Role of ENaC in gender-associated differences in blood pressure

Document Type : Original Article

Authors

1 Department of Clinical Laboratory, Children’s Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China, 310052

2 Qiaosi Branch, First People’s Hospital of Linping District, Hangzhou, Zhejiang Province, China

Abstract

Objective(s): Sexual dimorphism in blood pressure regulation has been extensively noted in humans, but the underlying mechanisms remain to be fully understood. Our research aims to investigate the possible correlation between gender-associated differences in blood pressure and renal sodium transport. 
Materials and Methods: We measured male and female mice’s blood pressure, urine, and plasma sodium concentration when fed a regular or high-Na+ diet. After that, their renal sodium transporters were assessed by western blot and immunofluorescence. For further investigation, male mice were castrated to observe the differences in blood pressure and renal sodium transporters compared to normal mice. 
Results: Male mice exhibited higher blood pressure and lower renal sodium excretion than female littermates. Furthermore, the blood pressure of male mice exhibited a more significant and rapid increase relative to female mice when the diet was switched from control sodium to high sodium. Western blot and immunofluorescent staining revealed that in male mice, the sodium transporters epithelial sodium channel (ENaC) and the upstream kinases SPAK (Ste20-related proline/alanine-rich kinase), OSR1 (oxidative stress response kinase 1), and WNK4 (Lysine-Deficient Protein Kinase 4) were elevated. Beyond that, male mice exhibited lowered blood pressure and reduced abundance of ENaC (α, β, and γ) after castration.
Conclusion: ENaC plays a significant role in gender-associated differences in blood pressure and renal sodium reabsorption.

