vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ20.635vavilov-2652Research ArticleФИЗИОЛОГИЧЕСКАЯ ГЕНЕТИКАPHYSIOLOGICAL GENETICSВлияние гонадэктомии и эстрадиола на экспрессию генов сигнального каскада инсулина у самок и самцов мышейEffect of gonadectomy and estradiol on the expression of insulin signaling cascade genes in female and male micehttps://orcid.org/0000-0001-7628-5856ЯковлеваТ. В.IakovlevaT. V.

Новосибирск

Novosibirsk

jakov@bionet.nsc.ruhttps://orcid.org/0000-0001-6137-0492КостинаН. Е.KostinaN. E.

Новосибирск

Novosibirsk

https://orcid.org/0000-0002-6417-9893МакароваЕ. Н.MakarovaE. N.

Новосибирск

Novosibirsk

https://orcid.org/0000-0002-7246-4758БажанН. М.BazhanN. M.

Новосибирск

Novosibirsk

Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наукРоссияInstitute of Cytology and Genetics of Siberian Branch of the Russian Academy of SciencesRussian FederationФедеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук; Новосибирский национальный исследовательский государственный университетРоссияInstitute of Cytology and Genetics of Siberian Branch of the Russian Academy of Sciences; Novosibirsk State UniversityRussian Federation202002072020244427434Copyright © Яковлева Т.В., Костина Н.Е., Макарова Е.Н., Бажан Н.М., 20202020Яковлева Т.В., Костина Н.Е., Макарова Е.Н., Бажан Н.М.Iakovleva T.V., Kostina N.E., Makarova E.N., Bazhan N.M.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/2652

В настоящее время показано положительное влияние эстрадиола на чувствительность к инсулину на уровне целого организма у самок и самцов мышей. При этом чувствительность к инсулину в целом у самок выше, чем у самцов, и самцы демонстрируют большую склонность к развитию метаболических нарушений. Предполагают, что данные половые различия объясняются протективным действием эстрадиола у самок, но не у самцов. Эстрадиол является стероидным гормоном, и его действие обусловлено модуляцией экспрессии генов-мишеней, однако влияние эстрадиола на экспрессию генов, кодирующих трансдукцию сигнала инсулина и транспорт глюкозы в клетку, изучено недостаточно. Целью работы было сравнительное исследование молекулярных механизмов влияния эстрадиола на чувствительность к инсулину у мышей обоих полов. Исследовано влияние гонадэктомии и эстрадиола (1 мкг/животное, три дня) на экспрессию генов сигнального каскада инсулина в мышцах, жировой ткани и печени, а также на экспрессию Fgf21, рецепторов эстрадиола (Esr1/2) и транскрипционного фактора Stat3 в печени у самок и самцов мышей. Ложно оперированные (ЛО) самцы отличались от ЛО самок сниженным уровнем эстрадиола, повышенным уровнем глюкозы и большей резистентностью к инсулину. В печени у ЛО самцов уровни мРНК Irs2, Pik3cd и Esr1/2 были ниже, чем у ЛО самок. У самок гонадэктомия снижала уровень эстрадиола в крови, повышала резистентность к инсулину и уровень глюкозы в крови по сравнению с ЛО самками. Введение эстрадиола гонадэктомированным самкам снижало уровень инсулина в крови и резистентность к инсулину. У самцов гонадэктомия, наоборот, повышала уровень эстрадиола в крови, снижала резистентность к инсулину и уровень инсулина в крови. Введение эстрадиола гонадэктомированным самцам не оказывало влияния на исследованные показатели. Развитие инсулинорезистентности у гонадэктомированных самок было ассоциировано со снижением экспрессии гена Irs2 в печени, а повышение чувствительности к инсулину у гонадэктомированных самцов – с увеличением уровней мРНК Irs2 и Pik3cd в печени. Можно предположить, что повышение уровня эстрадиола в крови активирует экспрессию гена Irs2 в печени независимо от пола животного. Также независимо от пола животного эстрадиол, по-видимому, регулирует транспорт глюкозы в жировой ткани: у самок и самцов повышение уровня эстрадиола в крови было ассоциировано со снижением экспрессии гена Slc2a4 в жировой ткани. Таким образом, эффекты эстрадиола на экспрессию генов инсулинового каскада, по-видимому, не зависят от пола животного, но имеют тканевую специфичность. Поскольку молекулярный механизм влияния эстрадиола на экспрессию генов инсулинового каскада у самок и самцов не различается, причиной половых различий в чувствительности к инсулину и скорости развития метаболических нарушений может быть сниженный, по сравнению с самками, уровень эстрадиола в крови и сниженная экспрессия рецепторов эстрадиола в печени.

