H2O2诱导内皮细胞凋亡及其机制探讨

Study on H2O2-induced Endothelial Cell Apoptosis and Its Mechanism

【目的】探讨过氧化氢（H2O2）所致内皮细胞功能损害中的作用及其机制。【方法】人脐静脉内皮细胞（HUVEC）培养后，分为对照组（A组），H2O2处理组（B组），H2O2和蛋白激酶C（PKC）抑制剂GF109203处理组（C组），H2O2和P38抑制剂Apigenin处理组（D组）。H2O2处理细胞30min后加入相应试剂作用24h。MTT法测定细胞存活率，Ki-67染色检测HUVEC增殖活性，免疫荧光染色方法检测细胞凋亡，Western blotting检测p53蛋白表达。【结果】①B组可诱导HUVEC功能损伤，使HUVEC的存活率降低、增殖活性降低、细胞凋亡增加，并使p53蛋白的表达上调；②C组可显著抑制H20z诱导的HuVEC功能损伤和p53蛋白表达的上调；而D组不能抑制H2O2诱导的HUVEC功能损伤和p53蛋白表达的上调。【结论】p53在氧化应激所致内皮细胞功能损害中起重要作用；而这些作用与PKC信号转导通路有关。

[Objective]To explore the hydrogen peroxide（H2O2 ）-induced endothelial cell dysfunction and its mechanism. [Methods] The cultured human umbilical vein endothelial cells（HUVEC） were divided into control group（group A）, H2O2-treated group（group B）, H2O2 plus protein kinase C（PKC） inhibitor GF109203-treated group（group C） and H2O2 plus p3S inhibitor Apigenin-treated group. The cedis were treated with H2O2 for 30min, and then the reagent was added for 24h. MTT assay was used to detect the cell survival rate and Ki-67 staining was used to detect HUVEC proliferating activity. Immune fluorescent staining method was used to detect the cell apoptosis and Western blotting was used to detect the expression of p53 protein. [Results] H2O2 could in- duce HUVEC dysfunction, and decrease the survival rate and proliferating activity of HUVEC, and increase the cell apoptosis, and up-regulate the expression of p53 protein. PKC inhibitor GF109203 could markedly inhibit H2O2-induced HUVEC dysfunction and the upregulation of the expression of p53 protein, but p38 inhibitor Apigenin could not inhibit H2O2-induced HUVEC dysfunction and the upregulation of the expression of p53 protein. [Conclusion] p53 plays an important role in the endothelial dysfunction induced by oxidative stress, and the action may be related to PKC signal transduction pathway.