anatomyЖурнал анатомии и гистопатологииJournal of Anatomy and Histopathology2225-7357N.N. Burdenko Voronezh State Medical University10.18499/2225-7357-2023-12-4-38-46anatomy-1836Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL PAPERSОсобенности экспрессии белка Клото в почках крыс при экспериментальной гипергликемии на фоне фармакологической коррекции производными ГАМКFeatures of Klotho Protein Expression in Rat Kidneys in Experimental Hyperglycemia Against the Background of Pharmacological Correction with GABA Derivativeshttps://orcid.org/0000-0003-4525-3688КавалероваД. А.KavalerovaD. A.

Кавалерова Дарья Алевтиновна – старший преподаватель кафедры биологии

 пл. Павших борцов, 1, Волгоград, 400066

Dar’ya A. Kavalerova – senior teacher of the Department of Biology

pl. Pavshikh bortsov, 1, Volgograd, 400066

dareznikova@yandex.ruhttps://orcid.org/0000-0002-8612-6186СнигурГ. Л.SnigurG. L.

Снигур Григорий Леонидович – д-р мед. наук, доцент, заведующий кафедрой биологии

 пл. Павших борцов, 1, Волгоград, 400066

Grigorii L. Snigur – Doct. Sci. (Med.), Associate Professor, Head of the Department of Biology

pl. Pavshikh bortsov, 1, Volgograd, 400066

sgrigoryl@mail.ruhttps://orcid.org/0000-0002-7770-5995СуринС. С.SurinS. S.

Сурин Святослав Сергеевич – старший преподаватель кафедры биологии

 пл. Павших борцов, 1, Волгоград, 400066

Svyatoslav S. Surin - senior teacher of the Department of Biology

pl. Pavshikh bortsov, 1, Volgograd, 400066

svyatoslavsurin@gmail.comhttps://orcid.org/0000-0001-7574-3923ТюренковИ. Н.TyurenkovI. N.

Тюренков Иван Николаевич – д-р мед. наук, профессор, член-корр. РАН, заведующий кафедрой фармакологии и фармации

 пл. Павших борцов, 1, Волгоград, 400066

Ivan N. Tyurenkov – Doct. Sci. (Med.), Professor, Corresponding member of RAS, Head of the Department of Pharmacology and Pharmacy of Institute of Continuing Medical and Pharmacological Education

pl. Pavshikh bortsov, 1, Volgograd, 400066

fibfuv@mail.ruhttps://orcid.org/0000-0003-4694-3066БакулинД. А.BakulinD. A.

Бакулин Дмитрий Александрович – канд. мед. наук, старший научн. сотр. лаборатории фармакологии сердечно-сосудистых средств

 пл. Павших борцов, 1, Волгоград, 400066

Dmitrii A. Bakulin – Cand. Sci. (Med.), senior researcher of Laboratory of Pharmacology of Cardiovascular Drugs

pl. Pavshikh bortsov, 1, Volgograd, 400066

mbfdoc@gmail.comhttps://orcid.org/0000-0003-0143-9245СахароваЭ. Ю.SakharovaE. Yu.

Сахарова Элина Юрьевна – канд. мед. наук, доцент кафедры биологии

 пл. Павших борцов, 1, Волгоград, 400066

Elina Yu. Sakharova – Cand. Sci. (Med.), Associate Professor of the Department of Biology

pl. Pavshikh bortsov, 1, Volgograd, 400066

saharova_elina@mail.ruВолгоградский государственный медицинский университетVolgograd State Medical University2023180120241243846Copyright © Кавалерова Д.А., Снигур Г.Л., Сурин С.С., Тюренков И.Н., Бакулин Д.А., Сахарова Э.Ю., 20242024Кавалерова Д.А., Снигур Г.Л., Сурин С.С., Тюренков И.Н., Бакулин Д.А., Сахарова Э.Ю.Kavalerova D.A., Snigur G.L., Surin S.S., Tyurenkov I.N., Bakulin D.A., Sakharova E.Y.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://anatomy.elpub.ru/jour/article/view/1836

