The SGLT-2 inhibitor empagliflozin improves myocardial strain, reduces cardiac fibrosis and pro-inflammatory cytokines in non-diabetic mice treated with doxorubicin.
The SGLT-2 inhibitor empagliflozin improves myocardial strain, reduces cardiac fibrosis and pro-inflammatory cytokines in non-diabetic mice treated with doxorubicin.
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DOI：
10.1186/s12933-021-01346-y
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发表时间：
2021-07-23
影响因子：
9.3
通讯作者：
Maurea N
中科院分区：
文献类型：
--
作者：
Quagliariello V;De Laurentiis M;Rea D;Barbieri A;Monti MG;Carbone A;Paccone A;Altucci L;Conte M;Canale ML;Botti G;Maurea N
Empagliflozin (EMPA) 是一种钠葡萄糖协同转运蛋白 2 的选择性抑制剂，在 EMPA-REG OUTCOME 试验中可降低 2 型糖尿病患者因心力衰竭和心血管死亡住院的风险。最近的试验证明，EMPA 通过参与仍有待深入分析的生化途径，对非糖尿病患者具有多种心肾益处。我们的目的是通过分析 NLRP3 炎性体和 MyD88 相关通路，评估 EMPA 对阿霉素 (DOXO) 治疗的非糖尿病小鼠心肌应变的影响，从而产生抗凋亡和抗纤维化作用。对单独暴露于阿霉素或联合 EMPA 的小鼠心肌细胞（HL-1 细胞系）进行了初步细胞研究。进行了以下分析：细胞活力测定（通过改良的 MTT 测定）、细胞内 ROS 产生的研究、脂质过氧化（定量细胞内丙二醛和 4-羟基壬烯醛）、细胞内 Ca2+ 稳态。此外，还进行了促炎症研究：NLRP3 炎性小体、MyD88 myddosome 和 p65/NF-κB 的表达与心脏毒性相关细胞因子（白细胞介素 1β、8、6）的分泌相关。 C57Bl/6 小鼠未经治疗 (Sham, n = 6) 或用阿霉素 (DOXO, n = 6)、EMPA (EMPA, n = 6) 或阿霉素联合 EMPA (DOXO-EMPA, n = 6) 治疗 10 天。 DOXO 腹腔注射。在治疗前后研究了铁死亡和黄嘌呤氧化酶。通过经胸超声心动图 (Vevo 2100) 分析心脏功能研究，包括 EF、FS 和径向/纵向应变。分别通过天狼星红和 TUNEL 测定对心脏纤维化和细胞凋亡进行组织学研究，并通过前胶原蛋白 1α1、MMP-9 和 Caspase-3 表达进行定量。还通过 ELISA 方法对治疗前后的组织 NLRP3、MyD88 和细胞因子进行了定量。暴露于阿霉素的心肌细胞增加了细胞内 Ca2+ 含量以及与细胞死亡相关的几种促炎标记物的表达；与 EMPA 共孵育显着降低了影响的程度。在临床前研究中，与 DOXO 组相比，EMPA 增加了 EF 和 FS (p<0.05)，并防止了用阿霉素 (RS) 治疗 10 天后径向和纵向应变的减少，EMPA-DOXO 小鼠为 30.3%，而 DOXO 小鼠为 15.7%； EMPA-DOXO 中的 LS − 17%，而 DOXO 小鼠中的 LS – 11.7%（两者的 p < 0.001）。与 DOXO 相关的 EMPA 中铁死亡、黄嘌呤氧化酶表达、心脏纤维化和细胞凋亡也显着减少。与 DOXO 相比，DOXO-EMPA 组的心脏、肝脏和肾脏中促炎细胞因子、NLRP3、MyD88 和 NF-kB 的表达也有所减少 (p<0.001)。 EMPA 通过参与 NLRP3 和 MyD88 相关通路，减少阿霉素治疗小鼠的铁死亡、纤维化、细胞凋亡和炎症，从而显着改善心脏功能。这些发现为旨在减少接受阿霉素治疗的非糖尿病癌症患者的不良心血管结局的转化研究提供了概念证明。
Empagliflozin (EMPA), a selective inhibitor of the sodium glucose co-transporter 2, reduced the risk of hospitalization for heart failure and cardiovascular death in type 2 diabetic patients in the EMPA‐REG OUTCOME trial. Recent trials evidenced several cardio-renal benefits of EMPA in non-diabetic patients through the involvement of biochemical pathways that are still to be deeply analysed. We aimed to evaluate the effects of EMPA on myocardial strain of non-diabetic mice treated with doxorubicin (DOXO) through the analysis of NLRP3 inflammasome and MyD88-related pathways resulting in anti-apoptotic and anti-fibrotic effects. Preliminary cellular studies were performed on mouse cardiomyocytes (HL-1 cell line) exposed to doxorubicin alone or combined to EMPA. The following analysis were performed: determination of cell viability (through a modified MTT assay), study of intracellular ROS production, lipid peroxidation (quantifying intracellular malondialdehyde and 4-hydroxynonenal), intracellular