姜黄素抗动脉粥样硬化的作用及机制研究.docVIP

姜黄素对泡沫细胞形成的影响及机制 刘承，李家富，冯健，钟毅（泸州医学院附属医院心内科，四川 泸州 646000） 摘要：目的 研究姜黄素对氧化型低密度脂蛋白（oxidized low density lipoprotein,ox-LDL）诱导的人单核巨噬细胞源性泡沫细胞形成的影响及其可能的机制。方法 采用体外培养的人单核细胞白血病细胞（human acute monocytic leukemia cell line,THP-1）作为研究对象，由佛波酯（Phorbol-12-myristate-13-acetate，PMA）刺激人单核细胞为巨噬细胞，再加入ox-LDL（50 μg·mL-1）诱导复制形成泡沫细胞模型。实验分为空白对照组，模型组，姜黄素组，Toll样受体（Toll-like receptor,TLR）4抗体组，姜黄素+ TLR4抗体组，TLR4同源非特异性抗体组。采用油红O染色观察细胞形态；实时荧光定量PCR和蛋白印迹法检测TLR4的表达；酶法测定细胞内总胆固醇（total cholesterol，TC）、游离胆固醇（free cholesterol，FC）、胆固醇酯（cholesterol ester,CE）和甘油三酯（triglyceride，TG）。结果 与空白对照组比较，模型组TLR4 mRNA和蛋白表达显著增加(P＜0.05)，而姜黄素组的表达明显降低(P＜0.05)，ox-LDL刺激巨噬细胞胞内红染颗粒显著增多，且细胞大量聚集；与模型组比较：姜黄素组，TLR4抗体组，姜黄素+ TLR4抗体组胞内红染颗粒显著减少，细胞的聚集减轻，明显降低胞内TC、FC、CE和TG的含量(P＜0.05)；而TLR4同源非特异性抗体组却没有显著差别(P＞0.05)。结论 姜黄素通过抑制TLR4表达，降低胞内脂质的含量，减少人单核巨噬细胞源性泡沫细胞的形成。 关键字：姜黄素Toll样受体4氧化型低密度脂蛋白泡沫细胞 Effect and Related Mechanism of Curcumin on the Formation of Foam Cells Cheng Liu, Jiafu Li,Jian Feng, Yi Zhong(Department of Cardiology, Affiliated Hospital of Luzhou Medical College, Luzhou,Sichuan 646000,China) ABSTRACT: Objective To explore the effect of Curcumin on the formation of foam cells induced by the Oxidized low density lipoprotein(ox-LDL) and analyze the related possible mechanisms. Method As the research object, THP-1 was incubated with PMA(160 nmol·L-1) to differentiate into macrophages, then the macrophages were added with ox-LDL(50 ug·mL-1) to induce the formation of replication foam cell model. Six experimental groups were included, they were blank control group, model group, curcumin group, TLR4 antibody group, curcumin +TLR4 antibody group and TLR4 isotype control antibody group. The cell morphology was examined by Oil red staining. The expression of TLR4 was assessed buy Western Blot and Real-Time qPCR. Intracellular contents of TC, FC, CE and TG were detected through enzymatic assays. Results Compared with the blank control, the expression of TLR4 significantly increased with ox-LD added(P＜0.05), but the expression decreased with curcumi