A positive effect of estradiol on insulin sensitivity has been shown for females and males. Insulin sensitivity is higher in females than in males, and males show a greater tendency to develop metabolic disorders. It is believed that these sex differences are due to a protective effect of estradiol in females, but not in males. Estradiol is a steroid hormone, and its effect is due to the modulation of target gene expression, but the effect of estradiol on the expression of genes encoding insulin signal transduction and glucose transport has not been sufficiently studied. The aim of the study was to compare the molecular mechanisms of the estradiol influence on insulin sensitivity in mice of both sexes. The effect of gonadectomy and estradiol (1 μg/animal, three days) on the expression of insulin signaling cascade genes in muscle, adipose tissue, and liver, as well as on the expression of Fgf21, estradiol receptors (Esr1/2), and transcription factor Stat3 in the liver in female and male mice was investigated. Estradiol levels were lower and glucose blood levels and insulin resistance were higher in Sham operated (Sham) males compared to Sham females. Irs2, Pik3cd, and Esr1/2 mRNA levels were lower in the liver of Sham males than in Sham females. In females, gonadectomy reduced the level of estradiol in the blood, increased insulin resistance and blood glucose levels compared to Sham females. Administration of estradiol to gonadectomized females decreased blood insulin levels and insulin resistance. In males, gonadectomy, on the contrary, increased the blood estradiol level, decreased blood insulin level and insulin resistance. Estradiol did not affect the parameters studied in males. The development of insulin resistance in gonadectomized females was associated with a decreased expression of the Irs2 gene in the liver. Increased insulin sensitivity in gonadectomized males was associated with increased levels of Irs2 and Pik3cd mRNA in the liver. It can be assumed that increasing the level of estradiol in the blood activates the expression of the Irs2 gene in the liver regardless of animal sex. Also, estradiol seems to regulate the transport of glucose in adipose tissue regardless of animal sex: in females and males, an increase in the blood estradiol level was associated with a decrease in the expression of the Slc2a4 gene in adipose tissue. Thus, the effects of estradiol on the expression of insulin cascade genes do not seem to depend on animal sex, but have tissue specificity. Since the molecular mechanism of estradiol influence on the expression of insulin cascade genes in females and males is the same, the cause of sexual differences in insulin sensitivity and the rate of development of metabolic disorders may be a decrease in the level of estradiol in the blood, as well as a decrease in the expression of estradiol receptors in the liver in males compared to females.

