目的 通过检测姜黄素脂质纳米粒对人脐静脉内皮细胞(HUVEC)分泌一氧化氮(NO)与细胞间粘附分子(ICAM-1)表达水平的影响,探讨新剂型对于动脉粥样硬化疾病的抑制作用.方法 制备姜黄素脂质纳米粒,培养HUVEC细胞,随机分正常组、模型组、脂质纳米粒空载药组,三组剂量呈梯度的姜黄素脂质纳米粒药物组.10μg/LTNF-α刺激后,加入5～60mg/L姜黄素纳米制剂,用MTT增殖实验检测姜黄素脂质纳米粒对HUVEC细胞增殖作用的影响;采用实时荧光定量PCR测定一氧化氮合酶(eNOS)与ICAM-1的基因表达水平;通过ELISA法观察ICAM-1的含量;用硝酸还原酶法检测NO表达量.结果 MTT法检测显示5～30mg/L姜黄素脂质纳米粒对细胞活力无明显影响,当药物浓度为60mg/L时,活细胞数量有所减少.与正常组比较,模型组eN0S表达量降低(P<0.01),姜黄素脂质纳米粒预处理的HUVEC细胞可显著上调eNOS基因表达(P<0.01),促进NO分泌量增多(P<0.01);TNF-α引起的血管细胞粘附分子ICAM-1中mRNA的表达上升(P<0.01);姜黄素脂质纳米粒预处理的HUVEC细胞ICAM-1 mRNA表达量与血清中的ICAM-1蛋白含量比较较模型组有明显下降(P<0.01).结论 姜黄素脂质纳米粒剂型制备成功,结果显示该剂型可以上调内皮细胞eNOS的表达、促进NO的分泌、抑制TNF-α引起的子ICAM-1表达上升,从而发挥内皮细胞保护作用以及抗动脉粥样硬化作用.%Objective To investigate the effect of curcumin lipid nanoparticles on the expression of NO and ICAM-1 in human umbilical vein endothelial cells(HUVEC),and to explore the inhibitory effect of new dosage forms on atherosclerotic disease. Methods After curcumin lipid nanoparticles were prepared,cultured HUVEC cells were randomly divided into normal group,model group,lipid nanoparticles without drug group,three groups of dose gradient curcumin lipid nanoparticle drug group .First,10μg/L TNF-α stimulation,adding(530)mg/L curcumin lipid nanoparticles,the effect of curcumin lipid nanoparticles on the proliferation of HUVEC cells was detected by MTT assay. The gene expression levels of eN0S and ICAM-1 were determined by real-time fluorescence quantitative PCR. The expression of eN0S and ICAM-1 was observed by ELISA. Nitric acid reductase method was used to detect NO expression. Results MTT assay showed that 5-30mg/L curcumin lipid nanoparticles had no significant effect on cell viability. When the drug concentration was 60mg/L,the number of viable cells decreased.Compared with the normal group,the expression of eN0S was decreased in the model group(P<0.01). The expression of eN0S gene was significantly up-regulated by the curcumin lipid nanoparticles (P<0.01),and the secretion of NO was increased<0.01). The expression of ICAM-1 in ICAM-1 cells was increased by TNF-α. The expression of ICA-M-1 mRNA and the content of ICAM-1 protein in HUVEC cells pretreated with curcumin lipid nanoparticles Compared with the model group decreased significantly. Conclusion Curcumin lipid nanoparticles can up-regulate the expression of eNOS in endothelial cells,promote the secretion of NO,inhibit the expression of ICAM-1 in TNF-α,and play the role of endothelial cell protection and anti-atherosclerosis.
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