Metformin prevents endothelial dysfunction through down-regulation of endothelin-1 and upregulation of eNOS expression in the heart of mice with endometriosis.

Please use this identifier to cite or link to this item: https://hdl.handle.net/10216/142262 | Author(s): | Ana Filipa Fernandes Martins | | Title: | Metformin prevents endothelial dysfunction through down-regulation of endothelin-1 and upregulation of eNOS expression in the heart of mice with endometriosis. | | Issue Date: | 2022-05-18 | | Abstract: | Aims: Endometriosis is a gynaecological condition that increases the risk for cardiovascular diseases. This study aims to elucidate whether the anti-diabetic drug metformin modulates morphologic structure, endothelial function, angiogenesis, inflammation, and oxidation-responsive pathways in the heart of mice with surgically-induced endometriosis. Methods and Results: B6CBA/F1 mice (n=37) were divided into Sham(S), Metformin(M), Endometriosis(E) and Metformin/Endometriosis(ME) groups. Cardiomyocytes' cross-sectional area and fibrosis were assessed after Hematoxylin-Eosin and Picrosirius Red staining, respectively. The vasoconstrictor ET-1, nitric oxide synthases-iNOS and eNOS, angiogenesis drivers, VEGF and receptor, VEGFR-2, were detected by immunofluorescence. ET-1, eNOS, VEGF, NF-kB and its inhibitor, Ikβα, involved in inflammatory response, and KEAP-1, an oxidative-stress sensor, were semi-quantified by Western Blotting. MIR199a, MIR16-1, MIR18a, MIR20a, MIR155, MIR200a, MIR342, MIR24-1 and MIR320a that intervene in cardiovascular damage were quantified by Real-Time PCR. Two-way ANOVA test was employed to assess interaction of endometriosis and metformin effects. Cardiomyocyte cross-sectional area increased significantly in M group compared with the other groups. Cardiac fibrosis increased with endometriosis; E and ME groups showed increased fibrosis relatively to controls. The association of endometriosis and metformin significantly influenced ET-1 and eNOS expression; ME group presented lower ET-1 and higher eNOS expression than E group. ME group showed a decreasing tendency of VEGF levels compared to E group. Endometriosis significantly incremented NF-kB expression, which metformin failed to mitigate. Neither Ikβα nor KEAP-1 varied with disease or treatment. The expression of MIR199a, MIR16-1, and MIR18a significantly decreased with endometriosis whereas MIR20a showed an equivalent trend, globally reducing cardioprotection. MIR155, 342, 20a and 24-1 did not vary among groups but tended to decrease in E and ME groups. In contrast, MIR320a showed a tendency to decrease in metformin-treated groups. Conclusion: Metformin diminished endometriosis-associated endothelial dysfunction but failed to mitigate the increase in NF-kB expression and cardiac fibrosis observed in mice with endometriosis. | | Description: | Objetivos: A endometriose associa-se ao aumento do risco de doença cardiovascular. O objetivo deste estudo é avaliar se o fármaco antidiabético metformina é capaz de modular a estrutura morfológica, a função endotelial, a angiogénese e as vias de inflamação e as associadas à resposta à oxidação no coração do murganho com endometriose induzida cirurgicamente. Métodos e Resultados: Murganhos B6CBA/F1 (n=37) foram divididos nos grupos Sham(S), Metformina(M), Endometriose(E) e Metformina/Endometriose(ME). A área dos cardiomiócitos e a fibrose cardíaca foram avaliadas após coloração com Hematoxilina/Eosina e Picrosirius Red, respetivamente. O vasoconstritor, ET-1, as síntases de óxido nítrico, iNOS e eNOS, o VEGF e o seu recetor, VEGFR-2, efetores na angiogénese, foram detetados por imunofluorescência. A ET-1, eNOS, VEGF, NF-kB e o seu inibidor, Ikβα, intervenientes na resposta inflamatória, e o sensor de stress-oxidativo KEAP-1, foram semi-quantificados por Western Blotting. Os microRNAs MIR199a, MIR16-1, MIR18a, MIR20a, MIR155, MIR200a, MIR342, MIR24-1 e MIR320a, intervenientes no dano cardiovascular, foram quantificados por PCR em Tempo Real. A análise two-way ANOVA foi efetuada para avaliar a interação entre a endometriose e o tratamento com metformina. A área dos cardiomiócitos aumentou significativamente nos animais do grupo M relativamente aos outros grupos. A fibrose cardíaca aumentou com a endometriose; nos grupos E e ME houve aumento quando comparados com os controlos. A associação da endometriose e do tratamento com metformina influenciou significativamente a expressão da ET-1 e da eNOS; a ET-1 diminui e a eNOS no grupo ME relativamente a E. O VEGF mostrou tendência para diminuir em ME comparativamente a E. A endometriose aumentou significativamente a expressão de NF-kB que não reverteu com o tratamento com metformina. A expressão de Ikβα e KEAP-1 não variou com a doença ou com tratamento. A expressão dos MIR199a, MIR16-1e MIR18a diminuiu significativamente na endometriose; o MIR20a mostrou tendência equivalente, o que globalmente reduziu a cardioproteção. Os MIR155, 342, 20a e 24-1 não variaram, mas tenderam a diminuir nos grupos E e ME. O MIR320a, pelo contrário, mostrou tendência a diminuir nos grupos tratados com metformina. Conclusão: A metformina diminuiu a disfunção endotelial associada à endometriose, mas não conseguiu mitigar o aumento da expressão de NF-kB, ou, reverter a fibrose cardíaca observada em murganhos com endometriose. | | Subject: | Ciências da saúde Health sciences | | Scientific areas: | Ciências médicas e da saúde::Ciências da saúde Medical and Health sciences::Health sciences | | DOI: | 10.34626/hwsr-zt21 | | TID identifier: | 203176294 | | URI: | https://hdl.handle.net/10216/142262 | | Document Type: | Dissertação | | Rights: | restrictedAccess | | License: | https://creativecommons.org/licenses/by-nc-nd/4.0/ | | Appears in Collections: | FMUP - Dissertação | Files in This Item: | File | Description | Size | Format | | |---|---|---|---|---| | 569441.pdf Restricted Access | Metformin prevents endothelial dysfunction through down-regulation of endothelin-1 and upregulation of eNOS expression in the heart of mice with endometriosis. | 10.23 MB | Adobe PDF | View/Open | This item is licensed under a Creative Commons License