гонадэктомияэстрадиолтестостерончувствительность к инсулинуэкспрессия геновмыши линии C57BL/6Jgonadectomyestradioltestosteroneinsulin sensitivitygene expressionC57BL/6J miceThe study was supported by the Russian Science Foundation, project 17-15-01036. Use of the equipment of the Center for Genetic Resources of Laboratory Animals at ICG SB RAS was supported by the Ministry of Education and Science of Russia (unique project identifier RFMEFI62117X0015).ReferencesAkoum S.E., Lamontagne V., Cloutier I., Tanguay J.F. Nature of fatty acids in high fat diets differentially delineates obesity-linked metabolic syndrome components in male and female C57BL/6J mice. Diabetol. Metab. Syndr. 2011;3:34.Akoum S.E., Lamontagne V., Cloutier I., Tanguay J.F. Nature of fatty acids in high fat diets differentially delineates obesity-linked metabolic syndrome components in male and female C57BL/6J mice. Diabetol. Metab. Syndr. 2011;3:34.Allard C., Bonnet F., Xu B., Coons L., Albarado D., Hill C., Fagherazzi G., Korach K.S., Levin E.R., Lefante J., Morrison C., MauvaisJarvis F. Activation of hepatic estrogen receptor-α increases energy expenditure by stimulating the production of fibroblast growth factor 21 in female mice. Mol. Metab. 2019;22:62-70. DOI 10.1016/j.molmet.2019.02.002.Allard C., Bonnet F., Xu B., Coons L., Albarado D., Hill C., Fagherazzi G., Korach K.S., Levin E.R., Lefante J., Morrison C., MauvaisJarvis F. Activation of hepatic estrogen receptor-α increases energy expenditure by stimulating the production of fibroblast growth factor 21 in female mice. Mol. Metab. 2019;22:62-70. DOI 10.1016/j.molmet.2019.02.002.Bryzgalova G., Lundholm L., Portwood N., Gustafsson J.A., Khan A., Efendic S., Dahlman-Wright K. Mechanisms of antidiabetogenic and body weight-lowering effects of estrogen in high-fat diet-fed mice. Am. J. Physiol. Endocrinol. Metab. 2008;295(4):E904-E912.Bryzgalova G., Lundholm L., Portwood N., Gustafsson J.A., Khan A., Efendic S., Dahlman-Wright K. Mechanisms of antidiabetogenic and body weight-lowering effects of estrogen in high-fat diet-fed mice. Am. J. Physiol. Endocrinol. Metab. 2008;295(4):E904-E912.Faustini-Fustini M., Rochira V., Carani C. Oestrogen deficiency in men: where are we today? Eur. J. Endocrinol. 1999;140(2):111-129.Faustini-Fustini M., Rochira V., Carani C. Oestrogen deficiency in men: where are we today? Eur. J. Endocrinol. 1999;140(2):111-129.Fisher F.M., Estall J.L., Adams A.C., Antonellis P.J., Bina H.A., Flier J.S., Kharitonenkov A., Spiegelman B.M., Maratos-Flier E. Integrated regulation of hepatic metabolism by fibroblast growth factor 21 (FGF21) in vivo. Endocrinology. 2011;152(8):2996-3004. DOI 10.1210/en.2011-0281.Fisher F.M., Estall J.L., Adams A.C., Antonellis P.J., Bina H.A., Flier J.S., Kharitonenkov A., Spiegelman B.M., Maratos-Flier E. Integrated regulation of hepatic metabolism by fibroblast growth factor 21 (FGF21) in vivo. Endocrinology. 2011;152(8):2996-3004. DOI 10.1210/en.2011-0281.Gao H., Bryzgalova G., Hedman E., Khan A., Efendic S., Gustafsson J.A., Dahlman-Wright K. Long-term administration of estradiol decreases expression of hepatic lipogenic genes and improves insulin sensitivity in ob/ob mice: a possible mechanism is through direct regulation of signal transducer and activator of transcription 3. Mol. Endocrinol. 