| 李琳,周娟,秦鸿楠,张勤文.2023.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.动物学杂志,58(2):277-284. |
| 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 |
| 投稿时间：2022-08-20 修订日期：2023-03-14 |
| DOI：10.13859/j.cjz.202302011 |
| 中文关键词: 牦牛 乳酸脱氢酶 乳酸脱氢酶亚基 低氧适应 |
| 英文关键词:Yak Lactate dehydrogenase Lactate dehydrogenase subunit Hypoxia adaptation |
| 基金项目:青海省自然科学基金项目（No. 2023-ZJ-722） |
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| 中文摘要: |
| 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mRNA 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 mRNA 的表达逐渐下降；LDHB mRNA 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mRNA 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。 |
| 英文摘要: |
| [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
4 200 m), medium altitude (altitude 3 200 m) and low altitude (altitude 1 900 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 their skeletal muscles than did low-altitude (1.00 ± 0.00) (P <
0.05) and medium-altitude Yaks (0.95 ± 0.01) (P < 0.05) (Fig. 3c, d). [Conclusion] Lactate dehydrogenase is
the key enzyme catalyzing conversion of pyruvate and lactate, its content and activity are regulated by genes.
The results of this experiment report different mRNA expression and protein content of LDH in Yak skeletal
muscle at different altitudes, which indicate adaptation of skeletal muscle in plateau-living Yaks to different
oxygen partial pressure environments, as well as corresponding changes in pyruvate and lactic acid
metabolism in skeletal muscle, and the oxidative metabolism capacity of anaerobic oxidation. |
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