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观察不同浓度同型半胱氨酸(homocysteine,HCY)在Cu2+介导下,能否诱导培养的人脐静脉内皮细胞凋亡,以揭示HCY致血管内皮损伤的机制。采用细胞计数板检测脱落细胞量;比色法分别测定乳酸脱氢酶释放率、细胞内丙二醛含量和超氧化物歧化酶、谷胱甘肽过氧化物酶活力的改变;Hoechst 33258染色观察凋亡细胞核形态变化及流式细胞术定量测定细胞凋亡。结果表明HCY在生理浓度Cu2+的介导下,可能通过氧化应激损伤的机制而导致血管内皮细胞凋亡,这提示在体内可能通过此途径诱发动脉粥样硬化。
To observe whether different concentrations of homocysteine (HCY) induce the apoptosis of cultured human umbilical vein endothelial cells under the condition of Cu2 +, so as to reveal the mechanism of HCY induced vascular endothelial injury. The amount of exfoliated cells was detected by using cell counting plate. The release of lactate dehydrogenase (LDH), intracellular malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (POD) activity were determined by colorimetry. Hoechst 33258 staining Morphological changes of apoptotic nuclei and flow cytometry were used to quantitatively determine apoptosis. The results showed that under the physiological concentration of Cu2 +, HCY may induce the apoptosis of vascular endothelial cells through the mechanism of oxidative stress injury, which suggests that atherosclerosis may be induced by this pathway in vivo.