Effects of progesterone on the lipolysis of lipid droplets and prostaglandin E2 synthesis in murine cervical epithelial cells
Hongyan Zhang A * , Feng Su A * , Libo Huang A * , Boyu Li A , Xuejun Yuan B , Mingjiu Luo A C and Lijiang Ge A CA College of Animal Science and Technology, No. 61 Daizong Street, Taian, Shandong Province, 271018, PR China.
B College of Life Science, Shandong Agricultural University, No. 61 Daizong Street, Taian, Shandong Province, 271018, PR China.
C Corresponding authors. Email: glj@sdau.edu.cn; luo9616@163.com
Reproduction, Fertility and Development - https://doi.org/10.1071/RD20195
Submitted: 26 July 2020 Accepted: 13 November 2020 Published online: 28 January 2021
Abstract
Previous studies demonstrated that progesterone (P4) can promote prostaglandin (PG) E2 production; however, how P4 mediates the synthesis of PGE2 remains unclear. In this study, cervical epithelial cells from mice during the follicular phase were cultured in vitro and treated with different concentrations of P4 (5, 10, and 20 nM). The results of the present study suggest that treatment of murine cervical epithelial cells with 10 nM P4 for 24 h contributed to: (1) significantly increased expression of protein kinase A (PKA), cytosolic phospholipase A2 (cPLA2) and PGE synthase (PGES)-1; (2) higher phosphorylated (p-) to total extracellular signal-regulated kinase (ERK) 1/2 and hormone-sensitive lipase (HSL) ratios; (3) a significant decrease in the number of lipid droplets (LDs) and fatty acid content within LDs in epithelial cells; and (4) enhanced arachidonic acid and PGE2 levels in cells compared with the control (0 nM P4) group (P < 0.01 for all findings). In contrast, the PKA inhibitor H89 contributed to significantly decreased cPLA2, PGES-1 and HSL expression, ERK1/2 phosphorylation and arachidonic acid and PGE2 levels, even in the presence of P4. These data show that P4 can act via the PKA/ERK1/2 pathway to stimulate lipolysis of triacylglycerol in the LD core and degradation of phospholipid in the LD membrane to promote PGE2 synthesis in murine cervical epithelial cells.
Keywords: cervical epithelial cells, lipid droplets, neutral lipase, progesterone, prostaglandin, protein kinase A/extracellular signal-regulated kinase 1/2 pathway.
References
Accioly, M. T., Pacheco, P., Maya-Monteiro, C. M., Carrossini, N., Robbs, B. K., Oliveira, S. S., Kaufmann, C., Morgado-Diaz, J. A., Bozza, P. T., and Viola, J. P. (2008). Lipid bodies are reservoirs of cyclooxygenase-2 and sites of prostaglandin-E2 synthesis in colon cancer cells. Cancer Res. 68, 1732–1740.| Lipid bodies are reservoirs of cyclooxygenase-2 and sites of prostaglandin-E2 synthesis in colon cancer cells.Crossref | GoogleScholarGoogle Scholar | 18339853PubMed |
Anfuso, C. D., Lupo, G., Romeo, L., Giurdanella, G., Motta, C., Pascale, A., Tirolo, C., Marchetti, B., and Alberghina, M. (2007). Endothelial cell–pericyte co-cultures induce PLA2 protein expression through activation of PKCa and the MAPK/ERK cascade. J. Lipid Res. 48, 782–793.
| Endothelial cell–pericyte co-cultures induce PLA2 protein expression through activation of PKCa and the MAPK/ERK cascade.Crossref | GoogleScholarGoogle Scholar | 17267947PubMed |
Asselin, E., Goff, A. K., Bergeron, H., and Fortier, M. A. (1996). Influence of sex steroids on the production of prostaglandins F2α and E2 and response to oxytocin in cultured epithelial and stromal cells of the bovine endometrium. Biol. Reprod. 54, 371–379.
