[益生注射液对内皮细胞缺氧-再氧化损伤的保护机制研究]。

Shengfu Li, Hongmei Jiang, Youping Li, Darong Wu, Li Zhang, Lin Zhang, Jingqiu Cheng
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引用次数: 0

摘要

目的:探讨缺血再灌注损伤(IRI)的机制及益生注射液对内皮细胞(ECs)的保护作用。方法:将人脐静脉内皮细胞系ECV304在不含葡萄糖的RPMI培养基1640中100% N2下培养一个半小时,然后在正常条件下含葡萄糖的RPMI培养基1640中培养5小时,模拟ECs移植后的IRI。实验组在培养基中加入纯天然药物益生注射液。荧光染色显示细胞内钙和线粒体。细胞化学染色法检测琥珀酸脱氢酶(SDH)和乳酸脱氢酶(LDH)活性。测定培养基中超氧化物歧化酶(SOD)、丙二醛(MDA)和一氧化二氮(NO)的浓度。结果:缺氧-复氧处理后,内皮细胞形态学改变,细胞内钙浓度、LDH活性、MDA浓度升高。线粒体膜电位降低,SOD、NO浓度降低。益生注射液使这些变化消失或减少。结论:缺氧-再氧化可引起内皮细胞脂质过氧化、钙超载及线粒体功能损伤。益生注射液可以逆转这些变化,从而保护内皮细胞。
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[A study on the protective mechanism of yisheng injection against the anoxia-reoxygenation injury to endothelial cells].

Objective: To explore the mechanism of ischemia reperfusion injury (IRI) and the protective effect of Yisheng injection on endothelial cells (ECs).

Methods: Human umbilical veins endothelial cell line ECV304 was cultured in RPMI medium 1640 without glucose under 100% N2 for one and a half hours, and then in RPMI medium 1640 with glucose under normal conditions for 5 hours to mimic ECs' IRI posttransplantaion. A pure nature medicine, Yisheng injection was added into the medium in experimental group. Intracellular calcium and mitochondrial were shown through flourescent staining. The activities of succinic dehydrogenase (SDH) and lactic dehydrogenase (LDH) were detected by cytochemical staining. The concentrations of superoxide dismutase (SOD), malonaldehyde (MDA) and nitrous oxide (NO) in culture medium were determined.

Results: After treatment with anoxia-reoxygenation, the ECs' morphologic changes were observed, and intracellular calcium concentration, LDH activity, MDA concentration became higher. Meanwhile, mitochondrial membrane potential, concentrations of SOD and NO were lower. Yisheng injection made these changes disappear or become less.

Conclusion: Anoxia-reoxygenation induces lipid peroxidation, calcium superrcharge in ECs, and damage to mitochondrial function. Yisheng injection can reverse these changes, thus protecting the endothelial cells.

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