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魏琳娜,魏莲,汪洋,李筱,许利娜,魏登邦.2015.高原鼠兔肝中Ldh-c基因的表达及其对无氧糖酵解水平的影响.动物学杂志,50(6):846-854.
高原鼠兔肝中Ldh-c基因的表达及其对无氧糖酵解水平的影响
The Function of the Sperm-specific Lactate Dehydrogenase Gene (Ldh-c) in Plateau Pika Liver
投稿时间:2015-01-07  修订日期:2015-09-28
DOI:DOI: 10.13859/j.cjz.201506004
中文关键词:  高原鼠兔  肝脏组织  Ldh-c基因  N-isopropyl oxamate  乳酸脱氢酶活力
英文关键词:Plateau pika (Ochotona curzoniae)  Liver tissue  Ldh-c gene  N-isopropyl oxamate  LDH enzymatic activity
基金项目:国家自然科学(Nos. 31260512);青海省自然科学(No.2012-Z-905);* 通讯作者,E-mail: weidengbang@163.com;第一作者介绍魏琳娜,女,硕士;研究方向野生动植物资源保护;E-mail:weilinna92@163.com。;国家自然科学基金项目(面上项目,重点项目,重大项目)
作者单位E-mail
魏琳娜 青海大学农牧学院 西宁810016 weilinna92@163.com 
魏莲 青海大学 西宁810016  
汪洋 青海大学医学院 西宁810016  
李筱 青海大学医学院 西宁810016  
许利娜 青海大学农牧学院 西宁810016  
魏登邦 青海大学 西宁810016 weidengbang@163.com 
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中文摘要:
      高原鼠兔(Ochotona curzoniac)对高原低氧环境有很强的适应性。我们研究发现,精子特异性乳酸脱氢酶(LDH-C4)在高原鼠兔肝脏组织中表达。为了为了阐明LDH-C4在高原鼠兔肝脏组织中的作用,本文应用荧光定量PCR和western blot方法,测定了精子特异性乳酸脱氢酶基因(Ldh-c)在高原鼠兔肝脏组织中的表达水平;应用对 LDH-C4 特异性的抑制剂(N-isopropyl oxamate),通过肌肉注射后,研究了抑制剂对高原鼠兔肝脏组织中LDH比活力,LD和ATP含量的影响。结果表明,在mRNA和蛋白水平,Ldh-c基因在高原鼠兔肝脏组织中均有表达,相对表达水平分别为0.22±0.04和0.97 ±0.20;当肌肉注射1ml 1M的抑制剂时,血液中抑制剂浓度为0.08mM,肝脏组织中LDH比活力(抑制剂组和空白对照组LDH比活力分别为4.99±0.29U/mg?prot和7.36±0.42U/mg?prot)、LD(抑制剂组和空白对照组LD含量分别为0.38±0.05 mmol/g?prot和0.53±0.03mmol/g?prot)和ATP(抑制剂组和空白对照组ATP含量分别为5.84±0.83nmol/mg?prot和7.78±1.06nmol/mg?prot)含量显著下降,抑制剂对LDH,LD和ATP的抑制率分别为30.19%、32.22%和24.94%。以上结果说明,Ldh-c在高原鼠兔肝脏组织中表达,LDH-C4通过催化无氧糖酵解过程,为其肝脏组织生命活动提供至少24%的ATP,这使高原鼠兔减小了在低氧环境中对氧气的依赖,增强了对低氧环境的适应力。
英文摘要:
      The plateau pika (Ochotona curzoniae) has a strong adaptability to hypoxic plateau environment. We found that the sperm-specific lactate dehydrogenase (LDH-C4) gene expressed in plateau pika liver. In order to shed light on the function of LDH-C4 in plateau pika liver, we determined the expression levels of Ldh-c gene in plateau pika liver by real-time PCR and western blot. After injecting N-isopropylSoxamate, a LDH-C4 specific inhibitor, in skeletal muscle of hind legs, the activities of LDH, and the contents of LD and ATP in plateau pika liver were measured. The results indicated that the expression levels of Ldh-c mRNA and protein were 0.22±0.04 and 0.97±0.20, respectively. After 30 minutes injectedS1 ml 1 M N-isopropylSoxamate in biceps femorisSmuscle, the concentration of N-isopropyl oxamate in blood was 0.08 mM. In the liver of inhibitor group and control group, the LDH activities were 4.99±0.29 U/mg?prot and 7.36±0.42 U/mg?prot, the contents of LD were 0.38±0.05 mmol/g?prot and 0.53±0.03 mmol/g?prot, and ATP were 5.84±0.83 nmol/mg?prot and 7.78±1.06 nmol/mg?prot, respectively. The LDH activities, the contents of LD and ATP in liver of the inhibitor group were decreased significantly, and the inhibition rate of N-isopropyl oxamateSto LDH, LD and ATPSin plateau pika liver were 30.19%、32.22% and 24.94%, respectively. These results indicated that the sperm-specific lactate dehydrogenase gene expressed in plateau pika liver, the pika liver got at least 24% ATP for its life activities by catalyzing anaerobic glycolysis. As a result, the plateau pika reduced dependence on oxygen and enhanced the adaptation to the hypoxic environments.
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