Effect of yiqi huoxue recipe containing drug-serum on the Toll-Iike receptor-4 and its downstream signaling components MyD88 as well as the tumor necrosis factor receptor related factor-6 in human vein endothelial cells / 中国中西医结合杂志

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the influence of Yiqi Huoxue Recipe (YHR) containing drug-serum on the expression of Toll-like receptor-4 (TLR4) and its downstream signaling components MyD88, as well as the tumor necrosis factor receptor related factor-6 (TRAF-6) in human vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS), and to study its possible anti-atherosclerotic mechanism from the gene and protein levels.</p><p><b>METHODS</b>Twenty New Zealand male rabbits were equally divided into four groups in random the normal control group and the three YHR groups, 5 in each group. They were gastric perfused daily with normal saline and YHR in low, moderate and high concentration respectively. Blood drawn from rabbits' heart 2 h after ending perfusion on the 7th day, and the serum separated (that is the drug-serum) was taken for testing. HUVECs were cultured in vitro and equally divided into six groups in random the normal control group, the model group, the Western medicine group and the three YHR groups. HUVECs were stimulated with LPS, then treated separately with the drug-serum containing different concentrations of YHR for 24 h. Then the mRNA expressions of TLR4, MyD88 and TRAF-6 were measured with Real-time PCR, and their protein expressions were analyzed using Western blotting.</p><p><b>RESULTS</b>Protein and mRNA expressions of TLR4, MyD88 and TRAF-6 increased significantly after LPS stimulation (P < 0.01), but the changes in the drug-serum treated groups were significantly lower than those in the saline control group respectively (P < 0.01 or P < 0.05).</p><p><b>CONCLUSION</b>YHR can block the high expression of TLR4, and also influence the MyD88-dependent signaling pathway of TLR4, suppress the downstream expression of NF-kappaB and various related gene expressions, which may be one of its mechanisms of action for anti-atherosclerosis.</p>