| Influence of sex steroids on the production of prostaglandins F2α and E2 and response to oxytocin in cultured epithelial and stromal cells of the bovine endometrium.Crossref | GoogleScholarGoogle Scholar | 8788188PubMed |
Bódis, K., and Roden, M. (2018). Energy metabolism of white adipose tissue and insulin resistance in humans. Eur. J. Clin. Invest. 48, e13017.
| Energy metabolism of white adipose tissue and insulin resistance in humans.Crossref | GoogleScholarGoogle Scholar | 30107041PubMed |
Bolte, S., and Cordelieres, F. P. (2006). A guided tour into subcellular colocalization analysis in light microscopy. J. Microsc. 224, 213–232.
| 17210054PubMed |
Bozza, P. T., and Weller, P. F. (2001). Arachidonyl trifluoromethyl ketone induces lipid body formation in leukocytes. Prostaglandins Leukot. Essent. Fatty Acids 64, 227–230.
| Arachidonyl trifluoromethyl ketone induces lipid body formation in leukocytes.Crossref | GoogleScholarGoogle Scholar | 11418016PubMed |
Bozza, P. T., Yu, W., Penrose, J. F., Morgan, E. S., Dvorak, A. M., and Weller, P. F. (1997). Eosinophil lipid bodies: specific, inducible intracellular sites for enhanced eicosanoid formation. J. Exp. Med. 186, 909–920.
| Eosinophil lipid bodies: specific, inducible intracellular sites for enhanced eicosanoid formation.Crossref | GoogleScholarGoogle Scholar | 9294145PubMed |
Bozza, P. T., Bakker-Abreu, I., Navarro-Xavier, R. A., and Bandeira-Melo, C. (2011). Lipid body function in eicosanoid synthesis: an update. Prostaglandins Leukot. Essent. Fatty Acids 85, 205–213.
| Lipid body function in eicosanoid synthesis: an update.Crossref | GoogleScholarGoogle Scholar | 21565480PubMed |
Brinsfield, T. H., and Hawk, H. W. (1973). Control by progesterone of the concentration of lipid droplets in epithelial cells of the sheep endometrium. J. Anim. Sci. 36, 919–922.
| Control by progesterone of the concentration of lipid droplets in epithelial cells of the sheep endometrium.Crossref | GoogleScholarGoogle Scholar | 4735755PubMed |
Bush, K. T., Lee, H., and Nagele, R. G. (1992). Lipid droplets of neuroepithelial cells are a major calcium storage site during neural tube formation in chick and mouse embryos. Experientia 48, 516–519.
| Lipid droplets of neuroepithelial cells are a major calcium storage site during neural tube formation in chick and mouse embryos.Crossref | GoogleScholarGoogle Scholar | 1601118PubMed |
Byers, S. L., Wiles, M. V., Dunn, S. L., and Taft, R. A. (2012). Mouse estrous cycle identification tool and images. PLoS One 7, e35538.
| Mouse estrous cycle identification tool and images.Crossref | GoogleScholarGoogle Scholar | 22514749PubMed |
Chang, L., and Karin, M. (2001). Mammalian MAP kinase signalling cascades. Nature 410, 37–40.
| Mammalian MAP kinase signalling cascades.Crossref | GoogleScholarGoogle Scholar | 11242034PubMed |
Clifford, G. M., Londos, C., Kraemer, F. B., Vernon, R. G., and Yeaman, S. J. (2000). Translocation of hormone-sensitive lipase and perilipin upon lipolytic stimulation of rat adipocytes. J. Biol. Chem. 275, 5011–5015.
| Translocation of hormone-sensitive lipase and perilipin upon lipolytic stimulation of rat adipocytes.Crossref | GoogleScholarGoogle Scholar | 10671541PubMed |
Cohen, B. C., Raz, C., Shamay, A., and Argov-Argaman, N. (2017). Lipid droplet fusion in mammary epithelial cells is regulated by phosphatidylethanolamine metabolism. J. Mammary Gland Biol. Neoplasia 22, 235–249.