2006;20(6):1287-1299.Gao H., Bryzgalova G., Hedman E., Khan A., Efendic S., Gustafsson J.A., Dahlman-Wright K. Long-term administration of estradiol decreases expression of hepatic lipogenic genes and improves insulin sensitivity in ob/ob mice: a possible mechanism is through direct regulation of signal transducer and activator of transcription 3. Mol. Endocrinol. 2006;20(6):1287-1299.Gong Q., Hu Z., Zhang F., Cui A., Chen X., Jiang H., Gao J., Chen X., Han Y., Liang Q., Ye D., Shi L., Chin Y.E., Wang Y., Xiao H., Guo F., Liu Y., Zang M., Xu A., Li Y. Fibroblast growth factor 21 improves hepatic insulin sensitivity by inhibiting mammalian target of rapamycin complex 1 in mice. Hepatology. 2016;64(2):425-438. DOI 10.1002/hep.28523.Gong Q., Hu Z., Zhang F., Cui A., Chen X., Jiang H., Gao J., Chen X., Han Y., Liang Q., Ye D., Shi L., Chin Y.E., Wang Y., Xiao H., Guo F., Liu Y., Zang M., Xu A., Li Y. Fibroblast growth factor 21 improves hepatic insulin sensitivity by inhibiting mammalian target of rapamycin complex 1 in mice. Hepatology. 2016;64(2):425-438. DOI 10.1002/hep.28523.González C., Alonso A., Grueso N.A., Díaz F., Esteban M.M., Fernández S., Patterson A.M. Effect of treatment with different doses of 17-β-estradiol on insulin receptor substrate-1. JOP. J. Pancreas. 2001;2(4):140-149.González C., Alonso A., Grueso N.A., Díaz F., Esteban M.M., Fernández S., Patterson A.M. Effect of treatment with different doses of 17-β-estradiol on insulin receptor substrate-1. JOP. J. Pancreas. 2001;2(4):140-149.Gorres B.K., Bomhoff G.L., Morris J.K., Geiger P.C. In vivo stimulation of oestrogen receptor α increases insulin-stimulated skeletal muscle glucose uptake. J. Physiol. 2011;589(8):2041-2054. DOI 10.1113/jphysiol.2010.199018.Gorres B.K., Bomhoff G.L., Morris J.K., Geiger P.C. In vivo stimulation of oestrogen receptor α increases insulin-stimulated skeletal muscle glucose uptake. J. Physiol. 2011;589(8):2041-2054. DOI 10.1113/jphysiol.2010.199018.Iakovleva T.V., Makarova E.N., Bazhan N.M. Effect of ovariectomy on GLUT4 mRNA levels in adipose and muscle tissues in females of mice C57BL/6J – Agouti yellow. Rossiyskiy Fiziologicheskiy Zhurnal im. I.M. Sechenova = Russian Journal of Physiology. 2014; 100(5):602-612. (in Russian)Iakovleva T.V., Makarova E.N., Bazhan N.M. Effect of ovariectomy on GLUT4 mRNA levels in adipose and muscle tissues in females of mice C57BL/6J – Agouti yellow. Rossiyskiy Fiziologicheskiy Zhurnal im. I.M. Sechenova = Russian Journal of Physiology. 2014; 100(5):602-612. (in Russian)Kim J.Y., Jo K.J., Kim O.S., Kim B.J., Kang D.W., Lee K.H., Baik H.W., Han M.S., Lee S.K. Parenteral 17beta-estradiol decreases fasting blood glucose levels in non-obese mice with short-term ovariectomy. Life Sci. 2010;87(11-12):358-366. DOI 10.1016/j.lfs.2010.07.009.Kim J.Y., Jo K.J., Kim O.S., Kim B.J., Kang D.W., Lee K.H., Baik H.W., Han M.S., Lee S.K. Parenteral 17beta-estradiol decreases fasting blood glucose levels in non-obese mice with short-term ovariectomy. Life Sci. 2010;87(11-12):358-366. DOI 10.1016/j.lfs.2010.07.009.Lindberg M.K., Movérare S., Skrtic S., Gao H., Dahlman-Wright K., Gustafsson J.A., Ohlsson C. Estrogen receptor (ER)-β reduces ERαregulated gene transcription, supporting a “Ying Yang” relationship between ERα and ERβ in mice. Mol. Endocrinol. 2003;17(2):203-208. DOI 10.1210/me.2002-0206.Lindberg M.K., Movérare S., Skrtic S., Gao H., Dahlman-Wright K., Gustafsson J.A., Ohlsson C. Estrogen receptor (ER)-β reduces ERαregulated gene transcription, supporting a “Ying Yang” relationship between ERα and ERβ in mice. Mol. Endocrinol. 