Features of the lactate dehydrogenase isoenzymes spectrum in animal tissues and organs

N. V.  Kuzmina; R. D.  Ostapiv; D. D.  Ostapiv; P. I.  Golovach

doi:10.15421/0224124

N. V. Kuzmina Institute of Animal Biology NAAS
R. D. Ostapiv State Scientific Control Institute of Veterinary Preparations and Feed Additive
D. D. Ostapiv Institute of Animal Biology NAAS
P. I. Golovach Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies of Lviv

Keywords: energy metabolism, enzyme, polyacrylamide gel, skeletal muscles, heart, liver, kidneys, spleen, ovaries, rats, pigs, cows

Abstract

Activity and content of lactate dehydrogenase (LDH) isoenzymes is one of the objective criteria for assessing oxidative metabolism in organs and tissues and the physiological state of the whole organism. Lactate dehydrogenase (LDH) is a key enzyme of energy metabolism. LDH is a heterotetrameric protein consisting of subunits of two types H (cardiac) and M (muscular). This enzyme exists in five isoforms: LDH1, LDH2, LDH3, LDH4 and LDH5. Each of the isoforms is characterized by different physicochemical and catalytic properties and is found in most tissue cells. Content of LDH isoenzymes is not constant and depends on the intensity of oxidative metabolism in tissue cells and on the influence of various factors, including new therapeutic drugs and biogenic compounds. Therefore, the aim of the research was to study and establish the limits of isoenzyme content and LDH activity in individual tissues and organs of rats, pigs and cows to characterize and detect changes in the intensity of metabolism in the body of the animals under different physiological state. Tissue samples of rat, pig and cow organs were taken for research. Isoenzymes of lactate dehydrogenase were detected by staining according to Garbus in our modification. The content of isoenzymes (%) was determined by the TotalLab TL120 software, the activity of LDH in blood plasma (µkat/L) and in tissue homogenates (µkat/mg protein) – by the rate of NADH oxidation. Brain, spleen, liver, kidney, testis, muscle and blood plasma of rats and pigs were characterized by 5 catalytically active LDH isoenzymes. In muscle and liver tissues, LDH5 prevailed, and in the brain, testis and kidney – LDH1. LDH3 was most abundant isoenzyme in spleen tissue. It was found that the same activity of LDH in the tissues of the liver and testis of rats (lim 4.5–4.6 µkat/mg of protein) was provided by different LDH isoenzymes. In addition to tissue characteristics, the content of LDH isoenzymes depended on the physiological state of the organ. Thus, in the cow ovaries the activity of LDH was the same (2.8–3.0 µkat/mg of protein) under the physiological state of "follicular growth" and pathology – "hypofunction". LDH activity was lower by 0.6 and 1.0 µkat/mg of protein under "early" and "late" corpus luteum (2.0 and 2.2 µkat/mg of protein). The highest content of LDH1 was in the physiological state of "late corpus luteum", and LDH3, LDH4 and LDH5 – in "early corpus luteum". Content of LDH2 was high in the state of ovarian dysfunction – "hypofunction". Content of LDH2 was lower and had no statistical difference (19.8% and 22.1%) in other physiological states. Isoenzyme spectrum of LDH was characterized by tissue specificity. Tissues with a predominance of anaerobic processes (muscle and liver) were characterized by a high content of LDH5, and with a predominance of aerobic processes (brain, testis, kidneys) – by a high content of LDH1. The spleen was characterized by a high content of LDH3. It was established, that the same activity of the enzyme in the organs could be ensured by different content of LDH proteins. It has been proven that the content of individual LDH isoenzymes depended on the physiological state of the ovary.

References

Adeva-Andany, M., López-Ojén, M., Funcasta-Calderón, R., Ameneiros-Rodríguez, E., Donapetry-García, C., Vila-Altesor, M., & Rodríguez-Seijas, J. (2014). Comprehensive review on lactate metabolism in human health. Mitochondrion, 17, 76–100.

Arnold, P. K., & Finley, L. W. S. (2023). Regulation and function of the mammalian tricarboxylic acid cycle. The Journal of Biological Chemistry, 299(2), 102838.

