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目的:观察三七总皂苷(PNS)对过氧化氢损伤人脐静脉内皮细胞(HUVECs)LDH释放量、胞内SOD活性的影响,研究其对受损细胞粘附分子ICAM-1及MCP-1 mRNA表达的影响,探讨三七总皂苷对HUVECs的保护作用及其在动脉粥样硬化中作用的可能机制。方法:三七总皂苷预孵育细胞4h后,加入300μmol/L H2O2,继续培养24h,MTT法检测细胞活力,采用试剂盒测定LDH释放量、SOD酶活力,荧光实时定量PCR(RT-PCR)法检测三七总皂苷作用下内皮细胞ICAM-1及MCP-1 mRNA的表达。结果:三七总皂苷在25~100μg/ml浓度范围内可显著提高受损细胞活力,并能显著降低LDH漏出率,提高胞内SOD酶活力;RT-PCR结果表明H2O2组内皮细胞表达的ICAM-1和MCP-1 mRNA明显高于正常细胞组,而同时加入50及100μg/ml的三七总皂苷后两者mRNA表达明显下降。结论:三七总皂苷可保护血管内皮细胞免受H2O2的损害,改善受损细胞氧化指标,降低参与动脉粥样硬化相关细胞因子表达,从而预防和治疗动脉粥样硬化。
Objective: To observe the effect of Panax Notoginseng Saponins (PNS) on the release of LDH and the activity of SOD in H2O2 injured human umbilical vein endothelial cells (HUVECs), and to study the effect of PNS on the expression of ICAM-1 and MCP-1 To investigate the protective effect of Panax notoginseng saponins on HUVECs and its possible mechanism in atherosclerosis. Methods: The cells were pre-incubated with total saponins of Panax notoginseng for 4 hours, and then incubated with 300μmol / L H2O2 for 24 hours. Cell viability was measured by MTT assay. LDH release, SOD activity and real-time quantitative PCR (RT-PCR) To detect the expression of ICAM-1 and MCP-1 mRNA in endothelial cells under the action of Panax notoginseng saponins. Results: The total saponins of Panax notoginseng at 25 ~ 100μg / ml concentration could significantly increase the viability of damaged cells, and can significantly reduce the leakage rate of LDH and increase intracellular SOD activity; RT-PCR results showed that H2O2 group of endothelial cells expressed ICAM -1 and MCP-1 mRNA were significantly higher than those in normal cells. However, the mRNA expression of both Panax notoginseng saponins in 50 and 100 μg / ml groups decreased significantly. Conclusion: Panax notoginseng saponins can protect vascular endothelial cells from H2O2 damage, improve the oxidative index of damaged cells and reduce the expression of cytokines involved in atherosclerosis, thus preventing and treating atherosclerosis.