Objective: To observe effect of Huoxue injection on the expression of intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), the adherence of monocytes to endothelial cells, and the regulation role of nuclear factor kappa B (NF-kappaB) in cultured human umbilical vein endothelial cells (HUVEC) injury induced by the oxidized low-density lipoprotein (ox-LDL).
Method: The ox-LDL (100 mg x L(-1)) was added to the cultured HUVEC to prepare the injury model of HUVEC. The adhesive percentage between HUVEC treated with ox-LDL and monocytes was determined by protein quantification. Expression of mRNA and protein of ICAM-1 and VCAM-1 were determined by RT-PCR and flow cytometry respectively. The percentage of positive cells and the ratio of nuclei and cytoplasm of NF-kappaB p65 staining in HUVEC the were examined by cell immunochemistry.
Result: Treatment of HUVEC with ox-LDL for 12, 24 hours significantly increased adhesion of monocytes to HUVEC and enhanced the expressions of mRNA and protein of ICAM-1 and VCAM-1. The percentage of positive cells and the ratio of nuclei and cytoplasm of NF-kappaB p65 staining in HUVEC were significantly increased after treatment with ox-LDL for 24 hours. Huo Xue Injection could significantly inhibit the adhesion between monocyte and HUVEC, the expression of mRNA and protein of ICAM-1 and VCAM-1, and declined the percentage of positive cells and the ratio of nuclei and cytoplasm of NF-kappaB p65 staining in HUVEC. The effects were strengthened with increasing the deal of Huoxue injection.
Conclusion: Huoxue injection has an inhibitory effect on the adherence of monocytes to HUVEC, probably by way of down-regulating the expression of mRNA and protein of ICAM-1 and VCAM-1 in HUVEC. The mechanism is probably associated with inhibiting the activation of NF-kappaB p65 of HUVEC. The effects of Huoxue injection can bring about the protective effect to endothelial cells injury induced by ox-LDL.