Açıklanamayan infertilite ve endometriozisli infertil hastaların mid-sekretuar endometriyumunda TRPV6 ve PMCA1’in ekspresyonu

Year 2023, Volume: 48 Issue: 3, 815 - 824, 30.09.2023

Tuba Ozcan Metin , Nafiye Yılmaz , Banu Yılmaz , Şakir Necat Yılmaz , Mehmet Emin Erdal , İlay Buran , Gülhan Temel , Adem Doğaner

https://doi.org/10.17826/cumj.1310778

Abstract

Amaç: Bu çalışmanın amacı, açıklanamayan infertilite (Aİ) ve endometriozisi (E) olan infertil kadınların endometriyumunda Transient Reseptör Potansiyel Vanilloid 6 (TRPV6) ve Plazma Membran Ca2+ ATPaz 1 (PMCA1) ekspresyonunu değerlendirmek ve bulguları sağlıklı fertil kadınlarla karşılaştırmaktır.
Gereç ve Yöntem: TRPV6 ve PMCA1 proteinlerinin endometriyal ekspresyonu mid-sekretuar faz sırasında, immünohistokimyasal yöntemle değerlendirilmiş ve mRNA seviyeleri real-time PCR ile ölçülmüştür. Kalsiyum konsantrasyonları İndüktif Eşleşmiş Plazma-Kütle Spektrometrisi (ICP-MS) kullanılarak belirlenmiştir.
Bulgular: TRPV6 ve PMCA1 protein ekspresyonu kontrol grubu ile karşılaştırıldığında, Aİ ve E gruplarının hem luminal hem de glandular epitelinde anlamlı olarak artış saptanmıştır. TRPV6 mRNA ekspresyon seviyeleri Aİ grubunda (0.525 ± 0.087) kontrol grubuna (0.809 ± 0.073) kıyasla anlamlı derecede düşüktür. PMCA1 mRNA ekspresyon düzeylerinde gruplar arasında anlamlı bir fark yoktur. Kalsiyum konsantrasyonu Aİ’li kadınlarda (medyan (Q1-Q3): 50.57 [40.67-51.37], kontrol grubuna (medyan (Q1-Q3): 24.29 [20.11-25.55]) göre anlamlı derecede yüksek bulunmuştur.
Sonuç: Mevcut bulgular, implantasyon penceresi sırasında özellikle Aİ’li hastalarda, kalsiyum konsantrasyonlarındaki farklılıkların yanı sıra, TRPV6 ve PMCA1 mRNA ve proteinlerinin değişen ekspresyonlarını göstermektedir. Bu farklılıklar, potansiyel olarak infertilitenin altında yatan nedenlerden biri olarak düşünülebilir. Bu kanalların fonksiyonel rollerini aydınlatmak ve infertilite tedavisinde farmasötik yaklaşımda yeni stratejiler geliştirmek için daha fazla çalışmaya ihtiyaç vardır.

Keywords

Kalsiyum metabolizması, endometriozis, TRPV6, PMCA1, açıklanamayan infertilite

The expression of TRPV6 and PMCA1 in the mid-secretory endometrium of infertile patients with unexplained infertility and endometriosis

Abstract

Purpose: The goal of this study was to evaluate the expression of transient receptor potential vanilloid 6 (TRPV6) and plasma membrane Ca2+ ATPase 1 (PMCA1) in the endometrium of infertile women with unexplained infertility (UI) and endometriosis (E), and to compare the findings to healthy fertile women.
Materials and Methods: Endometrial expression of TRPV6 and PMCA1 proteins was evaluated during the mid-secretory phase using the immunohistochemical method, and mRNA levels were measured by real-time PCR. Calcium concentrations were determined using Inductively coupled plasma-mass spectrometry (ICP-MS).
Results: TRPV6 and PMCA1 protein expression were significantly increased in both luminal and glandular epithelium of UI or E groups compared to the control group. TRPV6 mRNA expression levels were significantly lower in the UI group (0.525 ± 0.087) compared to the control group (0.809 ± 0.073). There was no significant difference in PMCA1 mRNA expression levels between the groups. Calcium concentration was significantly higher in women with UI (median (Q1-Q3): 50.57 [40.67-51.37], compared to the control group (median (Q1-Q3): 24.29 [20.11-25.55]).
Conclusion: The present findings demonstrate altered expressions of TRPV6 and PMCA1 mRNA and protein, along with differences in calcium concentration among patients, especially those with the UI during the window of implantation. These differences might potentially be considered as one of the underlying causes of infertility. Further studies are needed to elucidate the functional roles of these channels and to develop new strategies for pharmaceutical intervention in the treatment of infertility.