Ca2+ homeostasis. Moreover, pro-inflammatory studies were also performed: expression of NLRP3 inflammasome, MyD88 myddosome and p65/NF-κB associated to secretion of cytokines involved in cardiotoxicity (Interleukins 1β, 8, 6). C57Bl/6 mice were untreated (Sham, n = 6) or treated for 10 days with doxorubicin (DOXO, n = 6), EMPA (EMPA, n = 6) or doxorubicin combined to EMPA (DOXO-EMPA, n = 6). DOXO was injected intraperitoneally. Ferroptosis and xanthine oxidase were studied before and after treatments. Cardiac function studies, including EF, FS and radial/longitudinal strain were analysed through transthoracic echocardiography (Vevo 2100). Cardiac fibrosis and apoptosis were histologically studied through Picrosirius red and TUNEL assay, respectively and quantified through pro-collagen-1α1, MMP-9 and Caspase-3 expression. Tissue NLRP3, MyD88 and cytokines were also quantified before and after treatments through ELISA methods. Cardiomyocytes exposed to doxorubicin increased the intracellular Ca2+ content and expression of several pro-inflammatory markers associated to cell death; co-incubation with EMPA reduced significantly the magnitude of the effects. In preclinical study, EMPA increased EF and FS compared to DOXO groups (p < 0.05), prevented the reduction of radial and longitudinal strain after 10 days of treatment with doxorubicin (RS) 30.3% in EMPA-DOXO vs 15.7% in DOXO mice; LS − 17% in EMPA-DOXO vs – 11.7% in DOXO mice (p < 0.001 for both). Significant reductions in ferroptosis, xanthine oxidase expression, cardiac fibrosis and apoptosis in EMPA associated to DOXO were also seen. A reduced expression of pro-inflammatory cytokines, NLRP3, MyD88 and NF-kB in heart, liver and kidneys was also seen in DOXO-EMPA group compared to DOXO (p < 0.001). EMPA reduced ferroptosis, fibrosis, apoptosis and inflammation in doxorubicin-treated mice through the involvement of NLRP3 and MyD88-related pathways, resulting in significant improvements in cardiac functions. These findings provides the proof of concept for translational studies designed to reduce adverse cardiovascular outcomes in non-diabetic cancer patients treated with doxorubicin.
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影响因子：
3.2
作者：
Mascarenhas, Leo;Malogolowkin, Marcio;Armenian, Saro H.;Sposto, Richard;Venkatramani, Rajkumar
通讯作者：
Venkatramani, Rajkumar
DOI：
10.1093/gerona/gly139
发表时间：
2019-03-14
期刊：
The journals of gerontology. Series A, Biological sciences and medical sciences
影响因子：
--
作者：
de Lucia C;Wallner M;Eaton DM;Zhao H;Houser SR;Koch WJ
通讯作者：
Koch WJ
影响因子：
3.9
作者：
Di Stadio, Chiara Stella;Altieri, Filomena;Arcari, Paolo
通讯作者：
Arcari, Paolo
影响因子：
11.2
作者：
Lyu, Yi Lisa;Kerrigan, John E.;Liu, Leroy F.
通讯作者：
Liu, Leroy F.
DOI：
10.1038/sj.thj.6200573
发表时间：
2005-01-01
期刊：
HEMATOLOGY JOURNAL
影响因子：
--
作者：
Guerra, J;De Jesus, A;Crespo, MJ
通讯作者：
Crespo, MJ