2003;17(2):203-208. DOI 10.1210/me.2002-0206.Muthusamy T., Murugesan P., Balasubramanian K. Sex steroids deficiency impairs glucose transporter 4 expression and its translocation through defective Akt phosphorylation in target tissues of adult male rat. Metabolism. 2009;58(11):1581-1592.Muthusamy T., Murugesan P., Balasubramanian K. Sex steroids deficiency impairs glucose transporter 4 expression and its translocation through defective Akt phosphorylation in target tissues of adult male rat. Metabolism. 2009;58(11):1581-1592.Muthusamy T., Murugesan P., Balasubramanian K. Sex steroids influence glucose oxidation through modulation of insulin receptor expression and IRS-1 serine phosphorylation in target tissues of adult male rat. Mol. Cell. Biochem. 2011;352(1-2):35-45.Muthusamy T., Murugesan P., Balasubramanian K. Sex steroids influence glucose oxidation through modulation of insulin receptor expression and IRS-1 serine phosphorylation in target tissues of adult male rat. Mol. Cell. Biochem. 2011;352(1-2):35-45.Narasimhan A., Sampath S., Jayaraman S., Karundevi B. Estradiol favors glucose oxidation in gastrocnemius muscle through modulation of insulin signaling molecules in adult female rats. Endocr. Res. 2013;38(4):251-262. DOI 10.3409/07435800.2013.775148.Narasimhan A., Sampath S., Jayaraman S., Karundevi B. Estradiol favors glucose oxidation in gastrocnemius muscle through modulation of insulin signaling molecules in adult female rats. Endocr. Res. 2013;38(4):251-262. DOI 10.3409/07435800.2013.775148.Oh Y.S., Lee T.S., Cheon G.J., Jang I.S., Jun H.S., Park S.C. Modulation of insulin sensitivity and caveolin-1 expression by orchidectomy in a nonobese type 2 diabetes animal model. Mol. Med. 2011;17(1-2): 4-11. DOI 10.2119/molmed.2009.00105.Oh Y.S., Lee T.S., Cheon G.J., Jang I.S., Jun H.S., Park S.C. Modulation of insulin sensitivity and caveolin-1 expression by orchidectomy in a nonobese type 2 diabetes animal model. Mol. Med. 2011;17(1-2): 4-11. DOI 10.2119/molmed.2009.00105.Panno M.L., Mauro L., Marsico S., Bellizzi D., Rizza P., Morelli C., Salerno M., Giordano F., Andò S. Evidence that the mouse insulin receptor substrate-1 belongs to the gene family on which the promoter is activated by estrogen receptor α through its interaction with Sp1. J. Mol. Endocrinol. 2006;36(1):91-105. DOI 10.1677/jme.1.01848.Panno M.L., Mauro L., Marsico S., Bellizzi D., Rizza P., Morelli C., Salerno M., Giordano F., Andò S. Evidence that the mouse insulin receptor substrate-1 belongs to the gene family on which the promoter is activated by estrogen receptor α through its interaction with Sp1. J. Mol. Endocrinol. 2006;36(1):91-105. DOI 10.1677/jme.1.01848.Parks B.W., Sallam T., Mehrabian M., Psychogios N., Hui S.T., Norheim F., Castellani L.W., Rau C.D., Pan C., Phun J., Zhou Z., Yang W.P., Neuhaus I., Gargalovic P.S., Kirchgessner T.G., Graham M., Lee R., Tontonoz P., Gerszten R.E., Hevener A.L., Lusis A.J. Genetic architecture of insulin resistance in the mouse. Cell Metab. 2015;21(2):334-347.Parks B.W., Sallam T., Mehrabian M., Psychogios N., Hui S.T., Norheim F., Castellani L.W., Rau C.D., Pan C., Phun J., Zhou Z., Yang W.P., Neuhaus I., Gargalovic P.S., Kirchgessner T.G., Graham M., Lee R., Tontonoz P., Gerszten R.E., Hevener A.L., Lusis A.J. Genetic architecture of insulin resistance in the mouse. Cell Metab. 