PDK1、HIF-1α 及 VEGF 在不同年龄牦牛肺脏中的表达差异

doi:10.3969/j.issn.1674-7968.2023.08.006

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摘要丙酮酸脱氢酶激酶 1 (pyruvate dehydrogenase kinase 1, PDK1) 低氧下可由缺氧诱导因子 1α (hypoxia inducible factor 1α, HIF-1α)诱导并参与调控糖酵解反应,同时可上调 HIF-1α的靶基因血管内皮生长因子(vascular endothelial growth factor, VEGF)表达,促进肺组织结构的重塑。为了揭示 PDK1、HIF-1α 及 VEGF 因子在不同年龄牦牛(Bos grunniens)肺脏中的表达分布特点,探讨其在牦牛肺脏适应高原低氧环境可能的调控作用,本研究通过采集新生、6 月龄、3 岁、6 岁健康牦牛的肺脏组织,运用苏木精-伊红 (hematoxylin-eosin, HE)染色、免疫组织化学染色、光密度分析、qRT-PCR 和蛋白免疫印迹的方法,对不同年龄组牦牛肺脏组织中PDK1、HIF-1α 及 VEGF的分布以及表达量的差异性进行研究。HE 染色结果显示,随着年龄的增长,终末细支气管管壁平滑肌增厚。免疫组化和光密度分析结果显示,PDK1、HIF-1α 及 VEGF 主要分布在不同年龄牦牛肺终末细支气管及其分支的单层纤毛柱状上皮以及伴行动脉、肺泡壁上。qRT-PCR 结果显示,PDK1 和HIF-1α 基因的相对表达量在6 岁组表达量最高,3 岁组表达次之。VEGF 基因的相对表达量在4 个年龄组均差异性不显著。Western blot 结果显示,PDK1 蛋白在6 岁组的表达量显著低于新生、6 月龄和3 岁组(P<0.05),HIF-1α 和VEGF 蛋白在6 岁组的表达量显著高于新生、6 月龄和3 岁组(P<0.05)。PDK1的蛋白表达量会随着年龄的增长先上升后下降,HIF-1α 和VEGF 则呈现升高的趋势,推测 PDK1、HIF-1α 及 VEGF 蛋白在低氧下可调节肺内葡萄糖代谢和血管重塑过程。综上,PDK1、 HIF-1α 及 VEGF 可能在牦牛肺脏发育过程中起重要作用,并参与调控牦牛适应高原低氧环境。本研究为不同年龄牦牛高原低氧适应性机制的比较研究提供了基础资料。

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	张伊阳
	陈树吾
	李斯佳
	李睿
	周熳琳
	杨琨
	张兰
	姚一凡
	张倩
	乔自林

关键词 ：牦牛, 肺脏, 丙酮酸脱氢酶激酶 1 (PDK1), 缺氧诱导因子 1α, (HIF-1α, ), 血管内皮生长因子(VEGF)

Abstract：Pyruvate dehydrogenase kinase 1 (PDK1) could be induced by hypoxia inducible factor 1α (HIF-1α) and participated in the regulation of glycolysis. The expression of vascular endothelial growth factor (VEGF) which is the target gene of HIF-1α up-regulated by PDK1. Furthermore, the PDK1 could promote the remodeling of lung structure. To reveal the expression and distribution characteristics of PDK1, HIF-1α and VEGF in the lungs of yaks (Bos grunniens) at different ages, and to explore their possible regulatory role in the adaptation of yak lungs to the hypoxia environment of the plateau, in this study, the lung tissues of healthy yaks aged newborn, 6-month-old, 3-year-old, and 6-year-old were collected. Hematoxylin-eosin (HE) staining, immunohistochemical staining, optical density analysis, qRT-PCR and Western blot were used to investigate the distribution and expression of PDK1, HIF-1α and VEGF in lung tissues of yaks in different age groups. The results of HE staining showed that the smooth muscle of the terminal bronchiole wall was thickened with age. The immunohistochemical and densitometric analysis results showed that PDK1, HIF-1α and VEGF were mainly distributed in the single-layer ciliated columnar epithelium of terminal bronchioles and their branches, as well as in the associated arteries and alveolar walls of yaks in different ages. Moreover, qRT-PCR results showed that the relative expressions levels of PDK1 and HIF-1α were the highest in the 6-year-old group, followed by the 3-year-old group. And there was no significant difference in the relative expression of VEGF among the 4 age groups. Additionally, the Western blot results showed that the expression of PDK1 protein in the 6-year-old group was significantly lower than that in newborn, 6-month-old and 3-year-old groups. The expression of HIF-1α and VEGF protein in the 6-year-old group was significantly higher than that in the newborn, 6-month-old and 3-year-old groups. Consequently, the protein expression of PDK1, HIF-1α and VEGF were negatively correlated with age. Expression of PDK1 increased and then decreased with age, while HIF-1α and VEGF showed an increasing trend. It was speculated that PDK1, HIF-1α and VEGF proteins could regulate glucose metabolism and vascular remodeling processes in the lung under hypoxia. In conclusion, PDK1, HIF-1α and VEGF might play an important role in the development of yaks lung and participate in the regulation of yaks adaptation to plateau hypoxia environment. This study provides basic data for comparative study of the adaptation mechanism in yaks of different ages to high altitude hypoxia.

