【摘要】 目的探讨导痰汤通过对p38丝裂原活化蛋白激酶的调节而干预内皮细胞间黏附分子1（ICAM-1）mRNA的表达,以明确导痰汤治疗动脉粥样硬化的机制。方法取新生儿脐带,分离脐静脉内皮细胞（HUVEC）,传代培养的1～3代用于实验。含药血清的置备:将导痰汤按照0.9 g·kg^-1·d^-1剂量给SD大鼠灌胃后10d,经腹主动脉采血,分离血清。实验共分7组:HUVEC为空白对照组,含药血清（20%）处理HUVEC为导痰汤对照组,肿瘤坏死因子α（TNF-α）预处理HUVEC为TNF-α诱导组,先采用5%、10%、20%含药血清预处理HUVEC,再与TNF-α共培养为5%、10%、20%导痰汤组,使用p38特异性阻滞剂SB203580处理HUVEC为SB203580阻滞组。通过实时定量PCR和Western blot等方法,观察导痰汤对TNF-α刺激脐静脉内皮细胞培养内皮细胞ICAM-1 mRNA的表达和p38表达的影响。结果①TNF-α诱导组ICAM-1 mRNA的表达（1.17±0.26）和p38的表达（0.77±0.19）显著高于空白对照组和导痰汤对照组,差异有统计学意义（P<0.01）;使用导痰汤含药血清或SB203580处理后,5%导痰汤组ICAM-1 mRNA的表达为0.97±0.15;10%导痰汤组为0.85±0.17;20%导痰汤组为0.73±0.10;SB203580阻滞组为0.64±0.19;②5%导痰汤组p38的表达为0.69±0.21;10%导痰汤组为0.59±0.27;20%导痰汤组为0.47±0.11;SB203580阻滞组为0.36±0.09,表达显著下降,差异有统计学意义（P<0.05）;p38活性与ICAM-1 mRNA水平呈显著正相关（r=0.706,P<0.01）。结论导痰汤可以通过调节p38表达而抑制TNF-α刺激所致的脐静脉内皮细胞ICAM-1 mRNA的表达,故能起到治疗动脉粥样硬化的作用。更多还原

【Abstract】 Objective To investigate the mechanisms of daotan decoction （DTD） in the treatment of atherosclerosis by regulation of expression of intercellular adhesion molecule-1 （ICAM-1） mRNA through function of p38 mitogen activated protein kinase.Methods DTD was given to SD rats at a dose of 0.9 g·kg^-1·d^-1 through gavage.Blood samples were obtained via the abdominal aorta of SD rats 10 days after DTD gavage and the serum was separated.Neonatal umbilical cords were used to isolate endothelial cells （HUVEC）,and they were subcultured for 1 to 3 generations for use.There were 7 groups in this study:normal cultured HUVEC was blank control group;cultured with 20% DTD was DTD control group; induced with tumor necrosis factor-a （TNF-a） was TNF-a group;cultured with 5% DTD 10% DTD, 20% DTD and then TNF-α,were 5%,10%,20% DTD group,respectively;and blocked with SB203580 （p38-specific blocker） was blocking group.The impact of DTD on expression of ICAM-1 mRNA and p38 stimulated by TNF-α in HUVEC were observed with the real-time PCR and Western blot.Results ①The expression of ICAM-1 mRNA （1.17±0.26） and p38 （0.77±0.19） in the TNF-α group was significantly higher than the blank control and DTD control groups （P<0.01）;②After being treated with DTD serum, the expression of ICAM-1 mRNA in the 5%,10% and 20% DTD group were 0.97±0.15,0.85±0.17, and 0.73±0.10 respectively,and 0.64±0.19 in SB203580 group,which decreased gradually,and the three DTD groups and SB203580 group had significant difference compared with TNF-α group（P<0.05）. While the expression of p38 in the 5%,10% and 20% DTD group were 0.69±0.21,0.59±0.27, 0.47±0.11 respectively,and 0.36±0.09 in the SB203580 group,which also decreased gradually.And there was significant difference in 20% DTD group and SB203580 group compared with TNF-α group（P< 0.01）.There was a significant positive correlation between the level of p38 and ICAM-1 mRNA （r= 0.706,P<0.01）.Conclusion DTD inhibits TNF-stimulated expression of ICAM-1 mRNA through the regulation of p38 expression in HUVEC,which may play a therapeutic role in the treatment of arterial atherosclerosis.更多还原