Цель – выявить особенности экспрессии белка Клото в эпителии извитых канальцев почки при длительной экспериментальной гипергликемии и фармакологической коррекции производными ГАМК. Материал и методы. Исследование проведено на 50 беспородных крысах-самцах (исходная масса: 330,0–360,0 г) со стрептозотоциновым диабетом длительностью 6 мес. Через 6 мес. после инъекции стрептозотоцина (60 мг/кг) в исследование включали животных с уровнем гликемии ≥ 15 ммоль/л (после 4-часовой пищевой депривации), затем в течение 30 дней перорально вводили производные ГАМК (аминалон, мефаргин, сукцикард). Далее оценивали протеинурию и концентрацию креатинина в сыворотке крови. После эвтаназии ткань почек фиксировали в забуференном нейтральном формалине и исследовали с помощью методов иммунофлуоресцентной микроскопии. Интенсивность свечения оценивали на основании визуально-аналоговой шкалы. Результаты. При определении уровня суточной протеинурии и уровня креатинина в сыворотке крови в группе с хронической гипергликемией без лечения отмечалось значительное повышение содержания белка в суточной моче и уровня креатинина в сыворотке крови (p<0,05). При проведении иммунофлуоресцентного анализа в группе животных с хронической гипергликемией без лечения отмечалось значительное снижение экспрессии белка Клото по сравнению с группой интактных животных. В группе, курсом получавшей производное ГАМК сукцикард, отмечалось улучшение функции почек, сопровождающееся достоверным увеличением (p≤0,0001) площади Клотопозитивного материала в эпителии извитых канальцев почки по отношению к группе животных с длительной гипергликемией без лечения. Заключение. Длительная гипергликемия вызывает выраженное нарушение функции почек у крыс со стрептозотоциновым диабетом: повышение уровня протеинурии и креатинина в сыворотке крови сопровождается снижением экспрессии белка Клото в почках. Производное ГАМК сукцикард при курсовом введении улучшает функцию почек, и данный эффект сопровождается повышением экспрессии белка Клото.

The aim is to identify the features of Klotho protein expression in the epithelium of the convoluted tubules of the kidney during prolonged experimental hyperglycemia and pharmacological correction with GABA derivatives. Material and methods. The study used 50 mongrel male rats (initial weight: 330,0–360,0 g) with streptozotocin diabetes lasting 6 months. 6 months after the injection of streptozotocin (60 mg/kg), animals with a glycemic level of ≥ 15 mmol/l were included in the study (after 4 hours of food deprivation), then GABA derivatives (aminalon, mefargine, succicard) were administered orally for 30 days. Proteinuria and serum creatinine concentrations were evaluated. After euthanasia, kidney tissue was fixed in buffered neutral formalin and examined using immunofluorescence microscopy. The intensity of the glow was evaluated based on a visual-analog scale. Results. In the group with chronic hyperglycemia without treatment, there was a significant increase in the protein content in daily urine and serum creatinine levels (p<0,05). In the group of animals with chronic hyperglycemia without treatment, there was a significant decrease in the expression of Klotho protein compared to the group of intact animals during immunofluorescence analysis. In the group treated with the GABA derivative succicard, there was an improvement in the functional state of the kidneys, accompanied by a significant increase (p≤0,0001) in the area of Klotho-positive material in the epithelium of the convoluted tubules of the kidney in relation to the group of animals with prolonged hyperglycemia without treatment. Conclusion. Prolonged hyperglycemia causes severe renal dysfunction in rats with streptozotocin diabetes: an increase the proteinuria and serum creatinine levels is accompanied by a decrease in the expression of Klotho protein in the kidneys. The GABA derivative succicard with course administration improves kidney function and this effect is accompanied by an increase in the expression of Klotho protein.

