Global Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81574092)。

摘　　要：目的应用RNA测序技术探讨推拿改善运动性疲劳大鼠肝损伤的作用机制。方法6周龄雄性SD大鼠,随机分入对照组、模型组和推拿组,每组8只。对照组常规喂养,不做任何处理;模型组每日力竭运动后于笼中休息;推拿组每日力竭运动后予推拿干预15分钟,各组共干预21天。取材后,运用组织病理学方法观察各组大鼠的肝脏组织形态学变化以及TUNEL检测观察大鼠肝脏细胞凋亡情况;提取各组大鼠肝脏组织RNA进行转录组测序,并对差异表达基因进行GO富集分析和KEGG通路分析,确定其生物学功能。结果与对照组相比,两组大鼠均出现了轻度肝损伤的病理改变,且模型组大鼠的肝脏细胞凋亡率较对照组显著上升(P<0.05);与模型组相比,推拿组大鼠肝脏组织病理改变均明显改善,肝窦微狭窄,小叶间有少量炎症细胞浸润,肝脏细胞趋于正常,且推拿组大鼠肝脏细胞凋亡率较模型组显著下降(P<0.01)。经转录组学分析,对照组与模型组共筛选出差异基因491个,其中上调基因244个,下调基因247个,模型组和推拿组共筛选出差异基因513个,其中上调基因257个,下调基因256个,两组共有的差异显著基因筛选出5个,分别为Ppargc1a、Fbp2、Gadd45g、Atp1b2和Myh7。GO富集分析结果显示:推拿改善运动性疲劳大鼠肝脏损伤主要涉及信号传递、对刺激的反应以及核酸结合转录因子活性等;KEGG通路分析结果主要包括FoxO信号通路、AMPK信号通路和cGMP-PKG信号通路等。结论推拿改善了运动性疲劳引起的大鼠肝脏组织的炎性损伤及细胞凋亡,其作用机制可能与Ppargc1a、Fbp2、Gadd45g、Atp1b2和Myh7基因高度相关。Objective Application of RNA sequencing technology to explore the mechanism of action of Tuina to improve liver injury in exercise-induced fatigue rats.Methods Six-week-old male Sprague-Dawley rats were randomly assigned into the control group,the model group,and the Tuina group,with 8 rats in each group.The control group was fed routinely without any treatment;rats in the model group was rested in a cage after daily exhaustive exercise;and ats in the Tuina group was given Tuina for 15 min after daily exhaustive exercise for a total of 21 days.After sampling,the histopathological methods of light microscopy were used to observe the morphological changes of liver tissues of the three groups of rats,and TUNEL assay was used to observe the apoptosis of rat liver cells;RNA of liver tissues of the three groups of rats was extracted for transcriptome sequencing,and differentially expressed genes were screened and subjected to GO enrichment analysis and KEGG pathway analysis to determine their biological functions.Results Compared with the control group,both groups of rats showed pathological changes of mild liver injury,and the rate of apoptosis in the liver cells of rats in the model group was significantly increased compared with that of the control group(P<0.05).Compared with the model group,the compared with the model group,the pathological changes in the liver tissue of the Tuina group rats were significantly improved,with slight narrowing of the liver sinusoids and a small amount of inflammatory cell infiltration between lobules.The liver cells tended to be normal,and the rate of apoptosis in the liver of rats in the Tuina group was significantly decreased compared with that in the model group(P<0.01).After transcriptomic analysis,491 differential genes were screened in the control group and the model group,including 244 up-regulated genes and 247 down-regulated genes.513 differential genes were screened in the model group and the Tuina group,including 257 up-regulated genes and 256 down-regulated genes.There wer

关 键 词：推拿 运动性疲劳 转录组学 肝脏损伤 细胞凋亡 信号通路 

分 类 号：R244.1[医药卫生—针灸推拿学]