摘要
目的:观察痰瘀同治方含药血清对氧化型低密度脂蛋白(ox-LDL)损伤的人脐静脉内皮细胞(HUVECs)核因子-κB(NF-κB)的活化和细胞间黏附分子-1(ICAM-1)表达的影响,探讨痰瘀同治方抗动脉粥样硬化的分子机制。方法:SD大鼠随机分为正常组,痰瘀同治方低、中、高剂量组(24,48,72 g.kg-1.d-1)和辛伐他汀组(18 mg.kg-1.d-1),制备含药血清。体外培养HUVECs,实验分为6组:①正常组;②模型组;③痰瘀同治方低剂量组;④痰瘀同治方中剂量组;⑤痰瘀同治方高剂量组;⑥辛伐他汀组。其中①、②组用20%正常鼠血清,③～⑥组用20%各组含药血清,除正常组外其余各组加入100 mg.L-1ox-LDL刺激3 h或24 h后进行各项指标测定。Real-time PCR法检测HUVECs NF-κB p65和ICAM-1mRNA表达,Western blotting检测ICAM-1蛋白表达,细胞免疫荧光法检测NF-κB p65核移位变化。结果:HUVECs经ox-LDL刺激后NF-κB p65和ICAM-1的表达与正常组比较均明显升高(P<0.01)。痰瘀同治方和辛伐他汀含药血清能显著降低NF-κB p65 mRNA表达及抑制其核移位(P<0.05),降低ICAM-1 mRNA和蛋白表达(P<0.05),其中以辛伐他汀和痰瘀同治方大剂量含药血清作用尤为显著(P<0.01)。结论:痰瘀同治方能够通过抑制血管内皮细胞NF-κB通路,降低ICAM-1表达,进而减少炎症反应,这可能是其抗动脉粥样硬化分子机制之一。
Objective：To explore the anti-atherosclerosis（AS） effect of serum containing Tanyu Tongzhi Fang（TYTZF） on the activation of nuclear factor-kappa B（NF-κB） and the expression of intercellular adhesion molecule-1（ICAM-1） in human umbilical vein endothelial cells（HUVECs） injured by oxidized low density lipoprotein（ox-LDL）.Method： SD rats were equally divided into five groups in random： the control group,TYTZF low dose group,TYTZF middle dose group,TYTZF high dose group（24,47,72 g·kg-1·d-1） and simvastatin group（18 mg·kg-1）.After administration,the serum was taken for testing.HUVECs were treated with serum containing TYTZF and simvastatin respectively and incubated with ox-LDL（100 mg·L-1） for an additional 3 h or 24 hours.The mRNA levels of NF-κB p65 and ICAM-1 were measured by real-time PCR and the protein expression of ICAM-1 was detected by Western blotting.The activation of NF-κB p65 was observed with immunofluorescence method.Result： The expression of NF-κB p65 and ICAM-1 in serum containing TYTZF group and simvastatin group were significantly lower than that in modle group（P0.05）,especially in the high-dose serum containing TYTZF group and simvastatin group（P0.01）.Conclusion： The mechanism of TYTZF anti-AS may be related to the inhibitory effect of NF-κB pathway,thereby reducing the expression of ICAM-1 in vascular endothelial cells.
出处
《中国实验方剂学杂志》
CAS
北大核心
2012年第5期140-144,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家"十一五"科技支撑"重大新药创制专项课题"(2009ZX09502-017
2009ZX09301-005
2009ZX 09301-005-2-6)
