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. 2020 Oct 14;10:17240. doi: 10.1038/s41598-020-73974-5

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Renal impact at day 1 after hemorrhagic shock (a,b). (a) Tubular injury (KIM-1 mRNA expression, n = 6 for all groups), (b) Blood creatinine (n = 8 for sham and Intact, n = 7 for Ov, n = 6 for Ov + cE2 and n = 7 for Intact + aE2); Renal impact 21 days later (c–f). (c) Glomerular Filtration Rate measured by inulin clearance (n = 9 for sham and Intact, n = 6 for Ov, n = 5 for Ov + cE2 and n = 6 for Intact + aE2), (d) Blood creatinine (n = 6 for all groups), (e) Representative pictures of collagen III staining on kidney sections (magnification × 400, immunohistochemistry, counterstaining with hematoxylin), (f) Quantification of Collagen III expression (immunohistochemistry, n = 5 for each group). Sham: non-shocked mice; Ov: ovariectomized mice; Ov + cE2: ovariectomized mice with chronic estradiol substitution; Intact + V: Non ovariectomized mice receiving vehicle; Intact + aE2: intact mice receiving 25 µg of estradiol at the end of resuscitation. For KIM1 mRNA expression, relative mRNA expression levels were calculated using the ΔΔCtCt method, normalized to HPRT and β actin, and expressed as fold changes relative to the sham animals. Data are presented as mean ± s.e.m. *P < 0.05; **P < 0.01 and ***P < 0.001 versus sham; ^#P < 0.05 and ^##P < 0.01 for indicated comparison.