белок Клотогипергликемияпочкаизвитые канальцыпроизводные ГАМКKlotho proteinhyperglycemiakidneyconvoluted tubulesGABA derivativesработа выполнена при финансовой поддержке гранта РНФ №21-15-00192 от 19.04.2021ReferencesНестерова А.А., Глинка Е.Ю., Тюренков И.Н., Перфилова В.Н. Белок Клото – универсальный регулятор физиологических процессов в организме. Успехи физиологических наук. 2020;51(2):88–104. doi: 10.31857/S0301179820020083Nesterova A.A., Glinka E.Yu., Tyurenkov I.N., Perfilova V.N. Protein Klotho – Universal Regulator of Physiological Processes in the Organism. Progress in Physiological Science. 2020;51(2):88–104 (In Russ.). doi: 10.31857/S0301179820020083Спасов А.А., Воронкова М.П., Снигур Г.Л. Методические рекомендации по доклиническому изучению пероральных лекарственных средств для лечения сахарного диабета. Руководство по проведению доклинических исследований лекарственных средств: под ред. А.Н. Миронова. Ч.1. М.: Гриф и К, 2012; 670–84.Spasov A.A., Voronkova M.P., Snigur G.L. Metodicheskie rekomendatsii po doklinicheskomu izucheniyu peroral'nykh lekarstvennykh sredstv dlya lecheniya sakharnogo diabeta. Rukovodstvo po provedeniyu doklinicheskikh issledovanii lekarstvennykh sredstv: pod red. A.N. Mironova. Ch.1. M.: Grif i K, 2012; 670–84 (In Russ.).Тюренков И.Н., Файбисович Т.И., Дубровина М.А., Бакулин Д.А., Куркин Д.В. ГАМКергическая система в регуляции функционирования бета-клеток поджелудочной железы в условиях нормы и при сахарном диабете. Успехи физиологических наук. 2023;54(2):86–104. doi: 10.31857/s030117982302008xTyurenkov IN, Faibisovich TI, Dubrovina MA, Bakulin DA, Kurkin DV. Gabaergic System in The Regulation of the Functioning of Pancreas BetaCells in Normal Physiological Conditions And In Diabetes. Progress in Physiological Science. 2023;54(2):86–104. doi: 10.31857/s030117982302008x (In Russ.).Anders HJ, Huber TB, Isermann B, Schiffer M. CKD in diabetes: diabetic kidney disease versus nondiabetic kidney disease. Nature Reviews Nephrology. 2018 Apr 13;14(6):361–77. doi: 10.1038/s41581-018-0001-yAnders HJ, Huber TB, Isermann B, Schiffer M. CKD in diabetes: diabetic kidney disease versus nondiabetic kidney disease. Nature Reviews Nephrology. 2018 Apr 13;14(6):361–77. doi: 10.1038/s41581-018-0001-yBuendïa P, Ramı́rezR, Aljama P, Carracedo J. Klotho Prevents Translocation of NFκB. Vitamins and hormones. 2016 Jan 1;(101):119–50. doi: 10.1016/bs.vh.2016.02.005Buendïa P, Ramı́rezR, Aljama P, Carracedo J. Klotho Prevents Translocation of NFκB. Vitamins and hormones. 2016 Jan 1;(101):119–50. doi: 10.1016/bs.vh.2016.02.005Buendia-Roldan I, Machuca N, Mejía M, Maldonado M, Pardo A, Selman M. Lower levels of α-Klotho in serum are associated with decreased lung function in individuals with interstitial lung abnormalities. Scientific Reports. 2019 Jul 25;9(1):10801. doi: 10.1038/s41598-019-47199-0Buendia-Roldan I, Machuca N, Mejía M, Maldonado M, Pardo A, Selman M. Lower levels of α-Klotho in serum are associated with decreased lung function in individuals with interstitial lung abnormalities. Scientific Reports. 2019 Jul 25;9(1):10801. doi: 10.1038/s41598-019-47199-0Chen Z, Zhou Q, Liu C, Zeng Y, Yuan S. Klotho deficiency aggravates diabetes-induced podocyte injury due to DNA damage caused by mitochondrial dysfunction. International Journal of Medical Sciences. 