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Positive Therapeutic Effect of Estradiol on Lung Inflammation in Brain Dead Female Rats
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A4734 - Positive Therapeutic Effect of Estradiol on Lung Inflammation in Brain Dead Female Rats
Author Block: F. Yamamoto Ricardo da Silva, R. Armstrong Junior, M. Vidal-dos-Santos, P. Sannomiya, L. F. P. Moreira, A. C. Breithaupt-Faloppa; Laboratório Cirúrgico de Pesquisa Cardiovascular, Instituto do Coracao, Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Sao Paulo, Brazil.
Rationale: Female sex was found to be one of the donor characteristics related with short and long-term survival. We have previously demonstrated that female rats present more severe lung inflammation after brain death (BD) which might be associated with acute estradiol reduction. Here we investigated the potential effects of estradiol treatment in reducing the lung inflammation triggered by BD in female rats. Method: BD was induced, in female Wistar rats, by sudden intracranial balloon inflation and maintained for 6h. Group of rats received 17β-estradiol (E2 50μg/mL i.v. 2mL/h starting after 3 h of BD) and Sham-operated (S) rats were used as controls. Lung vascular permeability was evaluated by Evans blue dye extravasation method. Cell counts, IL-1β and VEGF were quantified in bronchoalveolar lavage (BAL). IL-1β and VEGF concentrations were quantified in lung tissue homogenates and serum. Additionally, lung fragments were maintained for 24 h in culture and IL-1β and VEGF levels were assessed in medium samples. Results: BD increased BAL cell counts, which were reverted after estradiol treatment (S: 17.6±2.5; BD: 36.6±6.4; E2: 17.8±4.7 cells x105/mL; p=0.016), estradiol also reduced IL-1β and increased VEGF in BAL (IL-1β S: 543.2±59.1; BD: 915.1±147.4; E2: 412.0±122.4 pg/mL; p=0.0297; VEGF S: 1263±72.3; BD: 918.1±119.3; E2: 1591±174.1 pg/mL; p=0.006). On lung homogenate IL-1β concentrations increased in BD group (S: 651.5±276; BD: 1979±233.8; E2: 762.4±194 pg/mL/g; p=0.015) and was found elevated in serum and lung culture. Estradiol treated animals also reduced IL-1β levels in serum (S: 58.1±14.1; BD: 124.5±31; E2: 45.5±10.5 pg/mL; p=0.028) and lung culture (S: 1354±508; BD: 9541±4324; E2: 1494±872.4 pg/mL/mg of dry weight; p= 0.0503). Lung vascular permeability was augmented after BD, not being altered by estradiol. Also, VEGF concentrations after BD increased in lung homogenates (S: 228±26; BD: 481±53.4; E2: 470.4±83 pg/mL/mg; p=0.029), however estradiol was not effective in changing its concentrations. Conclusion: Late female donor treatment with estradiol was able to reduce leukocyte infiltration and IL-1β release. Thus this data highlight the estradiol potential for therapeutic application in order to improve donor organ quality. Financial support: grant 2016/03651-0, São Paulo Research Foundation (FAPESP).
Author Block: F. Yamamoto Ricardo da Silva, R. Armstrong Junior, M. Vidal-dos-Santos, P. Sannomiya, L. F. P. Moreira, A. C. Breithaupt-Faloppa; Laboratório Cirúrgico de Pesquisa Cardiovascular, Instituto do Coracao, Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, Sao Paulo, Brazil.
Rationale: Female sex was found to be one of the donor characteristics related with short and long-term survival. We have previously demonstrated that female rats present more severe lung inflammation after brain death (BD) which might be associated with acute estradiol reduction. Here we investigated the potential effects of estradiol treatment in reducing the lung inflammation triggered by BD in female rats. Method: BD was induced, in female Wistar rats, by sudden intracranial balloon inflation and maintained for 6h. Group of rats received 17β-estradiol (E2 50μg/mL i.v. 2mL/h starting after 3 h of BD) and Sham-operated (S) rats were used as controls. Lung vascular permeability was evaluated by Evans blue dye extravasation method. Cell counts, IL-1β and VEGF were quantified in bronchoalveolar lavage (BAL). IL-1β and VEGF concentrations were quantified in lung tissue homogenates and serum. Additionally, lung fragments were maintained for 24 h in culture and IL-1β and VEGF levels were assessed in medium samples. Results: BD increased BAL cell counts, which were reverted after estradiol treatment (S: 17.6±2.5; BD: 36.6±6.4; E2: 17.8±4.7 cells x105/mL; p=0.016), estradiol also reduced IL-1β and increased VEGF in BAL (IL-1β S: 543.2±59.1; BD: 915.1±147.4; E2: 412.0±122.4 pg/mL; p=0.0297; VEGF S: 1263±72.3; BD: 918.1±119.3; E2: 1591±174.1 pg/mL; p=0.006). On lung homogenate IL-1β concentrations increased in BD group (S: 651.5±276; BD: 1979±233.8; E2: 762.4±194 pg/mL/g; p=0.015) and was found elevated in serum and lung culture. Estradiol treated animals also reduced IL-1β levels in serum (S: 58.1±14.1; BD: 124.5±31; E2: 45.5±10.5 pg/mL; p=0.028) and lung culture (S: 1354±508; BD: 9541±4324; E2: 1494±872.4 pg/mL/mg of dry weight; p= 0.0503). Lung vascular permeability was augmented after BD, not being altered by estradiol. Also, VEGF concentrations after BD increased in lung homogenates (S: 228±26; BD: 481±53.4; E2: 470.4±83 pg/mL/mg; p=0.029), however estradiol was not effective in changing its concentrations. Conclusion: Late female donor treatment with estradiol was able to reduce leukocyte infiltration and IL-1β release. Thus this data highlight the estradiol potential for therapeutic application in order to improve donor organ quality. Financial support: grant 2016/03651-0, São Paulo Research Foundation (FAPESP).
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