Characteristic of a group of genes with low level of expression in the pancreas of rats under conditions of multi-day intermittent hypoxia influence

Authors

DOI:

https://doi.org/10.14739/2310-1237.2024.1.301114

Keywords:

pancreas, hypoxia, Bhlhe40 genes, Ctsa, Hif1a, Lox, Slc16a3, insulin, glucose, carbohydrate metabolism, fat metabolism, insulin resistance

Abstract

In modern medical science great attention is paid to the clarification of the molecular mechanisms, which are the basis of adaptation to environmental factors of unusual origin and/or extraordinary strength.

The aim of the study is to determine the features of a group of genes with low expression level, associated with hypoxia in the pancreas of Wistar rats under conditions of intermittent hypoxia.

Materials and methods. The study was conducted on 10 white, sexually mature Wistar rats, which were divided into 2 groups (5 animals in each). Animals of group 1 were part of the control (intact) group. The animals of the 2nd group were subjected to hypoxic training according to the following scheme: for 15 days, 6 hours daily, namely on days 1–5 they simulated an ascent to a height of one to five kilometers above sea level under the conditions of a barometer, and the last 10 days 6 km above the sea level.

To analyze gene expression, we used the polymerase chain reaction method with real-time reverse transcription (PCR) CFX-96 Touch™ (Bio-Rad, USA) and the RT2 Profiler™ PCR Array Rat Hypoxia Signaling Pathway kit (QIAGEN, Germany), where 84 genes were the subject of research in experimental animals.

Results. According to the results of the PCR study of genes in the pancreas samples of intact animals and animals exposed to hypoxic training, it was established that out of 84 genes associated with hypoxia, a group of 5 genes with a low expression level (∆∆Ct < 30) was found. This pattern includes Bhlhe40 genes, Ctsa, Hif1a, Lox, and Slc16a3, the expression of which is statistically reduced. Thus, compared to the level of their expression in intact animals, the expression of Bhlhe40 decreased by 2.59 times, Ctsa by 6.02 times, Hif1a by 3.85 times, Lox by 3.01 times, and Slc16a3 by 2.40 times.

Conclusions. Intermittent hypoxia reduces the expression of the Bhlhe40 gene by 2.59 times, which can be considered as an element of adaptation of cells to a low level of oxygen and modulation of genetic programs. The decrease in Ctsa gene expression by 6.02, Hif1a by 3.85, and Lox by 3.01 times during intermittent hypoxia demonstrates, that these effects can be used as sanogenic factors in insulin resistance and type 2 diabetes. The 2.40-fold decreased expression level of Slc16a3 is probably an element of metabolic adaptation and adaptation of the metabolic pathway of cells to hypoxia conditions.

Author Biographies

T. V. Ivanenko, Zaporizhzhia State Medical and Pharmaceutical University, Ukraine

MD, PhD, DSc, Assosiated Professor of the Department of Pathological Physiology with Course of Normal Physiology

Yu. M. Kolesnyk, Zaporizhzhia State Medical and Pharmaceutical University, Ukraine

MD, PhD, DSc, Professor of the Department of Pathological Physiology with the Course of Normal Physiology, Rector of Zaporizhzhia State Medical and Pharmaceutical University, Honored Science and Technology Figure of Ukraine

A. V. Abramov, Zaporizhzhia State Medical and Pharmaceutical University, Ukraine

MD, PhD, DSc, Professor of the Department of Pathological Physiology with the Course of Normal Physiology

References

Hoang M, Joseph JW. The role of α-ketoglutarate and the hypoxia sensing pathway in the regulation of pancreatic β-cell function. Islets. 2020;12(5):108-19. doi: https://doi.org/10.1080/19382014.2020.1802183

Ivanenko TV, Kolesnyk YM, Abramov AV. [Experimental study of the pattern of genes activated by multi-day intermittent hypoxia in the rat pancreas]. Pathologia. 2023;20(3):218-21. Ukrainian. doi: https://doi.org/10.14739/2310-1237.2023.3.292536

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402-8. doi: https://doi.org/10.1006/meth.2001.1262

Ivanenko TV, Kolesnyk YM, Abramova TV. Analiz endokrynnoho statusu ta rivnia ekspresii bilkiv apoptozu i proliferatsii v pankreatychnykh ostrivtsiakh shchuriv z eksperymentalnym tsukrovym diabetom pislia zakinchennia pereryvchastykh hipoksychnykh trenuvan. Patolohiia, reabilitatsiia, adaptatsiia. 2017;15(2):17-20. Ukrainian.