2020;17(17):2763–72. doi: 10.7150/ijms.49690Chen Z, Zhou Q, Liu C, Zeng Y, Yuan S. Klotho deficiency aggravates diabetes-induced podocyte injury due to DNA damage caused by mitochondrial dysfunction. International Journal of Medical Sciences. 2020;17(17):2763–72. doi: 10.7150/ijms.49690Doi S, Zou Y, Osamu Togao, Pastor J, John G, Wang L, et al. Klotho Inhibits Transforming Growth Factor-β1 (TGF-β1) Signaling and Suppresses Renal Fibrosis and Cancer Metastasis in Mice. Journal of Biological Chemistry. 2011 Mar 11;286(10):8655–65. doi: 10.1074/jbc.M110.174037Doi S, Zou Y, Osamu Togao, Pastor J, John G, Wang L, et al. Klotho Inhibits Transforming Growth Factor-β1 (TGF-β1) Signaling and Suppresses Renal Fibrosis and Cancer Metastasis in Mice. Journal of Biological Chemistry. 2011 Mar 11;286(10):8655–65. doi: 10.1074/jbc.M110.174037Gheith O, Farouk N, Nampoory N, Halim MA, AlOtaibi T. Diabetic kidney disease: world wide difference of prevalence and risk factors. J Nephropharmacol. 2015 Oct 9;5(1):49-56. PMID: 28197499; PMCID: PMC5297507.Gheith O, Farouk N, Nampoory N, Halim MA, AlOtaibi T. Diabetic kidney disease: world wide difference of prevalence and risk factors. J Nephropharmacol. 2015 Oct 9;5(1):49-56. PMID: 28197499; PMCID: PMC5297507.Guo L, Tian J, Du H. Mitochondrial Dysfunction and Synaptic Transmission Failure in Alzheimer’s Disease. Journal of Alzheimer’s Disease. 2017 Apr 19;57(4):1071–86. doi: 10.3233/JAD-160702Guo L, Tian J, Du H. Mitochondrial Dysfunction and Synaptic Transmission Failure in Alzheimer’s Disease. Journal of Alzheimer’s Disease. 2017 Apr 19;57(4):1071–86. doi: 10.3233/JAD-160702Guo YL, Zhuang X, Huang Z, Zou J, Yang D, Hu X, et al. Klotho protects the heart from hyperglycemia-induced injury by inactivating ROS and NF-κB-mediated inflammation both in vitro and in vivo. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 2018 Jan 1;1864(1):238–51. doi: 10.1016/j.bbadis.2017.09.029Guo YL, Zhuang X, Huang Z, Zou J, Yang D, Hu X, et al. Klotho protects the heart from hyperglycemia-induced injury by inactivating ROS and NF-κB-mediated inflammation both in vitro and in vivo. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 2018 Jan 1;1864(1):238–51. doi: 10.1016/j.bbadis.2017.09.029Haruna Y, Kashihara N, Satoh M, Tomita N, Namikoshi T, Sasaki T, et al. Amelioration of progressive renal injury by genetic manipulation of Klotho gene. Proceedings of the National Academy of Sciences of the United States of America. 2007 Feb 13;104(7):2331–6. doi: 10.1073/pnas.0611079104Haruna Y, Kashihara N, Satoh M, Tomita N, Namikoshi T, Sasaki T, et al. Amelioration of progressive renal injury by genetic manipulation of Klotho gene. Proceedings of the National Academy of Sciences of the United States of America. 2007 Feb 13;104(7):2331–6. doi: 10.1073/pnas.0611079104Hua S, Liu Q, Li J, Fan M, Yan K, Ye D. Betaklotho in type 2 diabetes mellitus: From pathophysiology to therapeutic strategies. Reviews in Endocrine and Metabolic Disorders. 2021 Jun 13;22(4):1091–109. doi: 10.1007/s11154-021-09661-1Hua S, Liu Q, Li J, Fan M, Yan K, Ye D. Betaklotho in type 2 diabetes mellitus: From pathophysiology to therapeutic strategies. Reviews in Endocrine and Metabolic Disorders. 