Key words： Yak Lung Pyruvate dehydrogenase kinase 1 (PDK1) Hypoxia inducible factor 1α (HIF-1α) Vascular endothelial growth factor (VEGF)

收稿日期: 2022-05-11

ZTFLH:

S823.8+5

基金资助:国家自然科学基金 (31860687); 甘肃省自然科学基金 (21JR11RA024); 中央高校基本科研业务费项目 (31920230060); 甘肃省教育科技创新项目(2023B-055); 教育部动物医学生物工程创新团队(IRT-17R88); 国家自然科学基金 (32002241)

通讯作者: * 186152592@xbmu.edu.cn

引用本文:

张伊阳, 陈树吾, 李斯佳, 李睿, 周熳琳, 杨琨, 张兰, 姚一凡, 张倩, 乔自林. PDK1、HIF-1α 及 VEGF 在不同年龄牦牛肺脏中的表达差异[J]. 农业生物技术学报, 2023, 31(8): 1623-1631.
ZHANG Yi-Yang, CHEN Shu-Wu, LI Si-Jia, LI Rui, ZHOU Man-Lin, YANG Kun, ZHANG Lan, YAO Yi-Fan, ZHANG Qian, QIAO Zi-Lin. Differential Expression of PDK1, HIF-1α and VEGF in the Lung of Yak (Bos grunniens) at Different Age. 农业生物技术学报, 2023, 31(8): 1623-1631.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2023.08.006 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2023/V31/I8/1623