Augoff, K., & Grabowski, K. (2004). Przydatność oznaczania dehydrogenazy mleczanowej w rozpoznawaniu chorób nowotworowych [Significance of lactate dehydrogenase measurements in diagnosis of malignancies]. Polski Merkuriusz Lekarski, 17(102), 644–647.

Aydin, S., Ugur, K., Aydin, S., Sahin, İ., & Yardim, M. (2019). Biomarkers in acute myocardial infarction: Current perspectives. Vascular Health and Risk Management, 15, 1–10.

Brancaccio, P., Lippi, G., & Maffulli, N. (2010). Biochemical markers of muscular damage. Clinical Chemistry and Laboratory Medicine, 48(6), 757–767.

Clark, S. W., & Yochim, J. M. (1971). Effect of ovarian steroids on lactic dehydrogenase activity in endometrium and myometrium of the rat uterus. Endocrinology, 89(2), 358–365.

Corrigan, R. (2011). Fundamentals of veterinary clinical pathology. 2nd edition. The Canadian Veterinary Journal, 52(2), 161.

Duka, T., Anderson, S. M., Collins, Z., Raghanti, M. A., Ely, J. J., Hof, P. R., Wildman, D. E., Goodman, M., Grossman, L. I., & Sherwood, C. C. (2014). Synaptosomal lactate dehydrogenase isoenzyme composition is shifted toward aerobic forms in primate brain evolution. Brain, Behavior and Evolution, 83(3), 216–230.

Dupont, J., & Scaramuzzi, R. J. (2016). Insulin signalling and glucose transport in the ovary and ovarian function during the ovarian cycle. The Biochemical Journal, 473(11), 1483–1501.

Garbus, J. (1971). Serum malate dehydrogenase isoenzymes as indicators of severe cellular injury. Clinica Chimica Acta, 35, 502–504.

Hsieh, Y. S., Yeh, M. C., Lin, Y. Y., Weng, S. F., Hsu, C. H., Huang, C. L., Lin, Y. P., & Han, A. Y. (2022). Is the level of serum lactate dehydrogenase a potential biomarker for glucose monitoring with type 2 diabetes mellitus? Frontiers in Endocrinology, 13, 1099805.

Hui, S., Ghergurovich, J. M., Morscher, R. J., Jang, C., Teng, X., Lu, W., Esparza, L. A., Reya, T., Le Zhan, Yanxiang Guo, J., White, E., & Rabinowitz, J. D. (2017). Glucose feeds the TCA cycle via circulating lactate. Nature, 551(7678), 115–118.

Jeong, S., Kim, J., Kim, Y., Kim, I., & Han, J. (2019). Serum lactate dehydrogenase activity and its isoenzyme patterns in patients with pectus excavatum. Journal of Thoracic Disease, 11(10), 4349–4356.

Józwik, M., Józwik, M., Teng, C., & Battaglia, F. C. (2007). Concentrations of monosaccharides and their amino and alcohol derivatives in human preovulatory follicular fluid. Molecular Human Reproduction, 13(11), 791–796.

Khan, A. A., Allemailem, K. S., Alhumaydhi, F. A., Gowder, S. J., & Rahmani, A. H. (2020). The biochemical and clinical perspectives of lactate dehydrogenase: An enzyme of active metabolism. Endocrine, Metabolic and Immune Disorders-Drug Targets, 20(6), 855–868.

Kosenko, M. V., Chukhriy, B. M., & Chaykovska, O. I. (2005). Vidtvorennya molochnoho poholiv’ya [Playing milk livestock]. Ukrainskiye Tekhnologii, L’viv (in Ukrainian).

Ksendzovsky, A., Bachani, M., Altshuler, M., Walbridge, S., Mortazavi, A., Moyer, M., Chen, C., Fayed, I., Steiner, J., Edwards, N., Inati, S. K., Jahanipour, J., Maric, D., Heiss, J. D., Kapur, J., & Zaghloul, K. A. (2022). Chronic neuronal activation leads to elevated lactate dehydrogenase A through the AMP-activated protein kinase/hypoxia-inducible factor-1α hypoxia pathway. Brain Communications, 5(1), fcac298.

Kuzmina, N. V., Ostapiv, D. D., Kosenko, Y. M. (2020). Metodyky doslidzhen’ spektru izozymiv enerhetychnoho obminu i antyoksydantnoho zakhystu [Methods of research of the spectrum of isosisms of energy metabolism and antioxidant protection]. SSRCI of Veterinary Medical Products and Fodder Additives, Lviv (in Ukrainian).

