Исследование уровней экспрессии сплайсинговых изоформ SIRT1 и генов – регуляторов митохондриального гомеостаза в печени больных сахарным диабетом 2-го типа и ожирением

Цель. Оценка ассоциации между уровнем экспрессии изоформ сиртуина 1 (SIRT1) и генами белков, связанных с митохондриальным гомеостазом (PGC-1α, PPAR-γ, PPAR-α, TFAM, MFN2, OPA1, DRP1) в печени больных сахарным диабетом второго типа (СД2).
Материалы и методы. В исследование включено 59 пациентов, которые были разделены на две группы: 1) контрольная группа, индекс массы тела (ИМТ) менее 30 кг/м2, без кардиометаболических нарушений; 2) пациенты с СД2, ИМТ более 30 кг/м2. Выполнялся биохимический анализ показателей крови пациентов, а уровень экспрессии генов интереса в печеночной ткани изучали с помощью количественной полимеразной цепной реакцией с обратной транскрипцией.
Результаты. Обнаружено, что сплайсинговые изоформы SIRT1 V1, V2 и V3 стабильно экспрессировались в печени у больных СД2. Выявлено, что изоформы SIRT1 встречаются не только по отдельности, но и в различных сочетаниях. Экспрессия изоформы SIRT1 V3 значимо повышалась в группе больных, в то время как остальные аналиты значимо не различались между группами. Изоформа SIRT1 V3 положительно коррелировала с уровнем глюкозы. Стоит отметить, что общий SIRT1 не показал значимых корреляций с генами интереса и биохимическими показателями, что только подтверждает необходимость изучения экспрессии изоформ отдельно.
Заключение. Изоформы SIRT1 стабильно экспрессировались в печени, уровень экспрессии изоформы SIRT1 V3 был значимо выше у больных СД2. Результаты работы могут послужить основой для дальнейших, более точечных исследований взаимодействий между сплайсинговыми изоформами SIRT1 с белками митохондриального гомеостаза на посттрансляционном уровне.

1. Tinajero M.G., Malik V.S. An update on the epidemiology of type 2 diabetes: a global perspective. Endocrinol. Metab. Clin. North Am. 2021;50:337–355. DOI: 10.1016/j.ecl.2021.05.013.

2. En Li Cho E., Ang C.Z., Quek J., Fu C.E., Lim L.K.E., Heng Z.E.Q. et al. Global prevalence of non-alcoholic fatty liver disease in type 2 diabetes mellitus: an updated systematic review and meta-analysis. Gut. 2023;72:2138–2148. DOI: 10.1136/gutjnl-2023-330110.

3. Nogueira J.P., Cusi K. Role of insulin resistance in the development of nonalcoholic fatty liver disease in people with type 2 diabetes: from bench to patient care. Diabetes Spectr. 2024;37:20–28. DOI: 10.2337/dsi23-0013.

4. Galicia-Garcia U., Benito-Vicente A., Jebari S., LarreaSebal A., Siddiqi H., Uribe K.B. et al. Pathophysiology of type 2 diabetes mellitus. Int. J. Mol. Sci. 2020;21:6275. DOI: 10.3390/ijms21176275.

5. Chen Z., Tian R., She Z., Cai J., Li H. Role of oxidative stress in the pathogenesis of nonalcoholic fatty liver disease. Free Radic. Biol. Med. 2020;152:116–141. DOI: 10.1016/j.freeradbiomed.2020.02.025.

6. Bhatti J.S., Sehrawat A., Mishra J., Sidhu I.S., Navik U., Khullar N. et al. Oxidative stress in the pathophysiology of type 2 diabetes and related complications: Current therapeutics strategies and future perspectives. Free Radic. Biol. Med. 2022;184:114–134. DOI: 10.1016/j.freeradbiomed.2022.03.019.

7. Li Y.-F., Xie Z.-F., Song Q., Li J.-Y. Mitochondria homeostasis: Biology and involvement in hepatic steatosis to NASH. Acta Pharmacol. Sin. 2022;43:1141–1155. DOI: 10.1038/s41401-022-00864-z.