Ivanenko TV. Vliyanie gipoksicheskikh trenirovok na tsitoarkh-itektoniku pankreaticheskikh ostrovkov pri eksperimentalnom sakharnom diabete. Pathologia. 2015;(1 Suppl):52-3. Russian.

Ivanenko TV, Abramov AV, Kolesnyk YM, Zhulinskyi VO, Kovalov MM. Stymuliatsiia funktsionalnoho stanu beta-klityn pankreatychnykh ostrivtsiv bahatodennoiu pereryvchastoiu hipoksiieiu. Zdobutky klinichnoi i eksperymentalnoi medytsyny. 2013;(2):246. Ukrainian.

Bottalico LN, Weljie AM. Cross-species physiological interactions of endocrine disrupting chemicals with the circadian clock. Gen Comp Endocrinol. 2021;301:113650. doi: https://doi.org/10.1016/j.ygcen.2020.113650

Xu H, Xiang QY, Li S, Liu YS. High serum Bhlhe40 levels are associated with subclinical atherosclerosis in patients with type 2 diabetes mellitus: A cross-sectional study. Diab Vasc Dis Res. 2023;20(2):14791641231169246. doi: https://doi.org/10.1177/14791641231169246

Tsuyama T, Sato Y, Yoshizawa T, Matsuoka T, Yamagata K. Hypoxia causes pancreatic β-cell dysfunction and impairs insulin secretion by activating the transcriptional repressor BHLHE40. EMBO Rep. 2023;24(8):e56227. doi: https://doi.org/10.15252/embr.202256227

Wang C, Liu W, Liu Z, Chen L, Liu X, Kuang S. Hypoxia Inhibits Myogenic Differentiation through p53 Protein-dependent Induction of Bhlhe40 Protein. J Biol Chem. 2015;290(50):29707-16. doi: https://doi.org/10.1074/jbc.M115.688671

Caciotti A, Catarzi S, Tonin R, Lugli L, Perez CR, Michelakakis H, et al. Galactosialidosis: review and analysis of CTSA gene mutations. Orphanet J Rare Dis. 2013;8:114. doi: https://doi.org/10.1186/1750-1172-8-114

Cowling RT. Cathepsin A Inhibitors to Treat Heart Disease: Much Potential, Many Questions. JACC Basic Transl Sci. 2019;4(3):345-7. doi: https://doi.org/10.1016/j.jacbts.2019.05.004

Toss MS, Miligy IM, Haj-Ahmad R, Gorringe KL, AlKawaz A, Mittal K, et al. The prognostic significance of lysosomal protective protein (cathepsin A) in breast ductal carcinoma in situ. Histopathology. 2019;74(7):1025-35. doi: https://doi.org/10.1111/his.13835

Linz D, Hohl M, Dhein S, Ruf S, Reil JC, Kabiri M, et al. Cathepsin A mediates susceptibility to atrial tachyarrhythmia and impairment of atrial emptying function in Zucker diabetic fatty rats. Cardiovasc Res. 2016;110(3):371-80. doi: https://doi.org/10.1093/cvr/cvw071

Ahmad J, Zubair M, Malik A, Siddiqui MA, Wangnoo SK. Cathepsin-D, adiponectin, TNF-α, IL-6 and hsCRP plasma levels in subjects with diabetic foot and possible correlation with clinical variables: a multicentric study. Foot (Edinb). 2012;22(3):194-9. doi: https://doi.org/10.1016/j.foot.2012.03.015

Gladek I, Ferdin J, Horvat S, Calin GA, Kunej T. HIF1A gene polymorphisms and human diseases: Graphical review of 97 association studies. Genes Chromosomes Cancer. 2017;56(6):439-52. doi: https://doi.org/10.1002/gcc.22449