2015;21(2):334-347.Parthasarathy C., Renuka V.N., Balasubramanian K. Sex steroids enhance insulin receptors and glucose oxidation in Chang liver cells. Clin. Chim. Acta. 2009;399(1-2):49-53.Parthasarathy C., Renuka V.N., Balasubramanian K. Sex steroids enhance insulin receptors and glucose oxidation in Chang liver cells. Clin. Chim. Acta. 2009;399(1-2):49-53.Rogers N.H., Perfield J.W. 2nd, Strissel K.J., Obin M.S., Greenberg A.S. Reduced energy expenditure and increased inflammation are early events in the development of ovariectomy-induced obesity. Endocrinology. 2009;150(5):2161-2168. DOI 10.1210/en.2008-1405.Rogers N.H., Perfield J.W. 2nd, Strissel K.J., Obin M.S., Greenberg A.S. Reduced energy expenditure and increased inflammation are early events in the development of ovariectomy-induced obesity. Endocrinology. 2009;150(5):2161-2168. DOI 10.1210/en.2008-1405.Saengsirisuwan V., Pongseeda S., Prasannarong M., Vichaiwong K., Toskulkao C. Modulation of insulin resistance in ovariectomized rats by endurance exercise training and estrogen replacement. Metabolism. 2009;58(1):38-47. DOI 10.1016/j.metabol.2008.08.004.Saengsirisuwan V., Pongseeda S., Prasannarong M., Vichaiwong K., Toskulkao C. Modulation of insulin resistance in ovariectomized rats by endurance exercise training and estrogen replacement. Metabolism. 2009;58(1):38-47. DOI 10.1016/j.metabol.2008.08.004.Torre S.D., Lolli F., Ciana P., Maggi A. Sexual dimorphism and estrogen action in mouse liver. In: Mauvais-Jarvis F. (Ed.). Sex and Gender Factors Affecting Metabolic Homeostasis, Diabetes and Obesity (Advances in Experimental Medicine and Biology. Vol. 1043). Springer Int. Publ., 2017;141-151. DOI 10.1007/978-3-319-70178-3_8.Torre S.D., Lolli F., Ciana P., Maggi A. Sexual dimorphism and estrogen action in mouse liver. In: Mauvais-Jarvis F. (Ed.). Sex and Gender Factors Affecting Metabolic Homeostasis, Diabetes and Obesity (Advances in Experimental Medicine and Biology. Vol. 1043). Springer Int. Publ., 2017;141-151. DOI 10.1007/978-3-319-70178-3_8.Xie P., Liu M.L., Gu Y.P., Lu J., Xu X., Zeng W.M., Song H.P. Oestrogen improves glucose metabolism and insulin signal transduction in HepG2 cells. Clin. Exp. Pharmacol. Physiol. 2003;30(9):643-648. DOI 10.1046/j.1440-1681.2003.03899.x.Xie P., Liu M.L., Gu Y.P., Lu J., Xu X., Zeng W.M., Song H.P. Oestrogen improves glucose metabolism and insulin signal transduction in HepG2 cells. Clin. Exp. Pharmacol. Physiol. 2003;30(9):643-648. DOI 10.1046/j.1440-1681.2003.03899.x.Yakovleva T.V., Kazantseva A.Yu., Makarova E.N., Bazhan N.M. Sex differences of molecular mechanisms of insulin sensitivity in young and adult C57BL/6J mice. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2017;21(7):833-840. DOI 10.18699/VJ17.303. (in Russian)Yakovleva T.V., Kazantseva A.Yu., Makarova E.N., Bazhan N.M. Sex differences of molecular mechanisms of insulin sensitivity in young and adult C57BL/6J mice. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2017;21(7):833-840. DOI 10.18699/VJ17.303. (in Russian)Zhu L., Martinez M.N., Emfinger C.H., Palmisano B.T., Stafford J.M. Estrogen signaling prevents diet-induced hepatic insulin resistance in male mice with obesity. Am. J. Physiol. Endocrinol. Metab. 2014; 306(10):E1188-E1197. DOI 10.1152/ajpendo.00579.2013.Zhu L., Martinez M.N., Emfinger C.H., Palmisano B.T., Stafford J.M. Estrogen signaling prevents diet-induced hepatic insulin resistance in male mice with obesity. Am. J. Physiol. Endocrinol. Metab. 2014; 306(10):E1188-E1197. DOI 10.1152/ajpendo.00579.2013.

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