2021 Jun 13;22(4):1091–109. doi: 10.1007/s11154-021-09661-1Jiang W, Xiao T, Han W, Xiong J, He T, Liu Y, et al. Klotho inhibits PKCα/p66SHC-mediated podocyte injury in diabetic nephropathy. Molecular and Cellular Endocrinology. 2019 Aug 1;494:110490. doi: 10.1016/j.mce.2019.110490Jiang W, Xiao T, Han W, Xiong J, He T, Liu Y, et al. Klotho inhibits PKCα/p66SHC-mediated podocyte injury in diabetic nephropathy. Molecular and Cellular Endocrinology. 2019 Aug 1;494:110490. doi: 10.1016/j.mce.2019.110490Khanijou V, Zafari N, Coughlan MT, MacIsaac RJ, Ekinci EI. Review of potential biomarkers of inflammation and kidney injury in diabetic kidney disease. Diabetes/Metabolism Research and Reviews. 2022 Jul 11;38(6):3556. doi: 10.1002/dmrr.3556Khanijou V, Zafari N, Coughlan MT, MacIsaac RJ, Ekinci EI. Review of potential biomarkers of inflammation and kidney injury in diabetic kidney disease. Diabetes/Metabolism Research and Reviews. 2022 Jul 11;38(6):3556. doi: 10.1002/dmrr.3556Koch EAT, Nakhoul R, Nakhoul F, Nakhoul N. Autophagy in diabetic nephropathy: a review. International Urology and Nephrology. 2020 Jul 13;52(9):1705–12. doi: 10.1007/s11255-020- 02545-4Koch EAT, Nakhoul R, Nakhoul F, Nakhoul N. Autophagy in diabetic nephropathy: a review. International Urology and Nephrology. 2020 Jul 13;52(9):1705–12. doi: 10.1007/s11255-020- 02545-4Kuro-o M, Matsumura Y, Aizawa H, Kawaguchi H, Suga T, Utsugi T, et al. Mutation of the mouse klotho gene leads to a syndrome resembling ageing. Nature. 1997;390(6655):45–51. doi: 10.1038/36285 18. Kuro-o M. The Klotho proteins in health and disease. Nature Reviews Nephrology. 2018 Nov 19;15(1):27–44. doi: 10.1038/s41581-018-0078-3Kuro-o M, Matsumura Y, Aizawa H, Kawaguchi H, Suga T, Utsugi T, et al. Mutation of the mouse klotho gene leads to a syndrome resembling ageing. Nature. 1997;390(6655):45–51. doi: 10.1038/36285 18. Kuro-o M. The Klotho proteins in health and disease. Nature Reviews Nephrology. 2018 Nov 19;15(1):27–44. doi: 10.1038/s41581-018-0078-3Memmos E, Sarafidis P, Pateinakis P, Tsiantoulas A, Faitatzidou D, Giamalis P, et al. Soluble Klotho is associated with mortality and cardiovascular events in hemodialysis. BMC Nephrology. 2019 Jun 11;20(1):217. doi: 10.1186/s12882-019-1391-1Memmos E, Sarafidis P, Pateinakis P, Tsiantoulas A, Faitatzidou D, Giamalis P, et al. Soluble Klotho is associated with mortality and cardiovascular events in hemodialysis. BMC Nephrology. 2019 Jun 11;20(1):217. doi: 10.1186/s12882-019-1391-1Niture SK, Khatri R, Jaiswal AK. Regulation of Nrf2—an update. Free Radical Biology and Medicine. 2014 Jan;66:36–44. doi: 10.1016/j.freeradbiomed.2013.02.008Niture SK, Khatri R, Jaiswal AK. Regulation of Nrf2—an update. Free Radical Biology and Medicine. 2014 Jan;66:36–44. doi: 10.1016/j.freeradbiomed.2013.02.008Oh HJ, Nam BY, Wu M, Kim SH, Park J, Sukyung K, et al. Klotho plays a protective role against glomerular hypertrophy in a cell cycle-dependent manner in diabetic nephropathy. American Journal of Physiology-renal Physiology. 2018 Oct 1;315(4):F791–805. doi: 10.1152/ajprenal.00462.2017Oh HJ, Nam BY, Wu M, Kim SH, Park J, Sukyung K, et al. Klotho plays a protective role against glomerular hypertrophy in a cell cycle-dependent manner in diabetic nephropathy. American Journal of Physiology-renal Physiology. 