8. Apostolova N., Vezza T., Muntane J., Rocha M., Víctor V.M. Mitochondrial dysfunction and mitophagy in type 2 diabetes: pathophysiology and therapeutic targets. Antioxid. Redox Signal. 2023;39:278–320. DOI: 10.1089/ars.2022.0016.

9. Van Huynh T., Rethi L., Rethi L., Chen C.-H., Chen Y.-J., Kao Y.-H. The complex interplay between imbalanced mitochondrial dynamics and metabolic disorders in type 2 diabetes. Cells. 2023;12:1223. DOI: 10.3390/cells12091223.

10. Huang C., Chen L., Li J., Ma J., Luo J., Lv Q. et al. Mitochondrial DNA copy number and risk of diabetes mellitus and metabolic syndrome. J. Clin. Endocrinol. Metab. 2023;109:e406–417. DOI: 10.1210/clinem/dgad403.

11. Whitaker R.M., Corum D., Beeson C.C., Schnellmann R.G. Mitochondrial biogenesis as a pharmacological target: a new approach to acute and chronic diseases. Annu. Rev. Pharmacol. Toxicol. 2016;56:229–249. DOI: 10.1146/annurev-pharmtox-010715-103155.

12. Rovira-Llopis S., Bañuls C., Diaz-Morales N., Hernandez-Mijares A., Rocha M., Victor V.M. Mitochondrial dynamics in type 2 diabetes: Pathophysiological implications. Redox. Biol. 2017;11:637–645. DOI: 10.1016/j.redox.2017.01.013.

13. Pinti M.V., Fink G.K., Hathaway Q.A., Durr A.J., Kunovac A., Hollander J.M. Mitochondrial dysfunction in type 2 diabetes mellitus: an organ-based analysis. Am. J. Physiol. Endocrinol. Metab. 2019;316:e268–е285. DOI: 10.1152/ajpendo.00314.2018.

14. Sharma K. Mitochondrial hormesis and diabetic complications. Diabetes. 2015;64:663–672. DOI: 10.2337/db14-0874.

15. Desvergne B., Wahli W. Peroxisome proliferator-activated receptors: nuclear control of metabolism. Endocr. Rev. 1999;20:649–688. DOI: 10.1210/edrv.20.5.0380.

16. Pawlak M., Lefebvre P., Staels B. Molecular mechanism of PPARα action and its impact on lipid metabolism, inflammation and fibrosis in non-alcoholic fatty liver disease. J. Hepatol. 2015;62:720–733. DOI: 10.1016/j.jhep.2014.10.039.

17. Ahmadian M., Suh J.M., Hah N., Liddle C., Atkins A.R., Downes M. et al. PPARγ signaling and metabolism: the good, the bad and the future. Nat. Med. 2013;19:557–566. DOI: 10.1038/nm.3159.

18. Scarpulla R.C. Metabolic control of mitochondrial biogenesis through the PGC-1 family regulatory network. Biochim. Biophys. Acta. 2011;1813:1269–1278. DOI: 10.1016/j.bbamcr.2010.09.019.

19. Halling J.F., Pilegaard H. PGC-1α-mediated regulation of mitochondrial function and physiological implications. Appl. Physiol. Nutr. Metab. 2020;45:927–936. DOI: 10.1139/apnm-2020-0005.

20. Chen W., Zhao H., Li Y. Mitochondrial dynamics in health and disease: mechanisms and potential targets. Sig. Transduct. Target Ther. 2023;8:1–25. DOI: 10.1038/s41392-023-01547-9.

21. Aharoni-Simon M., Hann-Obercyger M., Pen S., Madar Z., Tirosh O. Fatty liver is associated with impaired activity of PPARγ-coactivator 1α (PGC1α) and mitochondrial biogenesis in mice. Lab. Invest. 2011;91:1018–1028. DOI: 10.1038/labinvest.2011.55.

22. Misu H., Takamura T., Matsuzawa N., Shimizu A., Ota T., Sakurai M. Genes involved in oxidative phosphorylation are coordinately upregulated with fasting hyperglycaemia in livers of patients with type 2 diabetes. Diabetologia. 2007;50:268–277. DOI: 10.1007/s00125-006-0489-8.

