高海拔环境抑制高原鼢鼠和高原鼠兔的胆汁酸代谢

doi:10.16829/j.slxb.150805

摘要/Abstract

摘要： 高原鼢鼠和高原鼠兔是青藏高原特有动物，低氧寒冷的环境影响其生理活动。胆汁酸是一种具有两亲特性的固醇类衍生物，不仅有利于脂溶性营养成分和维生素吸收，而且作为信号激素参与组织器官的糖脂代谢和能量代谢，具有调节功能，与动物适应环境有关。本文应用靶向代谢组学和荧光PCR定量方法，以Sprague-Dawley (SD)大鼠为对照，比较分析了SD大鼠、高原鼢鼠(Myospalax baileyi)和高原鼠兔(Ochotona curzoniae)血清中胆汁酸的组成和含量以及肝脏中初级胆汁酸合成关键酶基因的表达水平。结果表明，高原鼢鼠和高原鼠兔血清总胆汁酸含量均显著低于SD大鼠；随海拔升高，两种高原动物血清胆汁酸含量均显著下降，高原鼢鼠显著低于高原鼠兔，肝脏中初级胆汁酸合成关键酶基因CYP7A1、CYP8B1、CYP27A1和CYP7B1表达水平均显著下降；SD大鼠初级胆汁酸主要通过经典途径合成，而高原鼢鼠和高原鼠兔初级胆汁酸主要通过替代途径合成，提示高海拔环境抑制胆汁酸合成，低氧可能是主要的抑制因子。两种高原动物血清胆汁酸以非12-羟基胆汁酸和非结合型次级胆汁酸为主，结合型胆汁酸以甘氨酸结合型为主。特别是在高原鼢鼠血清中，石胆酸(lithocholic acid,LCA)类胆汁酸含量显著高于高原鼠兔和SD大鼠，这可能与其适应高耗能的地下挖掘生活有关。总之，高原鼢鼠和高原鼠兔初级胆汁酸合成受低氧抑制且以替代途径为主，这可能是它们长期适应高原环境的结果。

关键词: 高原鼢鼠, 高原鼠兔, 血清, 胆汁酸, 低氧, 胆汁酸合成关键酶基因

Abstract: The plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae), which are native to the Qinghai-Xizang Plateau, inhabit hypoxic meadows that pose unique physiological challenges. Bile acids are amphipathic steroid metabolites that not only facilitate intestinal absorption of lipids and fat-soluble vitamins, but also act as hormones capable of reaching virtually every organ to regulate glycometabolism, lipid metabolism, and energy homeostasis, which are related to the adaptation of animals to the environments. In this study, the composition and contents of bile acids in the serum of plateau zokors and plateau pikas were analyzed by targeting metabolomics. The Sprague-Dawley (SD) rats were used as control and the expression levels of the key genes of bile acids anabolism were determined by real-time PCR. The results showed that the total bile acid contents in the serum of plateau zokors and plateau pikas decreased markedly than that of SD rats. With increasing altitude, the bile acid contents of zokors and pikas decreased significantly, and that of zokors reduced remarkably compared to that of pikas. The expression levels of the key genes of primary bile acids including CYP7A1, CYP8B1, CYP27A1, and CYP7B1 decreased significantly in livers of zokors and pikas. The bile acids of SD rats were synthesized by the classic or neutral bile acid pathway, but that of zokors and pikas were produced by the alternative or acidic pathway. These results suggest that high-altitude environments inhibit the anabolism of primary bile acids and that hypoxia might be the main inhibiting factor. The no-12-OH bile acids and the unconjugated secondary bile acids were the main products and the glycine conjugated bile acids were the main bile acids in the serum of zokors and pikas. Particularly, the contents of lithocholic acid (LCA) bile acids were significantly higher than that of pikas and SD rats in serum which might be related to its adaptation to the higher energy expenditure of burrowing activities. In conclusion, the alternative pathway was the dominant pathway of primary bile acids in the liver of zokors and pikas, which might be a result of long-time adaptation to the highland environments.

Key words: Plateau zokor (Myospalax baileyi), Plateau pika (Ochotona curzoniae), Serum, Bile acids, Hypoxia, Key enzyme genes for bile acid synthesis

中图分类号:

Q955

张佳钰, 安志芳, 王志洁, 陈晓琦, 魏登邦. 高海拔环境抑制高原鼢鼠和高原鼠兔的胆汁酸代谢[J]. 兽类学报, 2023, 43(5): 553-567.

ZHANG Jiayu, AN Zhifang, WANG Zhijie, CHEN Xiaoqi, WEI Dengbang. Inhibition of bile acid metabolism in plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae) in high-altitude environments[J]. ACTA THERIOLOGICA SINICA, 2023, 43(5): 553-567.