2018 Oct 1;315(4):F791–805. doi: 10.1152/ajprenal.00462.2017Olejnik A, Franczak A, Krzywonos-Zawadzka A, Kałużna-Oleksy M, Bil-Lula I. The Biological Role of Klotho Protein in the Development of Cardiovascular Diseases. BioMed Research International. 2018;2018:5171945. doi: 10.1155/2018/5171945Olejnik A, Franczak A, Krzywonos-Zawadzka A, Kałużna-Oleksy M, Bil-Lula I. The Biological Role of Klotho Protein in the Development of Cardiovascular Diseases. BioMed Research International. 2018;2018:5171945. doi: 10.1155/2018/5171945Ruiz S, Pergola PE, Zager RA, Vaziri ND. Targeting the transcription factor Nrf2 to ameliorate oxidative stress and inflammation in chronic kidney disease. Kidney International. 2013 Jun;83(6):1029–41. doi: 10.1038/ki.2012.439Ruiz S, Pergola PE, Zager RA, Vaziri ND. Targeting the transcription factor Nrf2 to ameliorate oxidative stress and inflammation in chronic kidney disease. Kidney International. 2013 Jun;83(6):1029–41. doi: 10.1038/ki.2012.439Shen S, Ji C, Wei K. Cellular Senescence and Regulated Cell Death of Tubular Epithelial Cells in Diabetic Kidney Disease. Frontiers in Endocrinology. 2022 Jun 28;13:924299. doi: 10.3389/fendo.2022.924299Shen S, Ji C, Wei K. Cellular Senescence and Regulated Cell Death of Tubular Epithelial Cells in Diabetic Kidney Disease. Frontiers in Endocrinology. 2022 Jun 28;13:924299. doi: 10.3389/fendo.2022.924299Tohyama O, Imura A, Iwano A, Freund JN, Henrissat B, Fujimori T, et al. Klotho Is a Novel βGlucuronidase Capable of Hydrolyzing Steroid βGlucuronides. Journal of Biological Chemistry. 2004 Mar 12;279(11):9777–84. doi: 10.1074/jbc.M312392200Tohyama O, Imura A, Iwano A, Freund JN, Henrissat B, Fujimori T, et al. Klotho Is a Novel βGlucuronidase Capable of Hydrolyzing Steroid βGlucuronides. Journal of Biological Chemistry. 2004 Mar 12;279(11):9777–84. doi: 10.1074/jbc.M312392200Typiak M, Piwkowska A. Antiinflammatory Actions of Klotho: Implications for Therapy of Diabetic Nephropathy. International Journal of Molecular Scienes. 2021 Jan 19;22(2):956–6. doi: 10.3390/ijms22020956Typiak M, Piwkowska A. Antiinflammatory Actions of Klotho: Implications for Therapy of Diabetic Nephropathy. International Journal of Molecular Scienes. 2021 Jan 19;22(2):956–6. doi: 10.3390/ijms22020956Tang A, Zhang Y, Wu L, Lin Y, Lizeyu L, Zhao L, et al. Klotho’s impact on diabetic nephropathy and its emerging connection to diabetic retinopathy. Frontiers in Endocrinology. 2023 Apr 18;14:1180169. doi: 10.3389/fendo.2023.1180169Tang A, Zhang Y, Wu L, Lin Y, Lizeyu L, Zhao L, et al. Klotho’s impact on diabetic nephropathy and its emerging connection to diabetic retinopathy. Frontiers in Endocrinology. 2023 Apr 18;14:1180169. doi: 10.3389/fendo.2023.1180169Casanova A, Wevers A, Navarro-Ledesma S, Pruimboom L. Mitochondria: It is all about energy. Frontiers in Physiology. 2023 Apr 25;14:1114231. doi: 10.3389/fphys.2023.1114231Casanova A, Wevers A, Navarro-Ledesma S, Pruimboom L. Mitochondria: It is all about energy. Frontiers in Physiology. 2023 Apr 25;14:1114231. doi: 10.3389/fphys.2023.1114231

The authors declare that there are no conflicts of interest present.