23. Guclu A., Erdur F.M., Turkmen K. The emerging role of sirtuin 1 in cellular metabolism, diabetes mellitus, diabetic kidney disease and hypertension. Exp. Clin. Endocrinol. Diabetes. 2016;124:131–139. DOI: 10.1055/s-0035-1565067.

24. Kitada M., Koya D. SIRT1 in type 2 diabetes: mechanisms and therapeutic potential. Diabetes Metab. J. 2013;37:315–325. DOI: 10.4093/dmj.2013.37.5.315.

25. Tang B.L. Sirt1 and the mitochondria. Mol. Cells. 2016;39:87–95. DOI: 10.14348/molcells.2016.2318.

26. Khan S.A., Sathyanarayan A., Mashek M.T., Ong K.T., Wollaston-Hayden E.E., Mashek D.G. ATGL-catalyzed lipolysis regulates SIRT1 to control PGC-1α/PPAR-α signaling. Diabetes. 2015;64:418–426. DOI: 10.2337/db14-0325.

27. Nemoto S., Fergusson M.M., Finkel T. SIRT1 functionally interacts with the metabolic regulator and transcriptional coactivator PGC-1{alpha}. J. Biol. Chem. 2005;280:16456–16460. DOI: 10.1074/jbc.M501485200.

28. Purushotham A., Schug T.T., Xu Q., Surapureddi S., Guo X., Li X. Hepatocyte-specific deletion of SIRT1 alters fatty acid metabolism and results in hepatic steatosis and inflammation. Cell Metab. 2009;9:327–338. DOI: 10.1016/j.cmet.2009.02.006.

29. Picard F., Kurtev M., Chung N., Topark-Ngarm A., Senawong T., Machado De Oliveira R. et al. Sirt1 promotes fat mobilization in white adipocytes by repressing PPAR-gamma. Nature. 2004;429:771–776. DOI: 10.1038/nature02583.

30. Han L., Zhou R., Niu J., McNutt M.A., Wang P., Tong T. SIRT1 is regulated by a PPAR{γ}-SIRT1 negative feedback loop associated with senescence. Nucleic Acids Res. 2010;38:7458–7471. DOI: 10.1093/nar/gkq609.

31. Hu Z., Zhang H., Wang Y., Li B., Liu K., Ran J. et al. Exercise activates Sirt1-mediated Drp1 acetylation and inhibits hepatocyte apoptosis to improve nonalcoholic fatty liver disease. Lipids Health Dis. 2023;22:33. DOI: 10.1186/s12944-023-01798-z.

32. Sooyeon L., Go K.L., Kim J.-S. Deacetylation of mitofusin-2 by sirtuin-1: A critical event in cell survival after ischemia. Mol. Cell Oncol. 2016;3:e1087452. DOI: 10.1080/23723556.2015.1087452.

33. Samant S.A., Zhang H.J., Hong Z., Pillai V.B., Sundaresan N.R., Wolfgeher D. et al. SIRT3 deacetylates and activates OPA1 to regulate mitochondrial dynamics during stress. Mol. Cell Biol. 2014;34:807–819. DOI: 10.1128/MCB.01483-13.

34. Patyal P., Ameer F.S., Verma A., Zhang X., Azhar G., Shrivastava J. et al. The role of sirtuin-1 isoforms in regulating mitochondrial function. Curr. Issues Mol. Biol. 2024;46:8835–8851. DOI: 10.3390/cimb46080522.

35. Zhang X., Ameer F.S., Azhar G., Wei J.Y. Alternative splicing increases sirtuin gene family diversity and modulates their subcellular localization and function. Int. J. Mol. Sci. 2021;22:473. DOI: 10.3390/ijms22020473.

36. Patti M.E., Butte A.J., Crunkhorn S., Cusi K., Berria R., Kashyap S. et al. Coordinated reduction of genes of oxidative metabolism in humans with insulin resistance and diabetes: Potential role of PGC1 and NRF1. Proc. Natl. Acad. Sci. USA. 2003;100:8466–8471. DOI: 10.1073/pnas.1032913100.

37. Mootha V.K., Lindgren C.M., Eriksson K.-F., Subramanian A., Sihag S., Lehar J. et al. PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nat. Genet. 2003;34:267–273. DOI: 10.1038/ng1180.

