Expression of miR-335 and the genes FASN and SIRT 4 in the visceral adipose tissue the consumption of high-calorie food in experiment

The consumption of high-calorie foods is one of the factors leading to the formation of obesity and related complications. This is based on epigenetic mechanisms, including the regulation of gene expression by microRNAs. The aim of the study is to investigate the effect of high fat diet on the expression of miR-335, as well as the genes FASN and SIRT 4 and the proteins they encode in the visceral adipose tissue of Wistar rats. For 8 weeks, in addition to the standard diet of the vivarium, the rats received animal fats (45 % of daily caloric intake). The relative expression of miR-335 and its target genes FASN and SIRT 4 was determined using real-time PCR, and the protein levels of FASN and SIRT 4 in visceral adipose tissue were measured by enzyme-linked immunosorbent assay (ELISA). It was established that against the background of high-fat diet there is an increase in miR-335 expression and a decrease in activity of lipogenic genes FASN and SIRT 4 in visceral fat tissue of rats, which is accompanied by an increase in blood glucose, development of insulin resistance, and disruption of lipid metabolism. The obtained results indicate the important role of miR-335 in regulating metabolic processes in adipose tissue and the prospect of its use as a therapeutic target for the prevention of complications associated with obesity.

1. Vasyukova O. V., Okorokov P. L., Malievskiy O. A., Neimark A. E., Zorin E. A., Yashkov Y. I., Burmitskaya Yu. V., Kopytina D. A., Bezlepkina O. B., Peterkova V. A. Clinical guidelines “Obesity in children”. Ozhirenie i metabolizm = Obesity and Metabolism, 2024, vol. 21, no. 4, pp. 439–453 (in Russian). https://doi.org/10.14341/omet13194

2. Timasheva Ya. R., Balkhiyarova Zh. R., Kochetova O. V. Current state of the obesity research: genetic aspects, the role of microbiome, and susceptibility to COVID-19. Problemy Endokrinologii = Problems of Endocrinology, 2021, vol. 67, no. 4, pp. 20–35 (in Russian). https://doi.org/10.14341/probl12775

3. Dzhumagaziev A. A., Bezrukova D. A., Shilina N. M., Otto N. Yu., Sosinovskaya E. V., Filipchuk A. V. Genetic and epigenetic risk factors for the development of simple obesity in children: a literature review. Pediatricheskaya farmakologiya = Pediatric Pharmacology, 2024, vol. 21, no. 6, pp. 510–515 (in Russian). https://doi.org/10.15690/pf.v21i6.2828

4. Benavides-Aguilar J. A., Torres-Copado A., Isidoro-Sánchez J., Pathak S., Duttaroy A. K., Banerjee A., Paul S. The regulatory role of microRNAs in obesity and obesity-derived ailments. Genes, 2023, vol. 14, no. 11, art. 2070. https://doi.org/10.3390/genes14112070

5. MacDonald-Ramos K., Martínez-Ibarra A., Monroy A., Miranda-Ríos J., Cerbón M. Effect of dietary fatty acids on microRNA expression related to metabolic disorders and inflammation in human and animal trials. Nutrients, 2021, vol. 13, no. 6, art. 1830. https://doi.org/10.3390/nu13061830

6. Ding W., Sun Y., Han Y., Liu Y., Jin S., Chen J., Gou F. Effects of miR-335 on the proliferation in Yak preadipocytes. Journal of Cytology and Cell Anatomy, 2024, vol. 2, no. 1, pp. 1–12.

7. Zhu L., Chen L., Shi C. M., Xu G. F., Xu L. L., Zhu L. L., Guo X. R., Ni Y., Cui Y., Ji C. MiR-335, an adipogenesisrelated microRNA, is involved in adipose tissue inflammation. Cell Biochemistry and Biophysics, 2014, vol. 68, pp. 283–290. https://doi.org/10.1007/s12013-013-9708-3

8. Nakanishi N., Nakagawa Y., Tokushige N., Aoki N., Matsuzaka T., Ishii K., Yahagi N., Kobayashi K., Yatoh S., Takahashi A., Suzuki H., Urayama O., Yamada N., Shimano H. The up-regulation of microRNA-335 is associated with lipid metabolism in liver and white adipose tissue of genetically obese mice. Biochemical and Biophysical Research Communications, 2009, vol. 385, no. 4, pp. 492–496. https://doi.org/10.1016/j.bbrc.2009.05.058

9. Salunkhe V. A., Ofori J. K., Gandasi N. R., Salö S. A., Hansson S., Andersson M. E., Wendt A., Barg S., Esguerra J. L. S., Eliasson L. MiR-335 overexpression impairs insulin secretion through defective priming of insulin vesicles. Physiological Reports, 2017, vol. 5, no. 21, art. e13493. https://doi.org/10.14814/phy2.13493

10. Kern F., Aparicio-Puerta E., Li Y., Fehlmann T., Kehl T., Wagner V., Ray K., Ludwig N., Lenhof H. P., Meese E., Keller A. miRTargetLink 2.0 – interactive miRNA target gene and target pathway networks. Nucleic Acids Research, 2021, vol. 49, no. 1, pp. W409–W416. https://doi.org/10.1093/nar/gkab297

11. Menendez J. A., Cuyàs E., Encinar J. A., Steen T. V., Verdura S., Llop-Hernández À., López J., Serrano-Hervás E., Osuna S., Martin-Castillo B., Lupu R. Fatty acid synthase (FASN) signalome: A molecular guide for precision oncology. Molecular Oncology, 2024, vol. 18, no. 3, pp. 479–516. https://doi.org/10.1002/1878-0261.13582

12. Min Z., Gao J., Yu Y. The roles of mitochondrial SIRT4 in cellular metabolism. Frontiers in Endocrinology, 2019, vol. 9, art. 783. https://doi.org/10.3389/fendo.2018.00783

13. Diaz-Villaseñor A., Palacios-Gonzalez B., Tovar-Palacio C., Tovar A. R., Torres N. A high fat diet reduces the expression of lipogenic, lipolytic and oxidative genes in white adipose tissue. The effect of the concentration and type of fatty acid is dependent of the dietary protein. FASEB Journal, 2010, vol. 4, no. S1, art. 938.11. https://doi.org/10.1096/fasebj.24.1_supplement.938.11

14. Berndt J., Kovacs P., Ruschke K., Klöting N., Fasshauer M., Schön M. R., Körner A., Stumvoll M., Blüher M. Fatty acid synthase gene expression in human adipose tissue: association with obesity and type 2 diabetes. Diabetologia, 2007, vol. 50, pp. 1472–1480. https://doi.org/10.1007/s00125-007-0689-x

15. Fernandez-Real J. M., Menendez J. A., Moreno-Navarrete J. M., Blüher M., Vazquez-Martin A., Vázquez M. J., Ortega F., Diéguez C., Frühbeck G., Ricart W., Vidal-Puig A. Extracellular fatty acid synthase: a possible surrogate biomarker of insulin resistance. Diabetes, 2010, vol. 59, no. 6, pp. 1506–1511. https://doi.org/10.2337/db09-1756