| Lipid droplet fusion in mammary epithelial cells is regulated by phosphatidylethanolamine metabolism.Crossref | GoogleScholarGoogle Scholar | 29188493PubMed |
Cook, H. W., Clarke, J. T. R., and Spence, M. W. (1982). Involvement of triacylglycerol in the metabolism of fatty acids by cultured neuroblastoma and glioma cell. J. Lipid Res. 23, 1292–1300.
| 7161559PubMed |
Duras, M., Mlynarczuk, J., and Kotwica, J. (2005). Non-genomic effect of steroids on oxytocin-stimulated intracellular mobilisation of calcium and on prostaglandin F2α and E2 secretion from bovine endometrial cells. Prostaglandins Other Lipid Mediat. 76, 105–116.
| Non-genomic effect of steroids on oxytocin-stimulated intracellular mobilisation of calcium and on prostaglandin F2α and E2 secretion from bovine endometrial cells.Crossref | GoogleScholarGoogle Scholar | 15967166PubMed |
Fan, T., Li, X., Li, Y., Zhi, Y., Rong, S., Cheng, G., and Zhang, X. (2018). An improved method for primary culture of normal cervical epithelial cells and establishment of cell model in vitro with HPV-16 E6 gene by lentivirus. J Cell Physiol. 233, 2773–2780.
| An improved method for primary culture of normal cervical epithelial cells and establishment of cell model in vitro with HPV-16 E6 gene by lentivirus.Crossref | GoogleScholarGoogle Scholar | 28464265PubMed |
Farese, R. V., and Walther, T. C. (2009). Lipid droplets finally get a little R-E-S-P-E-C-T. Cell 139, 855–860.
| Lipid droplets finally get a little R-E-S-P-E-C-T.Crossref | GoogleScholarGoogle Scholar | 19945371PubMed |
Fujinoki, M. (2013). Progesterone-enhanced sperm hyperactivation through IP3–PKC and PKA signals. Reprod. Med. Biol. 12, 27–33.
| Progesterone-enhanced sperm hyperactivation through IP3–PKC and PKA signals.Crossref | GoogleScholarGoogle Scholar | 29699127PubMed |
Garg, D., Ng, S. S. M., Baig, K. M., Driggers, P., and Segars, J. (2017). Progesterone-mediated non-classical signaling. Trends Endocrinol. Metab. 28, 656–668.
| Progesterone-mediated non-classical signaling.Crossref | GoogleScholarGoogle Scholar | 28651856PubMed |
Gu, G., Gao, Q., Yuan, X., Huang, L., and Ge, L. (2012). Immunolocalization of adipocytes and prostaglandin E2 and Its four receptor proteins EP1, EP2, EP3, and EP4 in the caprine cervix during spontaneous term labor. Biol. Reprod. 86, 159.
| Immunolocalization of adipocytes and prostaglandin E2 and Its four receptor proteins EP1, EP2, EP3, and EP4 in the caprine cervix during spontaneous term labor.Crossref | GoogleScholarGoogle Scholar | 22402965PubMed |
Jacobsen, B. M., and Horwitz, K. B. (2012). Progesterone receptors, their isoforms and progesterone regulated transcription. Mol. Cell. Endocrinol. 357, 18–29.
| Progesterone receptors, their isoforms and progesterone regulated transcription.Crossref | GoogleScholarGoogle Scholar | 21952082PubMed |
Karteris, E., Zervou, S., Pang, Y., Dong, J., Hillhouse, E. W., Randeva, H. S., and Thomas, P. (2006). Progesterone signalling in human myometrium through two novel membrane G protein-coupled receptors: potential role in functional progesterone withdrawal at term. Mol. Endocrinol. 20, 1519–1534.
| Progesterone signalling in human myometrium through two novel membrane G protein-coupled receptors: potential role in functional progesterone withdrawal at term.Crossref | GoogleScholarGoogle Scholar | 16484338PubMed |
Kimmel, A. R., Brasaemle, D. L., McAndrews-Hill, M., Sztalryd, C., and Londos, C. (2010). Adoption of PERILIPIN as a unifying nomenclature for the mammalian PAT-family of intracellular lipid storage droplet proteins. J. Lipid Res. 51, 468–471.