Hua X, Hu G, Hu Q, Chang Y, Hu Y, Gao L, et al. Single-Cell RNA Sequencing to Dissect the Immunological Network of Autoimmune Myocarditis. Circulation. 2020;142(4):384-400. doi: https://doi.org/10.1161/CIRCULATIONAHA.119.043545

Xu F, Huang M, Chen Q, Niu Y, Hu Y, Hu P, et al. LncRNA HIF1A-AS1 Promotes Gemcitabine Resistance of Pancreatic Cancer by Enhancing Glycolysis through Modulating the AKT/YB1/HIF1α Pathway. Cancer Res. 2021;81(22):5678-91. doi: https://doi.org/10.1158/0008-5472.CAN-21-0281

Liu H, Zhou Y, Li Y, Gong Z. Important roles of Hif1a in maternal or adult BPA exposure induced pancreatic injuries. Sci Rep. 2023;13(1):11502. doi: https://doi.org/10.1038/s41598-023-38614-8

Wang N, Zhang C, Xu Y, Tan HY, Chen H, Feng Y. Berberine improves insulin-induced diabetic retinopathy through exclusively suppressing Akt/mTOR-mediated HIF-1α/VEGF activation in retina endothelial cells. Int J Biol Sci. 2021;17(15):4316-4326. doi: https://doi.org/10.7150/ijbs.62868

Jaśkiewicz M, Moszyńska A, Króliczewski J, Cabaj A, Bartoszewska S, Charzyńska A, et al. The transition from HIF-1 to HIF-2 during prolonged hypoxia results from reactivation of PHDs and HIF1A mRNA instability. Cell Mol Biol Lett. 2022;27(1):109. doi: https://doi.org/10.1186/s11658-022-00408-7

Saatci O, Kaymak A, Raza U, Ersan PG, Akbulut O, Banister CE, et al. Targeting lysyl oxidase (LOX) overcomes chemotherapy resistance in triple negative breast cancer. Nat Commun. 2020;11(1):2416. doi: https://doi.org/10.1038/s41467-020-16199-4

Li H, Zhu X, Cao X, Lu Y, Zhou J, Zhang X. Single-cell analysis reveals lysyl oxidase (Lox)+ fibroblast subset involved in cardiac fibrosis of diabetic mice. J Adv Res. 2023;54:223-37. doi: https://doi.org/10.1016/j.jare.2023.01.018

Yan M, Mehta JL, Hu C. LOX-1 and obesity. Cardiovasc Drugs Ther. 2011;25(5):469-76. doi: https://doi.org/10.1007/s10557-011-6335-3

Yan M, Mehta JL, Zhang W, Hu C. LOX-1, oxidative stress and inflammation: a novel mechanism for diabetic cardiovascular complications. Cardiovasc Drugs Ther. 2011;25(5):451-9. doi: https://doi.org/10.1007/s10557-011-6342-4

Tao Q, Li X, Zhu T, Ge X, Gong S, Guo J, Ma R. Lactate Transporter SLC16A3 (MCT4) as an Onco-Immunological Biomarker Associating Tumor Microenvironment and Immune Responses in Lung Cancer. Int J Gen Med. 2022;15:4465-74. doi: https://doi.org/10.2147/IJGM.S353592

Choi SH, Kim MY, Yoon YS, Koh DI, Kim MK, Cho SY, et al. Hypoxia-induced RelA/p65 derepresses SLC16A3 (MCT4) by downregulating ZBTB7A. Biochim Biophys Acta Gene Regul Mech. 2019;1862(8):771-85. doi: https://doi.org/10.1016/j.bbagrm.2019.06.004

Published

2024-04-22

How to Cite

1.
Ivanenko TV, Kolesnyk YM, Abramov AV. Characteristic of a group of genes with low level of expression in the pancreas of rats under conditions of multi-day intermittent hypoxia influence. Pathologia [Internet]. 2024Apr.22 [cited 2024Nov.9];21(1):23-7. Available from: http://pat.zsmu.edu.ua/article/view/301114

Issue

Section

Original research