Keywords

Main Subjects

References

1. Samora M, Incognito AV, Vianna LC. Sex differences in blood pressure regulation during ischemic isometric exercise: The role of the β-adrenergic receptors. J Appl Physiol 2019; 127:408-414.
2. Padilha MCSV, Ferreira FC, Oliveira TLS, Soares PPS, Rodrigues GD. Sex differences in blood pressure regulation during the isometric exercise under heated environment. Blood Press Monit 2022; 27:55-62.
3. Gerdts E, Sudano I, Brouwers S, et al. Sex differences in arterial hypertension. Eur Heart J 2022; 43:4777-4788.
4. Elmarakby AA, Sullivan JC. Sex differences in hypertension: Lessons from spontaneously hypertensive rats (SHR). Clin Sci 2021; 135:1791-1804.
5. Gillis EE, Belanger K, Abdelbary M, Mohamed R, Sun J, Brands MW, et al. Splenectomy increases blood pressure and abolishes sex differences in renal T-regulatory cells in spontaneously hypertensive rats. Clin Sci 2021; 135:2329-2339.
6. Azurmendi PJ, Toro AR, Celía AF, Guevara D, Solerno MR, Di Ciano LA, et al. Behavior of the renal kallikrein in spontaneously hypertensive rats: Influence of sexual hormones and aldosterone-sensitive distal nephron ion channels. Peptides 2023; 160:170925.
7. Yu G, Cheng M, Wang W, Zhao R, Liu Z. Involvement of WNK1-mediated potassium channels in the sexual dimorphism of blood pressure. Biochem Biophys Res Commun 2017; 485:255-260.
8. Blanchard A, Bockenhauer D, Bolignano D, Calo LA, Cosyns E, Devuyst O, et al. Gitelman syndrome: Consensus and guidance from a kidney disease: Improving Global Outcomes (KDIGO) Controversies Conference. Kidney Int 2017; 91:24–33.
9. Viering D, Schlingmann KP, Hureaux M, Nijenhuis T, Mallett A, Chan MMY, et al. Gitelman-like syndrome caused by pathogenic variants in mtDNA. J Am Soc Nephrol 2022; 33:305-325.
10. Liu Z, Xie J, Wu T, Truong T, Auchus RJ, Huang CL. Down-regulation of NCC and NKCC2 cotransporters by kidney-specific WNK1 revealed by gene disruption and transgenic mouse models. Hum Mol Genet 2011; 20:855-866.
11. Li Z, Wu H, Wei S, Liu M, Shi Y, Li M, et al. R158Q and G212S, novel pathogenic compound heterozygous variants in SLC12A3 of Gitelman syndrome. Front Med 2022; 16:932-945.
12. Chiga M, Rafiqi FH, Alessi DR, Sohara1 E, Ohta1A, Rai1T, et al. Phenotypes of pseudohypoaldosteronism type II caused by the WNK4 D561A missense mutation are dependent on the WNK-OSR1/SPAK kinase cascade. J Cell Sci 2011; 124:1391-1395.
13. Puleo F, Kim K, Frame AA, Walsh KR, Ferdaus MZ, Moreira JD, et al. Sympathetic regulation of the NCC (sodium chloride cotransporter) in dahl salt-sensitive hypertension. Hypertension 2020; 76:1461-1469.
14. Gallafassi EA, Bezerra MB, Reboucas NA. Control of sodium and potassium homeostasis by renal distal convoluted tubules. Braz J Med Biol Res 2023; 56: e12392.
15. Hager H, Kwon TH, Vinnikova AK, Masilamani S, Brooks HL, Frokiaer J, et al. Immunocytochemical and immunoelectron microscopic localization of α-, β-, and γ-ENaC in rat kidney. Am J Physiol Ren Physiol 2001; 280: F1093-F1106.
16 Rotin D, Staub O. Function and regulation of the epithelial Na+ channel ENaC. Compr Physiol 2021; 11:2017-2045. 
17. Rotin D, Schild L. ENaC and its regulatory proteins as drug targets for blood pressure control. Curr Drug Targets 2008; 9:709-716.
18. McDonald FJ, Yang B, Hrstka RF, Drummond HA, Tarr DE, McCray PB, et al. Disruption of the β subunit of the epithelial Na+ channel in mice: Hyperkalemia and neonatal death associated with a pseudohypoaldosteronism phenotype. Proc Natl Acad Sci USA 1999; 96:1727-1731.
19. Pham TD, Elengickal AJ, Verlander JW, Al-Qusairi L, Chen C, Abood DC, et al. Pendrin-null mice develop severe hypokalemia following dietary Na+ and K+ restriction: role of ENaC. Am J Physiol Renal Physiol 2022; 322:F486-F497.
20. Quinkler M, Bujalska IJ, Kaur K, Onyimba CU, Buhner S, Allolio B, et al. Androgen receptor–mediated regulation of the α-subunit of the epithelial sodium channel in human kidney. Hypertension 2005; 46:787-798.
21. Heo N, Son MJ, Lee J, Jung J, Kim S, Oh Y, et al. Effect of estradiol on the expression of renal sodium transporters in rats. Climacteric 2013; 16:265-273.
22. Maeoka Y, Nguyen LT, Sharma A, Cornelius RJ, Su XT, Gutierrez MR, et al. Dysregulation of the WNK4-SPAK/OSR1 pathway has a minor effect on baseline NKCC2 phosphorylation. Am J Physiol Renal Physiol 2024; 326:F39-F56.
