摘要
目的 观察丹参酮ⅡA磺酸钠(sodium tanshinone ⅡA sulfonate， STS)对血管紧张素Ⅱ(angiotensin Ⅱ， Ang Ⅱ)诱导的心肌细胞氧化应激反应的影响及其机制。方法 培养新生大鼠心肌细胞，建立Ang Ⅱ诱导的心肌细胞氧化应激反应模型。用活性氧(reactive oxygen species， ROS)敏感的二氮荧光黄双乙酸盐（DCFH-DA）探针测定胞内ROS水平。酶联免疫分析法测定细胞培养物上清中8-羟脱氧鸟苷（8-OHdG）含量。以细胞存活率、细胞内超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量、培养上清液乳酸脱氢酶(LDH)活性作为反应心肌细胞损伤的指标。比色法测定还原型辅酶Ⅱ(NADPH)氧化酶(NADPH oxidase， NOX)活性。蛋白质印迹法（Western-blot）测定NOX亚单位p47 phox表达。结果 Ang Ⅱ能显著提高心肌细胞内DCF荧光信号强度，降低细胞存活率及SOD活性，同时上调培养上清液LDH活性及MDA含量。以上氧化应激相关指标能被各浓度STS抑制。进一步研究发现，STS能抑制Ang Ⅱ上调的NADPH氧化酶活性及亚单位p47 phox表达。结论 STS可以抑制Ang Ⅱ诱导的心肌细胞氧化应激损伤反应，机制可能与下调NADPH氧化酶活性及亚单位p47 phox表达有关。
Abstract
OBJECTIVE To observe the effects and mechanisms of sodium tanshinone ⅡA sulfonate (STS) on angiotensin Ⅱ (Ang Ⅱ)-induced cardiomyocyte oxidative stress. METHODS In the primary culture of neonatal rat cardiomyocytes， the content of reactive oxygen species (ROS) was measured by 2，7-dichlorofluorescein diacetate (DCFH-DA). 8-hydroxydeoxyguanosine level in the supernatant was measured by ELISA. As indexes of cardiomyocyte oxidative stress， the cellular contents of MDA and SOD， cell vialibity and LDH release were measured. NADPH oxidase (NOX) activity was measured by chromatometry. The expressions of p47 phox was assessed using Western blot. RESULTS STS can decrease Ang Ⅱ-induced elevations of ROS level and oxidative stress， and inhibit the expression of p47 phox and NOX activity. CONCLUSION The inhibitory effects of STS on Ang Ⅱ-induced cardiomyocyte oxidative stress may be associated with depressing NOX signaling pathway via down regulation of p47 phox expression.
关键词
丹参酮ⅡA磺酸钠 /
心肌细胞肥大 /
活性氧 /
NADPH氧化酶
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Key words
sodium tanshinone ⅡA sulfonate /
cardiomyocyte hypertrophy /
reactive oxygen species /
NADPH oxidase
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杨乐 邹晓静 高翔 李树生 梁黔生 杨光田.
丹参酮ⅡA磺酸钠对Ang Ⅱ诱导的心肌细胞氧化应激的影响[J]. 中国药学杂志, 2012, 47(4): 270-273
Effects of Sodium Tanshinone ⅡA Sulfonate on Ang Ⅱ-Induced Cardiomyocyte Oxidative Stress[J]. Chinese Pharmaceutical Journal, 2012, 47(4): 270-273
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参考文献
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脚注
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基金
国家自然科学基金资助项目（81102691；30901405）；湖北省自然科学基金（2011CDB538）
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