Keywords

Calcium metabolism, endometriosis, TRPV6, PMCA1, unexplained infertility

References

• Singh M, Chaudhry P, Asselin E. Bridging endometrial receptivity and implantation: network of hormones, cytokines, and growth factors. J Endocrinol. 2011;210:5-14.
• Ruan YC, Chen H, and Chan HC. Ion channels in the endometrium: regulation of endometrial receptivity and embryo implantation. Hum Reprod Update. 2014;20:517-29.
• Clapham DE. Calcium signaling. Cell. 2007;131:1047-58.
• Baczyk D, Kingdom JC, Uhlén P. Calcium signaling in placenta. Cell Calcium. 2011;49:350-56.
• Uhlén P, Fritz N. Biochemistry of calcium oscillations. Biochem Biophys Res Commun. 2010;96:28-32.
• Nilius B, Owsianik G, Voets T, Peters JA. Transient receptor potential cation channels in disease. Physiol Rev. 2007;87:165-17.
• Choi Y, Seo H, Kim M, Ka H. Dynamic expression of calcium-regulatory molecules, TRPV6 and S100G, in the uterine endometrium during pregnancy in pigs. Biol Reprod. 2009;81:1122-30.
• Bianco SD, Peng JB, Takanaga H, Suzuki Y, Crescenzi A, Kos CH et al. Marked disturbance of calcium homeostasis in mice with targeted disruption of the Trpv6 calcium channel gene. J Bone Miner Res. 2007;22:274-85.
• Brini M, Calì T, Ottolini D, Carafoli E. The plasma membrane calcium pump in health and disease. FEBS J. 2013;280:5385-97.
• Kim HJ, Lee GS, Ji YK, Choi KC, Jeung EB. Differential expression of uterine calcium transporter 1 and plasma membrane Ca2+ ATPase 1b during rat estrous cycle. Am J Physiol Endocrinol Metab. 2006;291:E234-41.
• Choi Y, Seo H, Shim J, Yoo I, Ka H. Calcium extrusion regulatory molecules: differential expression during pregnancy in the porcine uterus. Domest Anim Endocrinol. 2014;47:1-10.
• Yang H, Choi KC, Hyun SH, Jeung EB. Coexpression and estrogen-mediated regulation of TRPV6 and PMCA1 in the human endometrium during the menstrual cycle. Mol Reprod Dev. 2011;78:274-82.
• Kobayashi Y, Hirawa N, Tabara Y, Muraoka H, Fujita M, Miyazaki N et al. Mice lacking hypertension candidate gene ATP2B1 in vascular smooth muscle cells show significant blood pressure elevation. Hypertension. 2012;59:854-60.
• Werbrouck E, Spiessens C, Meuleman C, D’Hooghe T. No difference in cycle pregnancy rate and in cumulative live-birth rate between women with surgically treated minimal to mild endometriosis and women with unexplained infertility after controlled ovarian hyperstimulation and intrauterine insemination. Fertil Steril. 2006;86:566-71.
• Noyes RW, Hertig AT, Rock J. Dating the endometrial biopsy. Am J Obstet Gynecol. 1975; 122:262-63.
• Beavis A, Dawson M, Doble P, Scolyer RA, Bourne R, Li LX. Confirmation of sentinel lymph node identity by analysis of fine-needle biopsy samples using inductively coupled plasma-mass spectrometry. Ann Surg Oncol. 2008;15:934-40.
• Jha RK, Titus S, Saxena D, Kumar PG, Laloraya M. Profiling of E-cadherin, beta-catenin and Ca(2+) in embryo-uterine interactions at implantation. FEBS Lett. 2006;580:5653-60.
• Wang J, Mayernik L, Armant DR. Integrin signaling regulates blastocyst adhesion to fibronectin at implantation: intracellular calcium transients and vesicle trafficking in primary trophoblast cells. Dev Biol. 2002;245:270-79.
• Singh H, Aplin JD. Adhesion molecules in endometrial epithelium: tissue integrity and embryo implantation. J Anat. 2009;215:3-13.
• Choi Y, Seo H, Kim M, Ka H. Dynamic expression of calcium-regulatory molecules, TRPV6 and S100G, in the uterine endometrium during pregnancy in pigs. Biol Reprod. 2009;81:1122-30.
• Bhusane K, Bhutada S, Chaudhari U, Savardekar L, Katkam R, Sachdeva G. Secrets of endometrial receptivity: Some are hidden in uterine secretome. Am J Reprod Immunol. 2016; 75:226-36.
• Pang ZP, Südhof TC. Cell biology of Ca2+-triggered exocytosis. Curr Opin Cell Biol. 2010;22:496-505.
• Spencer TE. Biological roles of uterine glands in pregnancy. Semin Reprod Med. 2014;32:346-57.
