黄连素调节eNOS/NO保护软脂酸诱导的HUVECs损伤作用机制研究

Protective Effects of Berberine on Palmitate-Induced Injury in HUVECs by Regulating e NOS/NO

目的研究黄连素对软脂酸诱导人脐静脉内皮细胞(HUVECs)损伤的保护作用,并初步探讨其作用机制.方法采用500μmol/L软脂酸培养HUVECs 24 h,建立内皮细胞损伤模型,MTT法观察黄连素对细胞存活率的影响;检测细胞培养上清液中一氧化氮(NO)含量;RT-PCR法检测内皮型一氧化氮合酶(e NOS)mRNA水平;Western blot方法检测e NOS和磷酸化e NOS蛋白的表达.结果与对照组比较,软脂酸组细胞存活率降低(P<0.05),培养液中NO含量下降(P<0.05),细胞内e NOS mRNA水平、e NOS及磷酸化e NOS蛋白表达水平均显著下降(P<0.01,P<0.05).与软脂酸组比较,黄连素组细胞存活率增加,培养液中NO含量明显提高(P<0.05),细胞内e NOS mRNA水平和磷酸化蛋白表达水平均显著提高(P<0.01,P<0.05).结论黄连素对软脂酸引起的血管内皮细胞损伤有显著的保护作用,并可能与上调e NOS、促进NO生成有关.

Objective To explore the protective effect of berberine on palmitate-induced injury in human umbilical vein endothelial cells （ HUVECs ） and reveal its mechanism. Method The endothelial cell injury model was established by culturing HUVECs with 500 μmol/L palmitate for 24 h. The cell viability of HUVECs was observed by MTT assays. The content of Nitric oxide （ NO ） in the supernatant was determined. The mRNA level of endothelial nitric oxide synthase（eNOS）was measured by RT-PCR,and the protein levels of eNOS and p-eNOS were analyzed by western blot. Results Compared with those in the control group,the cell viability of HUVECs, the content of NO in supernatant,mRNA level of eNOS,and the protein levels of eNOS and p-eNOS in palmitate group were significantly decreased（P〈0. 01,P〈0. 05）. Compared with that in the palmitate group,the cell viability of HUVECs in the berberine group was increased. NO content,mRNA level of eNOS,and the protein levels of eNOS and p-eNOS in the berberine group were all significantly increased（P〈0. 01 or P〈0. 05）. Conclusion Berberine has a significant protective effect on palmitate-induced endothelial cell injury,which might be related to the up-regulation of eNOS and the promotion of NO production.