论文部分内容阅读
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 observe the effects of Phlegm and Blood Stasis Tongji Recipe on activation of nuclear factor-κB (NF-κB) and oxidized low density lipoprotein (ox-LDL) -enhanced human umbilical vein endothelial cells (HUVECs) (ICAM-1) expression, to explore phlegm and blood stasis with anti-atherosclerosis molecular mechanism. Methods: SD rats were randomly divided into normal group, low, medium and high dose groups of phlegm and blood stasis (24,48,72 g.kg-1.d-1) and simvastatin group (18 mg.kg-1 .d-1), prepared serum containing. HUVECs were cultured in vitro. The experiment was divided into six groups: ① normal group; ② model group; ③ low dose group of phlegm and blood stasis; middle dose group of phlegm and blood stasis; ⑤ high dose group of phlegm and blood stasis; ⑥ simvastatin group . ①, ② group with 20% normal rat serum, ③ ~ ⑥ group with 20% of each group of drug-containing serum, except the normal group to the other groups by adding 100 mg.L-1ox-LDL stimulation 3 h or 24 h after Item index determination. Real-time PCR was used to detect the expression of NF-κB p65 and ICAM-1 mRNA in HUVECs. The expression of ICAM-1 protein was detected by Western blotting. The nuclear translocation of NF-κB p65 was detected by immunofluorescence. Results: The expression of NF-κB p65 and ICAM-1 in HUVECs after ox-LDL stimulation were significantly higher than those in normal group (P <0.01). Phlegm-blood stasis and simvastatin-containing serum could significantly decrease the expression of NF-κB p65 mRNA and inhibit its nuclear translocation (P <0.05), and decrease the expression of ICAM-1 mRNA and protein (P <0.05) Statins and phlegm and blood stasis with large doses of drug-containing serum is particularly significant (P <0.01). Conclusion: Phlegm and blood stasis can inhibit NF-κB pathway in vascular endothelial cells and decrease the expression of ICAM-1, thereby reducing the inflammatory reaction, which may be one of the molecular mechanisms of anti-atherosclerosis.