目的 探讨谷红注射液对乳鼠脑微血管内皮细胞因缺糖缺氧所致损伤的保护作用。方法 建立乳鼠脑微血管内皮细胞缺糖缺氧损伤模型。利用四甲基偶氮唑蓝法和预实验确定加药浓度后,将培养的细胞随机分成6组:正常组、模型组、谷红注射液低、中、高治疗组(20、60、100 μLmL-1)和阳性药物对照组(尼莫地平,200 μgmL-1)。观察谷红注射液对脑微血管内皮细胞一氧化氮(NO)和丙二醛(MDA)含量、超氧化歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)活性的影响;并采用流式细胞仪检测各组细胞的凋亡率和细胞周期变化。结果 与模型组相比,谷红注射液组特别是高剂量组能显著降低胞内一氧化氮、丙二醛含量,显著增强胞内超氧化物歧化酶、谷胱甘肽过氧化物酶活性(P＜0.01);谷红注射液中、高剂量组可明显改善脑微血管内皮细胞形态,抑制细胞凋亡,有效抑制缺糖缺氧诱导的细胞发生G1/S期阻滞。结论 谷红注射液对乳鼠脑微血管内皮细胞的缺糖缺氧损伤显示出良好的保护作用,其机制可能与抗氧化、抑制细胞凋亡的作用有关。
Abstract
OBJECTIVE To investigate the protective effect of Guhong injection(GH) on rat cerebral microvascular endothelial cells (rBMECs) injured by oxygen-glucose deprivation. METHODS The model of oxygen-glucose deprivation in rBMECs was established. After the concentrations of drug were selected by the MTT method and pre-experiment, the cultured cells were randomly divided into six groups: control group; vehicle group; different GH dosage groups (20,60,100 μLmL-1); positive control group (nimodipine, 200 μgmL-1). The effects of GH on NO level, malondialdehyde (MDA) level, superoxide dismutase (SOD) activity and glutathione peroxidase (GSH-PX) activity were evaluated. The apoptosis rate and cell cycle were determined by flow cytometry. RESULTS Compared with model group, GH-treated groups especially the high dose group can remarkablely reduce the content of intracellular NO and MDA, and enhance the activity of SOD and GSH-PX (P＜0.01). The morphology of rBMECs was improved obviously, the number of apoptosis cells induced by hypoxia was reduced significantly, and the blockage of rBMECs G1/S phase induced by oxygen-glucose deprivation was effectively inhibited in the middle and high dose groups. CONCLUSION GH exhibits a strong ameliorative protection when rBMECs presented with oxygen-glucose deprivation, in which the mechanism may be related to the enhancement of cellular antioxidant capacity and the inhibition of cell apoptosis.
关键词
谷红注射液 /
脑微血管内皮细胞 /
缺糖缺氧损伤 /
细胞凋亡
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Key words
Guhong injection /
cerebral microvascular endothelial cells /
oxygen-glucose deprivation /
cell apoptosis
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中图分类号：
R965
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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(81373898,81173647,81274176);浙江省自然科学基金(LR12H27001,LZ14H27001);浙江省卫生高层次创新人才培养工程项目;浙江省中医药(中西医结合)重点学科(2012-XK-A06)
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