• Yang H, Kim TH, An BS, Choi KC, Lee HH, Kim JM et al. Differential expression of calcium transport channels in placenta primary cells and tissues derived from preeclamptic placenta. Mol Cell Endocrinol. 2013;367:21-30.
• Yang H, An BS, Choi KC, Jeung EB. Change of genes in calcium transport channels caused by hypoxic stress in the placenta, duodenum, and kidney of pregnant rats. Biol Reprod. 2013;88:30.
• Toledo Mauriño-JJ, Fonseca-Camarillo G. TRPV subfamily (TRPV2, TRPV3, TRPV4, TRPV5, and TRPV6) gene and protein expression in patients with ulcerative colitis. J Immunol Res. 2020;2020:2906845.
• Vachel L, Norez C, Jayle C, Becq F, Vandebrouck C. The low PLC-δ1 expression in cystic fibrosis bronchial epithelial cells induces upregulation of TRPV6 channel activity. Cell Calcium. 2015;57:38-48.
• Carafoli E, Santella L, Branca D, Brini M. Generation, control, and processing of cellular calcium signals. Crit Rev Biochem Mol Biol. 2001;36:107-260.
• Choi KC, An BS, Yang H, Jeung EB. Regulation and molecular mechanisms of calcium transport genes: do they play a role in calcium transport in the uterine endometrium? J Physiol Pharmacol. 2011;62:499-504.
• Giacomello M, De Mario A, Scarlatti C. Plasma membrane calcium ATPases and related disorders. Int J Biochem Cell Biol. 2013;45:753-62.
• Jiang L, Allagnat F, Nguidjoe E, Kamagate A, Pachera N, Vanderwinden JM et al. Plasma membrane Ca2+-ATPase overexpression depletes both mitochondrial and endoplasmic reticulum Ca2+ stores and triggers apoptosis in insulin-secreting BRIN-BD11 cells. J Biol Chem. 2010;285:30634-43.
• Marian MJ, Mukhopadhyay P, Borchman D, Tang D, Paterson CA. The effect of hydrogen peroxide on sarco/endoplasmic and plasma membrane calcium ATPase gene expression in cultured human lens epithelial cells. Open Ophthalmol J. 2008;2:123-29.
• Maléth J, Hegyi P. Ca2+ toxicity and mitochondrial damage in acute pancreatitis: translational overview. Philos Trans R Soc Lond B Biol Sci. 2016;371:20150425.
• De Clercq K, Held K, Van Bree R, Meuleman C, Peeraer K, Tomassetti C et al. Functional expression of transient receptor potential channels in human endometrial stromal cells during the luteal phase of the menstrual cycle. Hum Reprod. 2015;30:1421-36.
• De Clercq K, Van den Eynde C, Hennes A, Van Bree R, Voets T, Vriens J. The functional expression of transient receptor potential channels in the mouse endometrium. Hum Reprod. 2017;32:615-30.
• Stark AM, Pfannenschmidt S, Tscheslog H, Maass N, Rösel F, Mehdorn HM et al. Reduced mRNA and protein expression of BCL-2 versus decreased mRNA and increased protein expression of BAX in breast cancer brain metastases: a real-time PCR and immunohistochemical evaluation. Neurol Res. 2006;28:787-93.
• Sarro SM, Unruh TL, Zuccolo J, Sanyal R, Luider JM, Auer-Grzesiak IA et al. Quantification of CD20 mRNA and protein levels in chronic lymphocytic leukemia suggests a post-transcriptional defect. Leuk Res. 2010;34:1670-3.
• Lichtinghagen R, Musholt PB, Lein M, Römer A, Rudolph B, Kristiansen G et al. Different mRNA and protein expression of matrix metalloproteinases 2 and 9 and tissue inhibitor of metalloproteinases 1 in benign and malignant prostate tissue. Eur Urol. 2002;42:398-406.
• Zhao BS, Roundtree IA, He C. Post-transcriptional gene regulation by mRNA modifications. Nat Rev Mol Cell Biol. 2017;18:31-42.
• Guyot E, Solovyova Y, Tomkiewicz C, Leblanc A, Pierre S, El Balkhi S et al. Determination of Heavy Metal Concentrations in Normal and Pathological Human Endometrial Biopsies and In Vitro Regulation of Gene Expression by Metals in the Ishikawa and Hec-1b Endometrial Cell Line. PLoS One. 2015;10:e0142590.
• Yaman M, Kaya G, Simsek M. Comparison of trace element concentrations in cancerous and noncancerous human endometrial and ovary tissues. Int J Gynecol Cancer. 2007;17:220-8.
• Nasiadek M, Krawczyk T, Sapota A. Tissue levels of cadmium and trace elements in patients with myoma and uterine cancer. Hum Exp Toxicol. 2005;24:623-30