Zhang S M, Xie Z P, Xu M L, Shi L F
School of Pharmacy, Binzhou Medical College , Yantai , China and.
Pharm Biol. 2015;53(9):1352-7. doi: 10.3109/13880209.2014.982298. Epub 2015 Apr 15.
Cardiomyocyte apoptosis plays a critical role in the progress of heart diseases. Fucoidan, a complex-sulfated polysaccharide, has been reported to possess potential cardioprotective efficacy in vivo.
The present study determines whether fucoidan could provide cardioprotection on hypoxia-induced cardiomyocyte apoptosis.
H9c2 cardiomyoblast cells were incubated with various concentrations (15, 30, and 60 μg/ml) of fucoidan in a humidified incubator at 37 °C with 95% O2 and 5% CO2. After 6 h, hypoxia was processed and the cardioprotective effects of fucoidan were evaluated by applying MTT, ELISA, Hoechst 33258 nucleus staining, and western blot.
Following a 6 h exposure of H9c2 to hypoxic condition, significant reduction was found in cell survival (0.57-fold) and superoxide dismutase (SOD) activity (0.56-fold), which were associated with the increase of malondialdehyde (MDA) level (2.58-fold), creatine phosphokinase (CK, 3.57-fold), and lactate dehydrogenase (LDH) activities (2.39-fold). Moreover, hypoxia-induced apoptosis was confirmed by Hoechst 33258 nuclear staining, and these changes were accompanied by the increase of Bcl-2 (1.27-fold) and Bax expression (2.6-fold). However, preincubation of the cells with fucoidan prior to hypoxia exposure elevated the cell viability (30 μg/ml, 1.18-fold; 60 μg/ml, 1.32-fold) and SOD activity (30 μg/ml, 1.12-fold; 60 μg/ml, 1.25-fold), but decreased the MDA level (30 μg/ml, 0.70-fold; 60 μg/ml, 0.80-fold), CK (30 μg/ml, 0.69-fold; 60 μg/ml, 0.76-fold), and LDH (30 μg/ml, 0.67-fold; 60 μg/ml, 0.86-fold) leakages. Hoechst 33258 nuclear staining observations demonstrated the same protective effect of fucoidan on hypoxia-induced myocardial injury. Also, cardioprotective effects of fucoidan were reflected by increasing Bcl-2 (60 μg/ml, 1.84-fold), as well as decreasing Bax (60 μg/ml, 0.6-fold).
Fucoidan had protective effect against hypoxia-induced cardiomyocytes apoptosis, and the mechanism might involve protections of the cell from oxidative injury.
心肌细胞凋亡在心脏病进展中起关键作用。岩藻依聚糖是一种复合硫酸化多糖，据报道在体内具有潜在的心脏保护功效。
本研究确定岩藻依聚糖是否能对缺氧诱导的心肌细胞凋亡提供心脏保护作用。
将H9c2心肌母细胞在37℃、含95%氧气和5%二氧化碳的湿度培养箱中，用不同浓度（15、30和60μg/ml）的岩藻依聚糖孵育。6小时后，进行缺氧处理，并通过MTT、ELISA、Hoechst 33258细胞核染色和蛋白质印迹法评估岩藻依聚糖的心脏保护作用。
H9c2细胞暴露于缺氧条件6小时后，细胞存活率（0.57倍）和超氧化物歧化酶（SOD）活性（0.56倍）显著降低，这与丙二醛（MDA）水平升高（2.58倍）、肌酸磷酸激酶（CK，3.57倍）和乳酸脱氢酶（LDH）活性升高（2.39倍）相关。此外，Hoechst 33258细胞核染色证实了缺氧诱导的凋亡，并且这些变化伴随着Bcl-2表达增加（1.27倍）和Bax表达增加（2.6倍）。然而，在缺氧暴露前用岩藻依聚糖对细胞进行预孵育可提高细胞活力（30μg/ml，1.18倍；60μg/ml，1.32倍）和SOD活性（30μg/ml，1.12倍；60μg/ml，1.25倍），但降低了MDA水平（30μg/ml，0.70倍；60μg/ml，0.80倍）、CK（30μg/ml，0.69倍；60μg/ml，0.76倍）和LDH泄漏（30μg/ml，0.67倍；60μg/ml，0.
