Efentakis P, Rizakou A, Christodoulou E, Chatzianastasiou A, López M G, León R, Balafas E, Kadoglou N P E, Tseti I, Skaltsa H, Kostomitsopoulos N, Iliodromitis E K, Valsami G, Andreadou I
National and Kapodistrian University of Athens, Laboratory of Pharmacology, Faculty of Pharmacy, Athens, Greece.
National and Kapodistrian University of Athens, Laboratory of Biopharmaceutics, Faculty of Pharmacy, Athens, Greece.
Nutr Metab Cardiovasc Dis. 2017 Oct;27(10):919-929. doi: 10.1016/j.numecd.2017.08.005. Epub 2017 Aug 31.
Saffron is an antioxidant herbal derivative; however, its efficacy as a nutritional cardioprotective agent has not been fully elucidated. We investigated the cardioprotective properties of a standardized saffron aqueous extract (SFE) against ischemia/reperfusion (I/R) injury in Wild-Type (WT) and ApoE mice and the underlying molecular mechanisms.
WT and ApoE mice were subjected to 30 min I and 2 h R, with the following per os interventions for 4 weeks: 1) WT Control Group, receiving Water for Injection (WFI); 2) WT Crocus Group, receiving SFE at a dose of 60 mg/kg/day; 3) WT Crocus + Wort group, receiving SFE as described above and wortmannin at a dose of 60 μg/kg bolus 15 min before R; 4) ApoE Control Group, receiving WFI; 5) ApoE Crocus Group, receiving SFE at a dose of 60 mg/kg/day and 6) ApoE Crocus + Wort: receiving SFE as described above and wortmannin at a dose of 60 μg/kg bolus, 15 min before R. Ischemic area/area at risk (I/R%) ratio was measured. Blood samples and ischemic myocardial tissue were collected at the 10th min of reperfusion for assessment of troponin I, malondialdehyde (MDA), nitrotyrosine (NT), p-eNOS, eNOS, p-Akt, Akt, p-p42/p-p44, p-GSK3β, GSK3β, IL-6, Nrf2, HO-1 and MnSOD expression. The effect of SFE on Nrf2 expression was also evaluated in vitro. SFE reduced infarct size in WT (16.15 ± 3.7% vs 41.57 ± 2.48%, ***p < 0.001) and in ApoE mice (16.14 ± 1.47% vs 45.57 ± 1.73%, ***p < 0.001). The administration of wortmannin resulted in partial inhibition of the infarct size limitation efficacy of SFE (in both WT and Apo-E mice). Mice receiving SFE showed increased levels of eNOS, p-Akt, p-ERK1/2, p-44/p-42 and p-GSK3β-Ser9 and reduced expression of IL-6 and iNOS; furthermore, SFE reduced the levels of MDA and NT. SFE induced Nrf2 expression and its downstream targets, HO-1 and MnSOD in the myocardium of the treated animals, and induced Nrf2 expression in vitro in a dose-dependent manner.
SFE limits myocardial infarction in Wild-Type and ApoE mice in a multifaceted manner including activation of Akt/eNOS/ERK1/2/GSK3-β and through Nrf2 pathway, bestowing antioxidant protection against I/R.
藏红花是一种抗氧化草本提取物；然而，其作为营养性心脏保护剂的功效尚未完全阐明。我们研究了标准化藏红花水提取物（SFE）对野生型（WT）和载脂蛋白E（ApoE）小鼠缺血/再灌注（I/R）损伤的心脏保护特性及其潜在分子机制。
WT和ApoE小鼠经历30分钟缺血和2小时再灌注，并进行以下经口干预4周：1）WT对照组，接受注射用水（WFI）；2）WT藏红花组，接受剂量为60mg/kg/天的SFE；3）WT藏红花+渥曼青霉素组，接受上述SFE，并在再灌注前15分钟接受剂量为60μg/kg的渥曼青霉素推注；4）ApoE对照组，接受WFI；5）ApoE藏红花组，接受剂量为60mg/kg/天的SFE；6）ApoE藏红花+渥曼青霉素组：接受上述SFE，并在再灌注前15分钟接受剂量为60μg/kg的渥曼青霉素推注。测量缺血面积/危险面积（I/R%）比值。在再灌注第10分钟采集血样和缺血心肌组织，以评估肌钙蛋白I、丙二醛（MDA）、硝基酪氨酸（NT）、磷酸化内皮型一氧化氮合酶（p-eNOS）、内皮型一氧化氮合酶（eNOS）、磷酸化蛋白激酶B（p-Akt）、蛋白激酶B（Akt）、磷酸化细胞外信号调节激酶1/2（p-p42/p-p44）、磷酸化糖原合成酶激酶3β（p-GSK3β）、糖原合成酶激酶3β（GSK3β）、白细胞介素-6（IL-6）、核因子E2相关因子2（Nrf2）、血红素加氧酶-1（HO-1）和锰超氧化物歧化酶（MnSOD）的表达。还在体外评估了SFE对Nrf2表达的影响。SFE减小了WT小鼠（16.15±3.7%对41.57±2.48%，***p<0.001）和ApoE小鼠（16.14±1.47%对45.57±1.73%，***p<0.001）的梗死面积。渥曼青霉素的给药导致SFE对梗死面积限制功效的部分抑制（在WT和Apo-E小鼠中均如此）。接受SFE的小鼠显示eNOS、p-Akt、p-ERK1/2、p-44/p-42和p-GSK3β-Ser9水平升高，IL-6和诱导型一氧化氮合酶（iNOS）表达降低；此外，SFE降低了MDA和NT水平。SFE诱导了治疗动物心肌中Nrf2表达及其下游靶点HO-1和MnSOD，并在体外以剂量依赖性方式诱导Nrf2表达。
SFE以多方面的方式限制野生型和ApoE小鼠的心肌梗死，包括激活Akt/eNOS/ERK1/2/GSK3-β并通过Nrf2途径，赋予对I/R的抗氧化保护作用。