23. Lin SC, Ma C, Chang KJ, Cheong HP, Lee MC, Lan YT, et al. The post-translational modification networking in WNK-centric hypertension regulation and electrolyte homeostasis. Biomedicines 2022; 10:2169-2205.
24. Ahmed M, Salker MS, Elvira B, Umbach AT, Fakhri H, Saeed AM, et al. SPAK sensitive regulation of the epithelial Na+ channel ENaC. Kidney Blood Press Res 2015; 40:335-343.
25. Uchida S, Mori T, Susa K, Sohara E. NCC regulation by WNK signal cascade. Front Physiol 2023; 13:1081261.
26. Nakano K, Kubota Y, Mori T, Chiga M, Mori T, Sonoda S, et al. Familial cases of pseudohypoaldosteronism type II harboring a novel mutation in the Cullin 3 gene. Nephrology 2020; 25:818-821. 
27. Brown A, Meor Azlan NF, Wu Z, Zhang J. WNK-SPAK/OSR1-NCC kinase signaling pathway as a novel target for the treatment of salt-sensitive hypertension. Acta Pharmacol Sin 2021; 42:508-517.
28. Rafiqi FH, Zuber AM, Glover M, Richardson C, Fleming S, Jovanović S, et al. Role of the WNK‐activated SPAK kinase in regulating blood pressure. EMBO Mol Med 2010; 2:63-75.
29. Harrison DG, Bader M, Lerman LO, Fink G, Karumanchi SA, Reckelhoff JF, et al. Tail-cuff versus radiotelemetry to measure blood pressure in mice and rats. Hypertension 2024; 81:3-5.
30. Mutchler SM, Kleyman TR. Effects of amiloride on acetylcholine-dependent arterial vasodilation evolve over time in mice on a high salt diet. Physiol Rep 2022; 10:e15255.
31. Yu Q, Li J, Dai CL, Li H, Iqbal K, Liu F, et al. Anesthesia with sevoflurane or isoflurane induces severe hypoglycemia in neonatal mice. PLoS One 2020; 15:e0231090.
32. Wilhelmson AS, Johansson I, Fogelstrand L, Fagman JB, Arnal JF, Karlsson MCI, et al. Deficiency of mature B cells does not alter the atherogenic response to castration in male mice. Sci Rep 2022; 12:12931-12936.
33. Hadchouel J, Soukaseum C, Büsst C, Zhou XO, Baudrie V, Zürrer T, et al. Decreased ENaC expression compensates the increased NCC activity following inactivation of the kidney-specific isoform of WNK1 and prevents hypertension. Proc Natl Acad Sci USA 2010; 107:18109-18114.
34. Vidal-Petiot E, Elvira-Matelot E, Mutig K, Soukaseum C, Baudrie V, Wu S, et al. WNK1-related familial hyperkalemic hypertension results from an increased expression of L-WNK1 specifically in the distal nephron. Proc Natl Acad Sci USA 2013; 110:14366-14371.
35. Chávez-Canales M, Zhang C, Soukaseum C, Moreno E, Pacheco-Alvarez D, Vidal-Petiot E, et al. WNK-SPAK-NCC cascade revisited WNK1 stimulates the activity of the Na-Cl cotransporter via SPAK, an effect antagonized by WNK4. Hypertension 2014; 64:1047-1053.
36. Carmosino M, Rizzo F, Ferrari P, Torielli L, Ferrandi M, Bianchi G, et al. NKCC2 is activated in Milan hypertensive rats contributing to the maintenance of salt-sensitive hypertension. Pflugers Arch 2011; 462:281-291.
37. Mutchler SM, Kirabo A, Kleyman TR. Epithelial sodium channel and salt-sensitive hypertension. Hypertension 2021; 77:759-767.
38. Yang SS, Lo YF, Wu CC, Lin SW, Yeh CJ, Chu P, et al. SPAK-knockout mice manifest Gitelman syndrome and impaired vasoconstriction. J Am Soc Nephrol 2010; 21:1868-1877.
39. Susa K, Sohara E, Rai T, Zeniya M, Mori Y, Mori T, et al. Impaired degradation of WNK1 and WNK4 kinases causes PHAII in mutant KLHL3 knock-in mice. Hum Mol Genet 2014; 23:5052-5060.
40. Takahashi D, Mori T, Nomura N, Khan MZH, Araki Y, Zeniya M, et al. WNK4 is the major WNK positively regulating NCC in the mouse kidney. Biosci Rep 2014; 34: e00107.
41. Vitari AC, Deak M, Morrice NA, Alessi DR. The WNK1 and WNK4 protein kinases that are mutated in Gordon’s hypertension syndrome phosphorylate and activate SPAK and OSR1 protein kinases. Biochem J 2005; 391:17-24.
42. Hoorn EJ, de Baaij JHF. Chloride-sensitive signaling turns the potassium switch on. Kidney Int 2022; 102:956-958. 
43. Mayan H, Melnikov S, Novikov I, Holtzman EJ, Farfel Z. Familial hyperkalemia and hypertension: pathogenetic insights based on lithium clearance. J Clin Endocr Metab 2009; 94:3010-3016.
44. Zhang Y, Li C, Li W, Zhao Y. Estrogen regulation of human with-no-lysine (K) kinase-4 gene expression involves AP-1 transcription factor. Mol Cell Endocrinol 2011; 332:140-148.
Iranian Journal of Basic Medical Sciences
Volume 28, Issue 4
April 2025
Pages 527-532

Files

Share

How to cite

Statistics