参考文献

Adak A, Maity C, Ghosh K, Mondal K C. 2014. Alteration of predominant gastrointestinal flora and oxidative damage of large intestine under simulated hypobaric hypoxia. Zeitschrift fur Gastroenterologie, 52 (2):180-186.
An Z F, Wei L N, Xu B, Wang Z J, Gao C H, Li J M, Wei L, Qi D L, Shi P, Zhang T Z, Wei D B. 2022. A homotetrameric hemoglobin expressed in alveolar epithelial cells increases blood oxygenation in high-altitude plateau pika (Ochotona curzoniae). Cell Reports, 41 (1):111446.
Bensalem A, Murtaza B, Hichami A, Khan A S, Oulamara H, Merlen G, Berrichi M, Agli A, Tordjmann T, Khan N A. 2020. Bile acid receptor TGR5 is critically involved in preference for dietary lipids and obesity. The Journal of Nutritional Biochemistry, 76:108298.
Cai Z Y, Wang L Y, Song X Y, Tagore S, Li X F, Wang H H, Chen J R, Li K X, Frenkel Z, Gao D H, Frenkel-Morgenstern M, Zhang T Z, Nevo E. 2018. Adaptive transcriptome profiling of subterranean zokor, Myospalax baileyi, to high-altitude stresses in Tibet. Scientific Reports, 8 (1):1-11.
Carino A, Cipriani S, Marchianò S, Biagioli M, Scarpelli P, Zampella A, Monti M C, Fiorucci S. 2017. Gpbar1 agonism promotes a Pgc-1α-dependent browning of white adipose tissue and energy expenditure and reverses diet-induced steatohepatitis in mice. Scientific Reports, 7 (1):1-13.
Castellanos-Jankiewicz A, Guzmán-Quevedo O, Fenelon V S, Allard C, Quemener S, Guinot V, Hennuyer N, Perino A, Duveau A, Maitre M, Leste-Lasserre T, Clark S, Dupuy N, Cannich A, Gonzales D, Deprez B, Mithieux G, Dombrowicz D, Bäckhed F, Prevot V, Marsicano G, Staels B, Schoonjans K, Cota D. 2021. Hypothalamic bile acid-TGR5 signaling protects from obesity. Cell Metabolism, 33 (7):1483-1492.
Chaudhari S N, Luo J N, Harris D A, Aliakbarian H, Yao L, Paik D, Subramaniam R, Adhikari A A, Vernon A H, Kiliç A, Weiss S T, Huh J R, Sheu E G, Devlin A S. 2021. A microbial metabolite remodels the gut-liver axis following bariatric surgery. Cell Host & Microbe, 29 (3):408-424.
Chen X Q, An Z F, Wei L N, Zhang J Y, Li J M, Wang Z J, Gao C H, Wei D B. 2022. Vitamin D3 metabolic enzymes in plateau zokor(Myospalax baileyi) and plateau pika (Ochotona curzoniae):expression and response to Hypoxia. Animals, 12 (18):2371.
Fan C, Zhang L Z, Jia S G, Tang X J, Fu H B, Li W J, Liu C F, Zhang H, Cheng Q, Zhang Y M. 2022. Seasonal variations in the composition and functional profiles of gut microbiota reflect dietary changes in plateau pikas. Integrative Zoology, 17 (3):379-395.
Fan N C, Shi Y Z. 1982. A revision of the zokors of subgenus Eospalax. Acta Theriologica Sinica, 2 (2):183-199. (in Chinese)
Feng Z J, Zheng C L. 1985. Studies on the pikas (genus:Ochotona) of China-Taxonomic notes and distribution. Acta Theriologica Sinica, 5 (4):269-290. (in Chinese)
Funabashi M, Grove T L, Wang M, Varma Y, McFadden M E, Brown L C, Guo C J, Higginbottom S, Almo S C, Fischbach M A. 2020. A metabolic pathway for bile acid dehydroxylation by the gut microbiome. Nature, 582 (7813):566-570.
Gadaleta R M, Moschetta A. 2019. Metabolic messengers:fibroblast growth factor 15/19. Nature Metabolism, 1 (6):588-594.
Goodwin B, Jones S A, Price R R, Watson M A, McKee D D, Moore L B, Galardi C, Wilson J G, Lewis M C, Roth M E, Maloney P R, Willson T M, Kliewer S A. 2000. A regulatory cascade of the nuclear receptors FXR, SHP-1, and LRH-1 represses bile acid biosynthesis. Molecular Cell, 6 (3):517-526.
Gribble F M, Reimann F. 2019. Function and mechanisms of enteroendocrine cells and gut hormones in metabolism. Nature Reviews. Endocrinology, 15 (4):226-237.
Jia W, Wei M, Rajani C, Zheng X. 2021. Targeting the alternative bile acid synthetic pathway for metabolic diseases. Protein & Cell, 12(5):411-425.