| Adoption of PERILIPIN as a unifying nomenclature for the mammalian PAT-family of intracellular lipid storage droplet proteins.Crossref | GoogleScholarGoogle Scholar | 19638644PubMed |
Kuse, M., Sakumoto, R., and Okuda, K. (2016). Genomic and non-genomic effects of progesterone on prostaglandin (PG) F2α and PGE2 production in the bovine endometrium. Reprod. Fertil. Dev. 28, 1588–1597.
| Genomic and non-genomic effects of progesterone on prostaglandin (PG) F2α and PGE2 production in the bovine endometrium.Crossref | GoogleScholarGoogle Scholar |
Lange, C. A. (2004). Making sense of cross-talk between steroid hormone receptors and intracellular signalling pathways: who will have the last word? Mol. Endocrinol. 18, 269–278.
| Making sense of cross-talk between steroid hormone receptors and intracellular signalling pathways: who will have the last word?Crossref | GoogleScholarGoogle Scholar | 14563938PubMed |
Leonhardt, S. A., and Edwards, D. P. (2002). Mechanism of action of progesterone antagonists. Exp. Biol. Med. (Maywood) 227, 969–980.
| Mechanism of action of progesterone antagonists.Crossref | GoogleScholarGoogle Scholar | 12486206PubMed |
Mani, S. K., Mermelstein, P. G., Tetel, M. J., and Anesetti, G. (2012). Convergence of multiple mechanisms of steroid hormone action. Horm. Metab. Res. 44, 569–576.
| Convergence of multiple mechanisms of steroid hormone action.Crossref | GoogleScholarGoogle Scholar | 22454239PubMed |
Martin, S., and Parton, R. G. (2006). Lipid droplets: a unified view of a dynamic organelle. Nat. Rev. Mol. Cell Biol. 7, 373–378.
| Lipid droplets: a unified view of a dynamic organelle.Crossref | GoogleScholarGoogle Scholar | 16550215PubMed |
Moreira, L. S., Piva, B., Gentile, L. B., Mesquita-Santos, F. P., D’Avila, H., Maya-Monteiro, C. M., Bozza, P. T., Bandeira-Melo, C., and Diaz, B. L. (2009). Cytosolic phospholipase A2-driven PGE2 synthesis within unsaturated fatty acids-induced lipid bodies of epithelial cells. Biochim. Biophys. Acta 1791, 156–165.
| Cytosolic phospholipase A2-driven PGE2 synthesis within unsaturated fatty acids-induced lipid bodies of epithelial cells.Crossref | GoogleScholarGoogle Scholar | 19367763PubMed |
Morel, E., Ghezzal, S., Lucchi, G., Truntzer, C., Pais de Barros, J. P., Simon-Plas, F., Demignot, S., Mineo, C., Shaul, P. W., Leturque, A., Rousset, M., and Carrière, V. (2018). Cholesterol trafficking and raft-like membrane domain composition mediate scavenger receptor class B type 1-dependent lipid sensing in intestinal epithelial cells. Biochim. Biophys. Acta 1863, 199–211.
| Cholesterol trafficking and raft-like membrane domain composition mediate scavenger receptor class B type 1-dependent lipid sensing in intestinal epithelial cells.Crossref | GoogleScholarGoogle Scholar |
Mueck, A. O., Ruan, X., Seeger, H., Fehm, T., and Neubauer, H. (2014). Genomic and non-genomic actions of progestogens in the breast. J. Steroid Biochem. Mol. Biol. 142, 62–67.
| Genomic and non-genomic actions of progestogens in the breast.Crossref | GoogleScholarGoogle Scholar | 23994274PubMed |
Neulen, J., Zahradnik, H. P., Flecken, U., and Breckwoldt, M. (1988). Effects of estradiol-17 beta and progesterone on the synthesis of prostaglandin F2 alpha, prostaglandin E2 and prostaglandin I2 by fibroblasts from human endometrium in vitro. Prostaglandins 36, 17–30.
