8-硝基白杨素对溶血性磷脂酰胆碱损伤内皮细胞的保护作用

Protective Effect of 5,7-Dihydroxyl-8-nitrio Chrysin on the Growth of Vascular Endothelial Cell Damaged by Lysophosphatidyl Choline

目的研究8-硝基白杨素(8-NOChR)对溶血性磷脂酰胆碱(LPC)损伤内皮细胞的保护作用及可能机制。方法体外培养人脐静脉内皮细胞(HUVEC),MTT检测LPC对HUVEC的损伤作用及不同浓度8-NOChR拮抗LPC损伤HUVEC生长作用;平皿克隆检测不同浓度的8-NOChR拮抗LPC抑制HUVEC细胞克隆形成;FCM检测8-NOChR调整LPC损伤HUVEC细胞周期变化;用Western blot检测细胞周期相关蛋白表达。结果 MTT提示经LPC作用的HUVEC生长活性呈时间和浓度依赖性降低,与NS组或0.1%DMSO组比较差异有统计学意义(P<0.05);与LPC组相比较,8-NOCHR呈浓度依赖性地提高LPC损伤的HUVEC生长活性(P<0.05),提高细胞克隆形成率(P<0.05);降低HUVEC G2/M期数目(P<005);同时Cyclin A和Cyclin B蛋白激活(P<0.05),P53和P21蛋白表达下调(P<0.05),而CDK1蛋白表达却不变(P>0.05)。结论 8-NOCHR拮抗LPC损伤的内皮细胞的生长活性,可能与其激活Cyclin A和Cyclin B,下调P53和P21蛋白表达相关。

Aim To investigate the protective effect of 5,7-Dihydroxyl-8-nitrio chrysin （8-NOChR） on the growth of vascular endothelial cell damaged by lysophosphatidyl choline （ LPC ） and its possible molecular mechanism. Methods Human umbilical vein endothelial cells （HUVEC） were incubated in vitro, the damaged effect of LPC on HU- VEC cells and the rivalry effect of various concentration of 8-NOChR on HUVEC cells induced by LPC were evaluated by MTT assay. The colony formations were detected by plate colony formation assay. Distribution of cell cycle was analyzed by flow cytometry. Western blot assay was used to measure proteins related to cell cycle. Results MTT showed the viability of HUVEC treated with LPC decreased in a dose and time dependent manner compared with the cells treated with NS group and 0. 1% DMSO group （ P 〈 0. 05 ）, while 8-NOChR could increase the viability and the colony formations of eells treated with LPC in a dose and time-dependent manner compared with the LPC group （ P 〈 0. 05 ）. FCM indicated the G2/M accumulation of HUVEC treated with LPC combining 8-NOChR decreased in dose-dependent, and the expression of eyclin A and cyclin B protein were activated （P 〈0. 05） , and protein level of P53 and P21 were down-regulated （P 〈 0. 05 ）, while the expression of CDK1 protein showed no change with the same treatment （ P 〉 0.05 ）. Conclusion 8- NOChR could significantly prevent the damaged HUVEC induced by LPC, which correlated with activation of eyclin A and cyclin B protein and down-regulation of P53 and P21 protein.