Division of Nephrology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan.
Division of Urology, Department of Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan.
Biomed Pharmacother. 2019 Jan;109:658-670. doi: 10.1016/j.biopha.2018.10.095. Epub 2018 Nov 4.
This study tested the hypothesis that early administration of empagliflozin (Empa), an inhibitor of glucose recycling in renal tubules, could preserve heart function in cardiorenal syndrome (CRS) in rat. Chronic kidney disease (CKD) was caused by 5/6 subtotal nephrectomy and dilated cardiomyopathy (DCM) by doxorubicin (DOX) treatment. In vitro results showed that protein expressions of cleaved-caspase3 and autophagy activity at 24 h/48 h in NRK-52P cells were significantly upregulated by para-Creso treatment; these were significantly downregulated by Empa treatment. Flow cytometric analysis showed that annexin-V (i.e., early/late apoptosis) in NRK-52P cells expressed an identical pattern to cleaved-caspase3 between the two groups (all p < 0.001). Adult-male-SD rats (n = 18) were equally categorized into group 1 (sham-control), group 2 (CRS) and group 3 [CRS + Empa; 20 mg/kg/day]. By day-42 after CRS induction, left-ventricular ejection fraction (LVEF) level exhibited an opposite pattern, whereas LV end-diastolic dimension and creatinine level displayed the same pattern, to cleaved-caspase3 among the three groups (all p < 0.0001). In LV tissues, protein expressions of inflammatory (tumor-necrosis factor-α/nuclear-factor-κB/interleukin-1ß/matrix-metalloprotianse-9), oxidative stress (NOX-1/NOX-2/oxidized protein), apoptotic (mitochondrial-Bax/cleaved-caspase-3/cleaved-PARP), fibrotic (transforming-growth factor-ß/Smad3), DNA/mitochondrial-damage (γ-H2AX/cytosolic-cytochrome-C) and heart failure (brain natriuretic peptide (BNP) levels displayed an opposite pattern to LVEF among the three groups (all p < 0.0001). Additionally, cellular expressions of DNA-damage/heart-failure (γ-H2AX+//XRCC1+CD90+//BNP+) biomarkers and histopathological findings of fibrotic/condensed collagen-deposition areas and apoptotic nuclei showed an identical pattern, whereas connexin43 and small-vessel number exhibited an opposite pattern, to inflammation among the three groups (all p < 0.0001). In conclusion, Empa therapy protected heart and kidney against CRS injury.
这项研究检验了这样一个假设，即早期给予肾小管葡萄糖再循环抑制剂恩格列净（empagliflozin）可预防 5/6 肾切除和阿霉素（doxorubicin，DOX）治疗诱导的大鼠心肾综合征（cardiorenal syndrome，CRS）中的心脏功能障碍。慢性肾脏病（chronic kidney disease，CKD）由 5/6 肾切除引起，扩张型心肌病（dilated cardiomyopathy，DCM）由 DOX 治疗引起。体外结果表明，para-Creso 处理可使 NRK-52P 细胞中 caspase3 剪切和自噬活性的蛋白表达在 24 小时/48 小时显著上调；而 Empa 处理则显著下调。流式细胞术分析显示，NRK-52P 细胞中的膜联蛋白-V（即早期/晚期凋亡）与两组之间的 caspase3 表达呈相同模式（均 p<0.001）。18 只成年雄性 SD 大鼠（n=18）等分为 3 组：第 1 组（sham-control）、第 2 组（CRS）和第 3 组[CRS+Empa；20mg/kg/天]。在 CRS 诱导后第 42 天，左心室射血分数（left-ventricular ejection fraction，LVEF）水平在三组中呈现相反的模式，而左心室舒张末期内径和肌酐水平在三组中呈现相同的模式，均与 caspase3 呈正相关（均 p<0.0001）。在左心室组织中，炎症（肿瘤坏死因子-α/核因子-κB/白细胞介素-1β/基质金属蛋白酶-9）、氧化应激（NOX-1/NOX-2/氧化蛋白）、凋亡（线粒体-Bax/caspase3 剪切体/caspase-3 剪切体/cleaved-PARP）、纤维化（转化生长因子-β/Smad3）、DNA/线粒体损伤（γ-H2AX/细胞质细胞色素 C）和心力衰竭（脑钠肽（brain natriuretic peptide，BNP）水平的蛋白表达与三组中 LVEF 的变化模式相反（均 p<0.0001）。此外，细胞 DNA 损伤/心力衰竭（γ-H2AX+/+/XRCC1+/CD90+/BNP+）生物标志物的表达和纤维化/浓缩胶原沉积区和凋亡核的组织病理学发现与炎症呈相同模式，而连接蛋白 43 和小血管数量与炎症呈相反模式，与炎症呈相同模式（均 p<0.0001）。总之，Empa 治疗可预防 CRS 损伤导致的心肾损伤。