| Effects of estradiol-17 beta and progesterone on the synthesis of prostaglandin F2 alpha, prostaglandin E2 and prostaglandin I2 by fibroblasts from human endometrium in vitro.Crossref | GoogleScholarGoogle Scholar | 3051134PubMed |
Plewes, M. R., and Burns, P. D. (2018). Effect of fish oil on agonist-induced receptor internalization of the PGF2α receptor and cell signaling in bovine luteal cells in vitro. Domest. Anim. Endocrinol. 63, 38–47.
| Effect of fish oil on agonist-induced receptor internalization of the PGF2α receptor and cell signaling in bovine luteal cells in vitro.Crossref | GoogleScholarGoogle Scholar | 29306078PubMed |
Qiu, B., and Simon, M. C. (2016). BODIPY 493/503 staining of neutral lipid droplets for microscopy and quantification by flow cytometry. Bio Protoc. 6, e1912.
| BODIPY 493/503 staining of neutral lipid droplets for microscopy and quantification by flow cytometry.Crossref | GoogleScholarGoogle Scholar | 28573161PubMed |
Roskoski, R. (2012). ERK1/2 MAP kinases: structure, function, and regulation. Pharmacol. Res. 66, 105–143.
| ERK1/2 MAP kinases: structure, function, and regulation.Crossref | GoogleScholarGoogle Scholar | 22569528PubMed |
Schaloske, R. H., and Dennis, E. A. (2006). The phospholipase A2 superfamily and its group numbering system. Biochim. Biophys. Acta 1761, 1246–1259.
| The phospholipase A2 superfamily and its group numbering system.Crossref | GoogleScholarGoogle Scholar | 16973413PubMed |
Stelmanska, E., Szrok, S., and Swierczynski, J. (2015). Progesterone-induced down-regulation of hormone sensitive lipase (Lipe) and up-regulation of G0/G1 switch 2 (G0s2) genes expression in inguinal adipose tissue of female rats is reflected by diminished rate of lipolysis. J. Steroid Biochem. Mol. Biol. 147, 31–39.
| Progesterone-induced down-regulation of hormone sensitive lipase (Lipe) and up-regulation of G0/G1 switch 2 (G0s2) genes expression in inguinal adipose tissue of female rats is reflected by diminished rate of lipolysis.Crossref | GoogleScholarGoogle Scholar | 25448749PubMed |
Sztalryd, C., Xu, G., Dorward, H., Tansey, J. T., Contreras, J. A., Kimmel, A. R., and Londos, C. (2003). Perilipin A is essential for the translocation of hormone-sensitive lipase during lipolytic activation. J. Cell Biol. 161, 1093–1103.
| Perilipin A is essential for the translocation of hormone-sensitive lipase during lipolytic activation.Crossref | GoogleScholarGoogle Scholar | 12810697PubMed |
Tamura, K., Naraba, H., Hara, T., Nakamura, K., Yoshie, M., Kogo, H., and Tachikawa, E. (2016). A positive feedback loop between progesterone and microsomal prostaglandin E synthase-1-mediated PGE2 promotes production of both in mouse granulosa cells. Prostaglandins Other Lipid Mediat. 123, 56–62.
| A positive feedback loop between progesterone and microsomal prostaglandin E synthase-1-mediated PGE2 promotes production of both in mouse granulosa cells.Crossref | GoogleScholarGoogle Scholar | 27174800PubMed |
Tao, L., Zhang, H., Wang, H., Li, L., Huang, L., Su, F., Yuan, X., Luo, M., and Ge, L. (2020). Characteristics of lipid droplets and the expression of proteins involved in lipolysis in the murine cervix during mid-pregnancy. Reprod. Fertil. Dev. 32, 967–975.
