不同海拔牦牛骨骼肌中乳酸脱氢酶三种亚基基因表达的比较  

Comparison of Gene Expressions of Three Lactate Dehydrogenase Subunits in Skeletal Muscle of Yaks at Different Altitudes

作　　者：李琳 周娟 秦鸿楠 张勤文[1] LI Lin;ZHOU Juan;QIN Hong-Nan;ZHANG Qin-Wen(State Key Laboratory of Sanjiangyuan Ecology and Plateau Agriculture and Animal Husbandry,Department of Veterinary Medicine,College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,China)

机构地区：[1]青海大学农牧学院动物医学系,三江源生态与高原农牧业国家重点实验室,西宁810016

出　　处：《动物学杂志》2023年第2期277-284,共8页Chinese Journal of Zoology

基　　金：青海省自然科学基金项目(No.2023-ZJ-722)。

摘　　要：为了阐明高原低氧对牦牛(Bos mutus)骨骼肌中乳酸脱氢酶(LDH)三种亚基基因(LDHA、LDHB和LDHC)表达的影响,本实验分别选取高海拔(4 200 m)、中海拔(3 200 m)和低海拔(1 900 m)三个海拔位置养殖的临床健康成年雄性牦牛各5头,采用实时荧光定量PCR(qRT-PCR)和蛋白质印迹法检测牦牛骨骼肌中LDH三种亚基基因的m RNA表达和蛋白表达水平。结果表明,随海拔的升高,牦牛骨骼肌中LDHA m RNA的表达逐渐下降;LDHB m RNA先降低后升高,在高海拔组牦牛中表达最高,相对表达量为2.82±0.12,与低海拔组(1.01±0.07)、中海拔组(0.73±0.06)牦牛LDHB mRNA表达量差异显著(P <0.05);LDHC mRNA的表达量随海拔的升高呈下降趋势,且低海拔组(1.10±0.16)、中海拔组(0.86±0.16)、高海拔组(0.69±0.12)组间两两相比均差异显著(P <0.05)。LDHA和LDHC蛋白表达量随海拔的升高呈下降趋势,且LDHA蛋白表达量在低海拔组(1.00±0.00)、中海拔组(0.88±0.02)、高海拔组(0.75±0.02)组间两两相比均差异显著(P<0.05),LDHC蛋白表达量在低海拔组(1.00±0.00)、中海拔组(0.89±0.02)、高海拔组(0.74±0.02)组间两两相比差异显著(P <0.05);LDHB蛋白表达量随海拔的升高先降低再升高,高海拔组牦牛骨骼肌中LDHB蛋白的表达量(1.37±0.02)显著高于低海拔组(1.00±0.00)(P <0.05)和中海拔组牦牛(0.95±0.01)(P <0.05)。LDH是催化丙酮酸和乳酸转化的关键酶,其含量和活性受到基因的调控。本研究不同海拔间牦牛骨骼肌内LDH的m RNA表达和蛋白表达量均具有差异,推测高原牦牛骨骼肌为适应不同的氧分压环境,其丙酮酸和乳酸代谢发生了相应变化,无氧氧化代谢能力发生了适应性改变。[Objectives]In this study,we took Yak(Bos mutus)as studying object to illustrate the impact of low oxygen environment on the expression level of the three subunit genes(lactate dehydrogenase A,LDHA;lactate dehydrogenase B,LDHB;lactate dehydrogenase C,LDHC)of lactate dehydrogenase(LDH)gene in skeletal muscle.[Methods]Adult male Yaks that inhabit in different altitudes,including high altitude(altitude 4200 m),medium altitude(altitude 3200 m)and low altitude(altitude 1900 m)were used in our study.Real-time PCR and western blot were utilized to detect the mRNA and protein content of various skeletal muscle subtypes in Yaks at different altitudes.The experimental data were compared by one-way analysis of variance(ANOVA)using SPSS 23.0.[Results]The results showed that LDHA mRNA expression decreased in the Yak skeletal muscle with elevation increase(Fig.2).LDHB mRNA expression was first reduced and then increased in the Yak skeletal muscle with increasing elevation,and the expression of LDHB mRNA was highest among high-altitude Yaks(2.82±0.12),and the expression of LDHB mRNA at high altitude was significantly different from that of low-altitude(1.01±0.07)(P<0.05)and medium-altitude Yaks(0.73±0.06)(P<0.05)(Fig.2).The expression of LDHC mRNA decreased with elevation,and the difference between low altitude(1.10±0.16),medium altitude(0.86±0.16)and high altitude(0.69±0.12)was significant(P<0.05)(Fig.2).The expression of LDHA and LDHC proteins decreased with the increasing altitude,the expression of LDHA protein was significantly different between low altitude(1.00±0.00),medium altitude(0.88±0.02),and high altitude(0.75±0.02)(P<0.05)(Fig.3a,b),the expression of LDHC protein was significantly different between low altitude(1.00±0.00),middle altitude(0.89±0.02),and high altitude(0.74±0.02)(P<0.05)(Fig.3e,f);with an elevation in altitude,the expression of LDHB protein firstly reduced and subsequently returned to normal,and high-altitude Yaks(1.37±0.02)had significantly higher levels of LDHB protein expression in the

关 键 词：牦牛 乳酸脱氢酶 乳酸脱氢酶亚基 低氧适应 

分 类 号：S955[农业科学—水产养殖]