[Effects of monocyte chemotactic protein-3 on ICAM-1, VCAM-1, TF, and TFPI expression and apoptosis in human umbilical vein endothelial cells].

OBJECTIVE: To investigate the effect of monocyte chemotactic protein-3 (MCP-3) on the expressions of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), tissue factor (TF, and tissue factor pathway inhibitor (TFPI) and cell apoptosis in human umbilical vein endothelial cells (HUVECs). METHODS: Cultured HUVECs were treated with MCP-3 at the optimal concentration determined previously 1 h after treatments with or without MCP-3 antibody (20 ng/ml), PI3K inhibitor, or LY-294002 (5 mmol/ml). The expressions of ICAM-1, VCAM-1, TF and TFPI were analyzed using RT-PCR and Western blot after the treatments. MCP-3 mRNA and protein expressions were detected in HUVECs exposed to 50 µg/ml ox-LDL for 24 h. The cell apoptosis and caspase-3 protein production in HUVECs treated with MCP-3 or with MCP-3 plus CCR2 antagonist for 24 h and 48 h were evaluated by flow cytometry and Western blotting. RESULTS: At the optimal concentration of 0.3 ng/ml, MCP-3 treatment for 24 h caused significantly increased ICAM-1, VCAM-1, and TF expressions with lowered expression of TFPI in HUVECs (P<0.05), and such effects were significantly inhibited by the application of MCP-3 antibody, PI3K inhibitor, or LY-294002 (P<0.05). Ox-LDL exposure significantly increased the expression of MCP-3 in HUVECs (P<0.05). HUVECs showed a significantly increased apoptosis rate after treatment with MCP-3 or with MCP-3 plus CCR2 antagonist (P<0.05), and the apoptosis rate increased significantly as the treatment time prolonged (P<0.05); caspase-3 protein expression in the cells showed a similar pattern of alterations following the treatments. CONCLUSION: ox-LDL can induce MCP-3 expression in HUVECs. MCP-3 induces apoptosis of HUVECs and significantly affects the cellular function partially through the PI3K signaling pathway.

[Effects of Bushen Huoxue Fang on rat cardiac fibroblast proliferation and collagen production in vitro].

OBJECTIVE: To investigate the effects of Bushen Huoxue Fang on the proliferation of rat cardiac fibroblasts and collagen production in the cells. METHODS: Rat cardiac fibroblasts were isolated and cultured in DMEM containing 10% (group A) or 20% (group B) or no (group C) serum from rats treated with Bushen Huoxue Fang, with cells cultured in DMEM containing 10% FBS as the control (group D). After 72 h of cell culture, the proliferation of the fibroblasts was detected using CCK-8 kit, and collagen mRNA and protein expressions were examined using RT-PCR and Western blotting, respectively. RESULTS: Compared with that in groups C and D, the cell proliferation decreased significantly in groups A and B, and especially in the latter (P<0.05). RT-PCR demonstrated significant reductions of the mRNAs of type 1 and 3 collagens in groups A and B (P<0.05), and their protein levels were also significantly lowered (P<0.05). CONCLUSION: Bushen Huoxue Fang can effectively inhibit the proliferation of rat cardiac fibroblasts and reduced collagen type 1 and 3 productions in the cells in vitro.