38. Alshawsh M.A., Alsalahi A., Alshehade S.A., Saghir S.A.M., Ahmeda A.F., Al Zarzour R.H. et al. A comparison of the gene expression profiles of non-alcoholic fatty liver disease between animal models of a high-fat diet and methionine-choline-deficient diet. Molecules. 2022;27:858. DOI: 10.3390/molecules27030858.

39. Jiang Y., Chen D., Gong Q., Xu Q., Pan D., Lu F. et al. Elucidation of SIRT-1/PGC-1α-associated mitochondrial dysfunction and autophagy in nonalcoholic fatty liver disease. Lipids. Health Dis. 2021;20:40. DOI: 10.1186/s12944-021-01461-5.

40. Shunkina D., Dakhnevich A., Shunkin E., Khaziakhmatova O., Shupletsova V., Vulf M. et al. gp130 Activates mitochondrial dynamics for hepatocyte survival in a model of steatohepatitis. Biomedicines. 2023;11:396. DOI: 10.3390/biomedicines11020396.

41. Gan K.-X., Wang C., Chen J.-H., Zhu C.-J., Song G.-Y. Mitofusin-2 ameliorates high-fat diet-induced insulin resistance in liver of rats. World J. Gastroenterol. 2013;19:1572–1581. DOI: 10.3748/wjg.v19.i10.1572.

42. Shen J., Zhu B. Integrated analysis of the gene expression profile and DNA methylation profile of obese patients with type 2 diabetes. Mol. Med. Rep. 2018;17:7636–7644. DOI: 10.3892/mmr.2018.8804.

43. Sookoian S., Rosselli M.S., Gemma C., Burgueño A.L., Fernández Gianotti T., Castaño G.O. et al. Epigenetic regulation of insulin resistance in nonalcoholic fatty liver disease: impact of liver methylation of the peroxisome proliferator-activated receptor γ coactivator 1α promoter. Hepatology. 2010;52:1992–2000. DOI: 10.1002/hep.23927.

44. Francque S., Verrijken A., Caron S., Prawitt J., Paumelle R., Derudas B. et al. PPARα gene expression correlates with severity and histological treatment response in patients with non-alcoholic steatohepatitis. J. Hepatol. 2015;63:164–173. DOI: 10.1016/j.jhep.2015.02.019.

45. Westerbacka J., Kolak M., Kiviluoto T., Arkkila P., Sirén J., Hamsten A. et al. Genes involved in fatty acid partitioning and binding, lipolysis, monocyte/macrophage recruitment, and inflammation are overexpressed in the human fatty liver of insulin-resistant subjects. Diabetes. 2007;56:2759–2765. DOI: 10.2337/db07-0156.

46. Deng X.-Q., Chen L.-L., Li N.-X. The expression of SIRT1 in nonalcoholic fatty liver disease induced by high-fat diet in rats. Liver Int. 2007;27:708–715. DOI: 10.1111/j.1478-3231.2007.01497.x.

47. Li Y., Xu S., Giles A., Nakamura K., Lee J.W., Hou X. et al. Hepatic overexpression of SIRT1 in mice attenuates endoplasmic reticulum stress and insulin resistance in the liver. FASEB J. 2011;25:1664–1679. DOI: 10.1096/fj.10-173492.

48. Cao Y., Jiang X., Ma H., Wang Y., Xue P., Liu Y. SIRT1 and insulin resistance. J. Diabetes Complications. 2016;30:178–183. DOI: 10.1016/j.jdiacomp.2015.08.022.

49. Wu T., Liu Y., Fu Y., Liu X., Zhou X. Direct evidence of sirtuin downregulation in the liver of non-alcoholic fatty liver disease patients. Ann. Clin. Lab. Sci. 2014;44:410–418.

50. Castro R.E., Ferreira D.M.S., Afonso M.B., Borralho P.M., Machado M.V., Cortez-Pinto H. et al. MIR-34a/SIRT1/p53 is suppressed by ursodeoxycholic acid in the rat liver and activated by disease severity in human non-alcoholic fatty liver disease. J. Hepatology. 2013;58:119–125. DOI: 10.1016/j.jhep.2012.08.008.