摘要
目的:通过基因芯片等方法寻找血流限制诱导肌肥大组织表达变化的基因,探讨其原因和分子机制。方法:2019年9月~2019年11月,选用SPF级健康SD大鼠8只,适应性喂养1周后,称重,采用自身左右后肢对照。所有大鼠按剂量30mg/kg体重戊巴比妥钠麻醉后在右后肢膝盖上方近股关节处橡皮带束缚血流限制,计时30min,血流限制每周2次,共4次。动物处理后正常饲养和活动。造模完成后取出双侧下肢比目鱼肌作为样本,用DEPC水漂洗后投入液氮中保存,抽提大鼠肌肉组织mRNA,经反转录获取大鼠比目鱼肌组织cDNA探针,进行基因芯片检测。检测结果经荧光定量聚合酶链反应(PCR)验证。结果:与对照组大鼠的比目鱼肌组织相比,实验组比目鱼肌组织的8条差异表达基因,其中6条显著上调基因(Chrna1、Cacnb1、Pla2g2a、Stap2、Hspbap1和Trib3)和2条显著下降基因(Ipo5和Smc4)。在8条基因中6条(75%)为已知基因,已在GeneBank中登录;2条(25%)基因为未知基因。已知基因中6条全部表达上调,下调表达基因全部为未知基因。RT-PCR验证结果与基因芯片检测结果相符。结论:血流限制诱导骨骼肌肥大的发生发展是多种因素、多条途径相互作用的结果。实验结果提示所有差异表达基因中的上升和下降基因,主要涉及肌纤维蛋白合成与收缩相关介质和通道有关;差异表达基因中还有很多未知基因,许多相关的未知领域有待研究。
Objectives:To identify significantly differentially expression genes of muscle hypertrophy in rats at skeletal muscle blood restriction,and to find out the reasons and molecular mechanisms that cause muscle hypertrophy with blood restriction.Methods:SPF level 8 healthy SD rats were selected with its left and right hind limb control.All rats at dose of 30 mg/kg in anesthesia,were restricted in blood flow with the right hind leg above the knee joints,30 minutes a time with total four times,two times a week.After the success of modeling,remove the bilateral soleus muscle as a sample which was preserved in liquid nitrogen after rinsing by DEPC water.Muscle tissue of rats with mRNA extraction,reverse transcription cDNA probe by obtaining the soleus muscle tissue of rats.The test results were verified fluorescence quantitative polymerase chain reaction(PCR).Results:Compared with the control group of rats soleus muscle tissue.There is a significant change in 8 gene expression in soleus muscle tissue of the experimental group,which had 6 genes up regulated and 2 genes significantly down regulated expression.The all known genes are up-regulated and down regulated genes are all unknown gene.The RT-PCR verification results are consistent with the results measured by gene chip.Conclusions:The skeletal muscle hypertrophy after blood restriction is the result of the interaction of multi way and variety of factors,the experimental results suggested that all the differentially expressed genes in the rising and falling,mainly related to protein synthesis and muscle fiber contraction associated media and channels.A lot of unknown genes are to be studied,there are still many unknown fields that muscle hypertrophy after blood restriction.
作者
李高权
徐守宇
刘雪云
王珊玺
LI Gao-quan;XU Shou-yu;LIU Xue-yun;WANG Shan-xi(Department of rehabilitation medicine in XiangNan University,Chenzhou,Hunan 423000;Department of Orthopaedics,Juntendo University School of Medicine,Tokyo,Japan 113-8421;Department of Rehabilitation,Third Clinical Medical College,Zhejiang Chinese Medical University,Hangzhou,Zhejiang 310053)
出处
《按摩与康复医学》
2021年第5期64-67,共4页
Chinese Manipulation and Rehabilitation Medicine
基金
湖南省教育厅科学研究项目,编号:16C1486
湘南学院应用特色学科,编号:校发[2020]58号。
关键词
骨骼肌
肌肥大
大鼠
血流限制
基因组学
skeletal muscle
muscle hypertrophy
rat
blood restriction
genomics
