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目的体外探讨马尾松树皮提取物(Pinus massoniana Bark Extracts,PMBE)对乙醇损伤人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVEC)的保护作用及PMBE是否通过调控生长分化因子15(Growth Differentiation Factors 15,GDF-15)的表达从而发挥其作用。方法 60μg/ml的PMBE与HUVEC共培养12 h后加入4%(V/V)浓度的乙醇作用12 h进行各项研究:MTT法检测细胞存活率;观察细胞形态;Annexin V-FITC/PI双染法检测细胞凋亡率;分别检测细胞内ROS水平、超氧化物含量及SOD活性,细胞培养液上清中的NO含量;RT-PCR检测细胞内GDF-15的mRNA表达及Western Blot检测细胞内GDF-15的蛋白表达情况。结果与4%乙醇组相比,PMBE能明显提高细胞存活率,降低细胞早期凋亡和坏死比例,ROS水平及超氧化物含量降低,SOD活性及培养液上清中NO含量升高,GDF-15的mRNA表达水平降低,GDF-15的蛋白表达水平降低。结论 PMBE对乙醇损伤HUVEC具有保护作用,其机制可能与影响GDF-15的表达水平相关。
Objective To investigate the protective effect of Pinus massoniana Bark Extracts (PMBE) on ethanol-injured human umbilical vein endothelial cells (HUVECs) in vitro and the role of PMBE in the regulation of Growth Differentiation Factors 15 , GDF-15) to play its role. Methods PMBE (60μg / ml) was co-cultured with HUVEC for 12 hours and then treated with 4% (V / V) ethanol for 12 hours. Cell viability was measured by MTT assay. Cell morphology was observed by Annexin V-FITC / The cell apoptosis rate was detected by flow cytometry. The intracellular ROS levels, superoxide and superoxide dismutase (SOD) activity and NO content in cell culture supernatant were detected respectively. The mRNA expression of GDF-15 in cells was detected by RT-PCR and Western Blot GDF-15 in the protein expression. Results Compared with 4% ethanol group, PMBE could significantly improve the cell viability, reduce the proportion of early apoptosis and necrosis, reduce the levels of ROS and superoxide dismutase, increase the activity of SOD and the content of NO in the supernatant, 15 mRNA expression decreased, GDF-15 protein expression decreased. Conclusion PMBE has a protective effect on HUVEC induced by ethanol, and its mechanism may be related to the expression of GDF-15.