| Characteristics of lipid droplets and the expression of proteins involved in lipolysis in the murine cervix during mid-pregnancy.Crossref | GoogleScholarGoogle Scholar | 32693909PubMed |
Tauchi-Sato, K., Ozeki, S., Houjou, T., Taguchi, R., and Fujimoto, T. (2002). The surface of lipid droplets is a phospholipid monolayer with a unique fatty acid composition. J. Biol. Chem. 277, 44507–44512.
| The surface of lipid droplets is a phospholipid monolayer with a unique fatty acid composition.Crossref | GoogleScholarGoogle Scholar | 12221100PubMed |
Triggiani, M., Seeds, M. C., Bass, D. A., Marone, G., and Chilton, F. H. (1995). Migration of human inflammatory cells into the lung results in the remodeling of arachidonic acid into a triglyceride pool. J. Exp. Med. 182, 1181–1190.
| Migration of human inflammatory cells into the lung results in the remodeling of arachidonic acid into a triglyceride pool.Crossref | GoogleScholarGoogle Scholar | 7595189PubMed |
Vicent, G. P., Nacht, A. S., Zaurin, R., Font-Mateu, J., Soronellas, D., Le Dily, F., Reyes, D., and Beato, M. (2013). Unliganded progesterone receptor-mediated targeting of an RNA-containing repressive complex silences a subset of hormone-inducible genes. Genes Dev. 27, 1179–1197.
| Unliganded progesterone receptor-mediated targeting of an RNA-containing repressive complex silences a subset of hormone-inducible genes.Crossref | GoogleScholarGoogle Scholar | 23699411PubMed |
Vincent, D. L., Meredith, S., and Inskeep, E. K. (1986). Advancement of uterine secretion of prostaglandin E2 by treatment with progesterone and transfer of asynchronous embryos. Endocrinology 119, 527–529.
| Advancement of uterine secretion of prostaglandin E2 by treatment with progesterone and transfer of asynchronous embryos.Crossref | GoogleScholarGoogle Scholar | 3460797PubMed |
Weller, P. F., Monahan-Earley, R. A., Dvorak, H. F., and Dvorak, A. M. (1991). Cytoplasmic lipid bodies of human eosinophils: subcellular isolation and analysis of arachidonate incorporation. Am. J. Pathol. 138, 141–148.
| 1846262PubMed |
Yagel, S., Hurwitz, A., Rosenn, B., and Keizer, N. (1987). Progesterone enhancement of prostaglandin E2 production by fetal placental macrophages. Am. J. Reprod. Immunol. Microbiol. 14, 45–48.
| Progesterone enhancement of prostaglandin E2 production by fetal placental macrophages.Crossref | GoogleScholarGoogle Scholar | 3475985PubMed |
Yoon, S., and Seger, R. (2006). The extracellular signal-regulated kinase: multiple substrates regulate diverse cellular functions. Growth Factors 24, 21–44.
| The extracellular signal-regulated kinase: multiple substrates regulate diverse cellular functions.Crossref | GoogleScholarGoogle Scholar | 16393692PubMed |
Yu, W., Bozza, P. T., Tzizik, D. M., Gray, J. P., Cassara, J., Cvorak, A. M., and Weller, P. F. (1998). Co-compartmentalization of MAP kinases and cytosolic phospholipase A2 at cytoplasmic arachidonate-rich lipid bodies. Am. J. Pathol. 152, 759–769.
| 9502418PubMed |
Zhu, Y., Rice, C. D., Pang, Y., Pace, M., and Thomas, P. (2003). Cloning, expression and characterisation of a membrane progestin receptor and evidence it is an intermediary in meiotic maturation of fish oocytes. Proc. Natl Acad. Sci. USA 100, 2231–2236.
| Cloning, expression and characterisation of a membrane progestin receptor and evidence it is an intermediary in meiotic maturation of fish oocytes.Crossref | GoogleScholarGoogle Scholar | 12574519PubMed |
Zhu, H. S., Dou, J. X., Liu, T., and Lin, M. W. (2008). Change of plasma progesterone level of mouse at different phases [In Chinese]. J. Anhui Agri. [Anhui Nongye Kexue] 36, 5901, 5961.