Li H, Zhou R, Zhu J X, Huang X D, Qu J P. 2019. Environmental filtering increases with elevation for the assembly of gut microbiota in wild pikas. Microbial Biotechnology, 12 (5):976-992.
Liu R H. 1995. Classification and geographic zoning of Chinese zokor. Territory & Natural Resources Study, 3:54-55. (in Chinese)
Lund M L, Sorrentino G, Egerod K L, Kroone C, Mortensen B, Knop F K, Reimann F, Gribble F M, Drucker D J, de Koning E J P, Schoonjans K, Bäckhed F, Schwartz T W, Petersen N. 2020. Lcell differentiation is induced by bile acids through GPBAR1 and paracrine GLP-1 and serotonin signaling. Diabetes, 69 (4):614-623.
Makishima M, Lu T T, Xie W, Whitfield G K, Domoto H, Evans R M, Haussler M R, Mangelsdorf D J. 2002. Vitamin D receptor as an intestinal bile acid sensor. Science, 296 (5571):1313-1316.
Nehring J A, Zierold C, De Luca H F. 2007. Lithocholic acid can carry out in vivo functions of vitamin D. Proceedings of the National Academy of Sciences of the United States of America, 104(24):10006-10009.
Pathak P, Xie C, Nichols R G, Ferrell J M, Boehme S, Krausz K W, Patterson A D, Gonzalez F J, Chiang J Y. 2018. Intestine farnesoid X receptor agonist and the gut microbiota activate G-protein bile acid receptor-1 signaling to improve metabolism. Hepatology, 68 (4):1574-1588.
Perino A, Schoonjans K. 2022. Metabolic messengers:bile acids. Nature Metabolism, 4 (4):416-423.
Sasaki T, Watanabe Y, Kuboyama A, Oikawa A, Shimizu M, Yamauchi Y, Sato R. 2021. Muscle - specific TGR5 overexpression improves glucose clearance in glucose-intolerant mice. The Journal of Biological Chemistry, 296:100131.
Shao Y, Li J X, Ge R L, Zhong L, Irwin D M, Murphy R W, Zhang Y P. 2015. Genetic adaptations of the plateau zokor in high-elevation burrows. Scientific Reports, 5 (1):17262.
Shi Y Z, Fan N C. 1980. Rodent Damage and Its Prevention on Grassland. Xining:Qinghai People's Publishing House, 67-105. (in Chinese)
Stefano F, Eleonora D, Adriana C, Angela Z, Michele B. 2021. Bile acids and their receptors in metabolic disorders. Progress in Lipid Research, 82:101094.
Velazquez-Villegas L A, Perino A, Lemos V, Zietak M, Nomura M, Pols T W H, Schoonjans K. 2018. TGR5 signalling promotes mitochondrial fission and beige remodelling of white adipose tissue. Nature Communications, 9 (1):245.
Wahlström A, Sayin S I, Marschall H U, Bäckhed F. 2016. Intestinal crosstalk between bile acids and microbiota and its impact on host metabolism. Cell Metabolism, 24 (1):41-50.
Wang D M, Doestzada M, Chen L, Andreu-Sánchez S, van den Munckhof I C, Augustijn H E, Koehorst M, Ruiz-Moreno A J, Bloks V W, Riksen N P, Rutten J H W, Joosten L A B, Netea M G, Wijmenga C, Zhernakova, Kuipers F, Fu J Y. 2021. Characterization of gut microbial structural variations as determinants of human bile acid metabolism. Cell Host & Microbe, 29 (12):1802-1814.
Zeng J X, Wang Z W, Shi Z X. 1984. Metabolic characteristics and some physiological parameters of mole rat Myospalax baileyi in alpine area. Acta Biologica Plateau Sinica, 3:163-171. (in Chinese)
Zhang T, Chen J, Zhang J, Guo Y T, Zhou X, Li M W, Zheng Z Z, Zhang T Z, Murphy R W, Nevo E, Shi P. 2021. Phenotypic and genomic adaptations to the extremely high elevation in plateau zokor (Myospalax baileyi). Molecular Ecology, 30 (22):5765-5779.
Zhang X L, Xu T W, Wang X G, Geng Y Y, Liu H J, Hu L Y, Zhao N, Kang S P, Zhang W M, Xu S X. 2020. The effect of transitioning between feeding methods on the gut microbiota dynamics of yaks on the Qinghai-Tibet Plateau. Animals, 10 (9):1641.
冯祚建, 郑昌琳. 1985. 中国鼠兔属 (Ochotona) 的研究:分类与分布. 兽类学报, 5 (4):269-290.
刘仁华. 1995. 中国鼢鼠的分类及地理区划. 国土与自然资源研究, 3:54-56.
施银柱, 樊乃昌. 1980. 草原鼠害及其防治. 西宁:青海人民出版社, 67-105.
曾缙祥, 王祖望, 师治贤. 1984. 高山地区高原鼢鼠的代谢特点及若干生理指标的观察. 高原生物学集刊, 3:163-171.
樊乃昌, 施银柱. 1982. 中国鼢鼠 (EOSPALAX) 亚属分类研究. 兽类学报, 2 (2):183-199.

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