目的 探讨祁州漏芦水提取物(Radix Rhapontici water extract,RRWE)对叔丁基过氧化氢(tert-butyl hydroperoxide,TBHP)所致血管内皮细胞凋亡的保护作用。方法 体外利用TBHP制备人脐静脉血管内皮细胞损伤模型,随机分为正常对照组、TBHP损伤组、RRWE低剂量组和高剂量组。采用四甲基偶氮唑蓝(MTT)法检测细胞活力,比色法观察丙二醛(MDA)、还原型谷胱甘肽(GSH)含量和超氧化物歧化酶(SOD)活性,采用荧光染色法观察活性氧(ROS)、细胞凋亡及线粒体跨膜电位水平,免疫印迹法检测细胞NF-κB、JNK、Bcl-2、Bax和caspase-3蛋白的表达情况。结果 RRWE增高TBHP所致损伤的内皮细胞活力,降低细胞ROS水平,减少MDA生成,增高GSH水平与SOD活性,升高线粒体跨膜电位水平,并降低细胞p-JNK及p-NF-κB水平,降低细胞Bax/Bcl-2比值,减少caspase-3活化,抑制细胞凋亡。结论 RRWE对TBHP所致的血管内皮细胞损伤具有保护作用,并抑制细胞凋亡,其机制可能与抑制JNK和NF-κB活化有关。
Abstract
OBJECTIVE To explore the protective effect of Radix Rhapontici water extract (RRWE) on the damage of vascular endothelial cells induced by tert-butyl hydroperoxide (TBHP) in vitro. METHODS The cellular model was established by treating human umbilical vein endothelial cells with TBHP, and randomly assigned to 4 groups:the control, TBHP, low and high-dose RRWE groups. Cell viability was tested by MTT assay, the levels of reduced glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) were measured by colorimetric method, the reactive oxygen species (ROS), apoptosis, and mitochondrial membrane potentials were observed by fluorescent staining, and the protein expressions of NF-κB, JNK, Bax, Bcl-2 and caspase-3 were determined with Western blotting method. RESULTS Pretreatment with RRWE significantly increased the cell viabilities, reduced ROS levels, decreased MDA formation, increased the GSH contents and SOD activities, elevated the mitochondrial membrane potentials, down-regulated the p-JNK and p-NF-κB levels, reduced Bax/Bcl-2 ratios, suppressed caspase-3 activation, and inhibited cell apoptosis of vascular endothelial cells. CONCLUSION RRWE has a protective effect on the damage of vascular endothelial cells induced by TBHP in vitro, and suppresses the cell apoptosis maybe through inhibiting JNK and NF-κB activation.
关键词
祁州漏芦 /
水提取物 /
血管内皮细胞 /
叔丁基过氧化氢 /
凋亡
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Key words
Radix Rhapontici /
water extract /
vascular endothelial cells /
tert-butylhydroperoxide /
apoptosis
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中图分类号：
R285.5
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脚注
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基金
国家自然科学基金项目资助(81160539)
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