Levchenko, V. I., & Vlizlo, V. V. (2019). Veterynarna klinichna biokhimiia [Veterinary clinical biochemistry]. BSAU, Bila Tserkva (in Ukrainian).

Nagy, O., Tóthová, C., & Chovanová, F. (2020). Clinical and diagnostic significance of lactate dehydrogenase and its isoenzymes in animals robert klein. Veterinary Medicine Internaion, 15, 5346483.

Niu, P., Jiang, J., Liu, K., Zhou, X., Wang, S., Xu, T., Wang, T., Li, Y., Yang, Q., & Liu, T. (2024). Hollow microsphere integrated optofluidic immunochip for myocardial infarction biomarker microanalysis. Biosensors and Bioelectronics, 248, 115970.

Ostapiv, R. D., Humenyuk, S. L., & Manko, V. V. (2015). Аctivity and isozyme content of lactate dehydrogenase under long-term oral taurine administration to rats. Ukranian Biochemical Journal, 87(4), 54–62.

Radaković, M., Borozan, S., Djelić, N., Ivanović, S., Miladinović, D. Ć., Ristanić, M., Spremo-Potparević, B., & Stanimirović, Z. (2018). Nitroso-oxidative stress, acute phase response, and cytogenetic damage in Wistar rats treated with adrenaline. Oxidative Medicine and Cellular Longevity, 2018, 1805354.

Robin, A. Y., Brochier-Armanet, C., Bertrand, Q., Barette, C., Girard, E., & Madern, D. (2023). Deciphering evolutionary trajectories of lactate dehydrogenases provides new insights into allostery. Molecular Biology and Evolution, 40(10), msad223.

Schurr, A. (2006). Lactate: The ultimate cerebral oxidative energy substrate? Journal of Cerebral Blood Flow and Metabolism, 26(1), 142–152.

Szrejder, M., Typiak, M., Pikul, P., Audzeyenka, I., Rachubik, P., Rogacka, D., Narajczyk, M., & Piwkowska, A. (2023). Role of L-lactate as an energy substrate in primary rat podocytes under physiological and glucose deprivation conditions. European Journal of Cell Biology, 102(2), 151298.

Takehana, K., Adachi, M., Ishikawa, S., & Yamagishi, N. (2023). Agarose gel electrophoresis pattern of serum creatine kinase and lactate dehydrogenase isoenzymes in zoo-managed Asian elephants (Elephas maximus). The Journal of Veterinary Medical Science, 85(5), 578–583.

Tokinoya, K., Ishikura, K., Yoshida, Y., Ra, S. G., Sugasawa, T., Aoyagi, A., Nabekura, Y., Takekoshi, K., & Ohmori, H. (2020). LDH isoenzyme 5 is an index of early onset muscle soreness during prolonged running. The Journal of Sports Medicine and Physical Fitness, 60(7), 1020–1026.

Vanithakumari, G., Govindarajulu, P., & Srinivasan, N. (1980). Hormonal influences on the oviducal lactate dehydrogenase and its isoenzymes. Indian Journal of Physiology and Pharmacology, 24(4), 335–340.

Vlizlo, V. V., Fedoruk, R. S., & Ratych, I. B. (Eds.). (2012). Laboratorni metody doslidzhen’ u biolohiyi, tvarynnytstvi ta veterynarniy medytsyni [Laboratory research methods in biology, animal husbandry and veterinary medicine]. Spolom, Lviv (in Ukrainian).

Ying, S., Wang, Z., Wang, C., Nie, H., He, D., Jia, R., Wu, Y., Wan, Y., Zhou, Z., Yan, Y., Zhang, Y., & Wang, F. (2011). Effect of different levels of short-term feed intake on folliculogenesis and follicular fluid and plasma concentrations of lactate dehydrogenase, glucose, and hormones in Hu sheep during the luteal phase. Reproduction, 142(5), 699–710.

Zhou, Y., Qi, M., & Yang, M. (2020). Current status and future perspectives of lactate dehydrogenase detection and medical implications: A review. Biosensors, 12(12), 1145.