[1] 何俊峰, 余四九, 崔燕. 2009. 不同年龄高原牦牛肺脏的组织结构特征[J]. 畜牧兽医学报, 40(5): 748-755.
(He J F, Yu S J, Cui Y.2009. Characteristics of lung structure in different age plateau yak[J]. Acta Veterinaria et Zootech-nica Sinica, 40(5): 748-755.)
[2] 李小静, 刘川川, 刘辉琦, 等. 2017. 低氧对大鼠血清 bFGF、 VEGF 的表达和肺动脉超微结构的影响[J]. 中国高原医学与生物学杂志, 38(03): 164-170.
(Li X J, Liu C C, Liu H Q, et al. 2017. Effect of hypoxia on the expres-sion of bFGF, VEGF and the ultrastructural changes of pulmonary artery in rats[J]. Chinese High Altitude Medi-cine and Biology, 38(03): 164-170.)
[3] 闫明宇, 杨敬平, 赵永祥. 2017. 低氧性肺动脉高压大鼠肺组织中bFGFmRNA 及 VEGFmRNA 的表达与肺血管重构的关系[J]. 中国冶金工业医学杂志, 34(04): 373-375.
(Yan M Y, Yan J P, Zhao Y X.2017. The relation-ship of the expression of bFGFmRNA and VEGFmRNA withpulmonary vascular remodeling in rats lung tissue with hypoxic pulmonary hypertension[J]. Chinese Medi-cal Journal of Metallurgical Industry, 34(04): 373-375.)
[4] Boussat S, Eddahibi S, Andre C, et al. 2000. Expression and regulation of vascular endothelial growth factor in hu-man pulmonary epithelial cells[J]. American Journal of Physiology-Lung Cellular & Molecular Physiology, 279(2): 371-8.
[5] Di R, Yang Z, Xu P, et al. 2019. Silencing PDK1 limits hypox-iainduced pulmonary arterial hypertension in mice via the Akt/p70S6K signaling pathway[J]. Experimental and Therapeutic Medicine, 18(1): 699-704.
[6] He Y, Munday J S, Perrott M, et al. 2019. Association of age with the expression of hypoxia-inducible factors HIF-1α, HIF-2α, HIF-3α and VEGF in lung and heart of ti-betan sheep[J]. Animals, 9(9): 673.
[7] Honda T, Hirakawa Y, Nangaku M.2019. The role of oxida-tive stress and hypoxia in renal disease[J]. Kidney Re-search and Clinical Practice, 38(4): 414-426.
[8] Hui R, Birch N P, Suresh V.2016. An optimised human cell culture model for alveolar epithelial transport[J]. PLOS ONE, 11(10): 1-22.
[9] Jackson S G, Zhang Y, Haslam R J, et al. 2007. Structural analysis of the carboxy terminal PH domain of pleck-strin bound to D-myo-inositol 1, 2, 3, 5, 6-pentakisphos-phate[J]. BMC Structural Biology, 7(1): 1-11.
[10] Lan D, Xiong X, Ji W, et al. 2018. Transcriptome profile and unique genetic evolution of positively selected genes in yak lungs[J]. Genetica, 146(2):151-160.
[11] Liu J, Wang W, Wang L, et al. 2018. IL-33 initiates vascular remodelling in hypoxic pulmonary hypertension by up- regulating HIF-1α and VEGF expression in vascular en-dothelial cells[J]. Ebiomedicine, 33: 196-210.
[12] Liu L X, Lu H, Luo Y, Date T, et al. 2002. Stabilization of vas-cular endothelial growth factor mRNA by hypoxia-in-ducible factor 1[J]. Biochemical and Biophysical Re-search Communications, 291(4): 908-914.
[13] Ma X, Li C, Sun L, et al. 2014. Lin28/let-7 axis regulates aer-obic glycolysis and cancer progression via PDK1[J]. Na-ture Communications, 5(1): 1-13.
[14] Muhammad S A, Norika C, Chang H S, et al. 2022. HIF -1α plays an essential role in BMP9-mediated osteoblast dif-ferentiation through the induction of a glycolytic en-zyme, PDK1[J]. Journal of Cellular Physiology, 237(4):2183-2197.
[15] Park H, Lee S, Shrestha P, et al. 2015. AMIGO2, a novel membrane anchor of PDK1, controls cell survival and angiogenesis via Akt activation[J]. Journal of Cell Biolo-gy, 211(3): 619-637.
[16] Pearl L, Navdeep S C, Simon M C.2020. Cellular adaptation to hypoxia through hypoxia inducible factors and be-yond[J]. Nature Reviews Molecular Cell Biology, 21(5):268-283.
[17] Primo L, Di B L, Roca C, et al. 2007. Essential role of PDK1 in regulating endothelial cell migration[J]. Journal of Cell Biology, 176(7): 1035-1047.
[18] Qin L, Tian Y, Yu Z, et al. 2016. Targeting PDK1 with dichlo-roacetophenone to inhibit acute myeloid leukemia (AML) cell growth[J]. Oncotarget, 7(2): 1395-1407.
[19] Semba H, Takeda N, Isagawa T, et al. 2016. HIF-1α-PDK1 ax-is-induced active glycolysis plays an essential role in macrophage migratory capacity[J]. Nature Communications, 7: 11635-11644.
[20] Seong H A, Jung H, Choi H S, et al. 2005. Regulation of trans-forming growth factor- β signaling and PDK1 kinase ac-tivity by physical interaction between PDK1 and serine- threonine kinase receptor-associated protein[J]. Journal of Biological Chemistry, 280(52): 42897-42908.
[21] Tucker A, Mcmurtry I, Reeves J, et al. 1975. Lung vascular smooth muscle as a determinant of pulmonary hyperten-sion at high altitude[J]. American Journal of Physiology,228(3): 762-767.
[22] Yang Y, Kong W, Xia Z, Xiao L, et al. 2016. Regulation mech-anism of PDK1 on macrophage metabolism and function[J]. Cell Biochemistry and Function, 34(8): 546-553.
[23] Zhang H, He H, Cui Y, et al. 2021. Regulatory effects of HIF-1α and HO-1 in hypoxia-induced proliferation of pulmo-nary arterial smooth muscle cells in yak[J]. Cellular Sig-nalling, 87: 110140-110151.
[24] Zhao S, Gao J, Li J, Wang S, et al. 2021. PD-L1 regulates in-flammation in LPS-induced lung epithelial cells and vas-cular endothelial cells by interacting with the HIF-1α signaling pathway[J]. Inflammation, 44(5): 1969-1981.