The Effects Of Andrographolide On The Expression Of Chemokines And CD40-CD40L In Endothelial Cell

Abstract Objective To test the effects of andrographolide with differentconcentrations on the expression of Monocyte chemotactic protein-1、 macrophageinflammatory protein-1alpha and CD40-CD40L in human umbilical vein endothelial cellinjured by oxidized low density lipoprotein and to investigate the mechanism of antiatherosclerosis by andrographolide. Method：Neonatal umbilical cord were provided byoperation room of TongJi hospital attached to Tongji Medical College, Huazhong Universityof Science and Technology, WuHan, China. We got the human umbilical vein endothelialcell by digesting human umbilical vein with trypsogen，and cultured cells in vivo.Endothelial cells were identified by morphology and immunofluorescent response to factorⅧ related antigen. The third passage HUVECS with concentration of1×10~6/ml werecultured in ECM at37℃in a95%air and5%CO2atmosphere for24h, the cells werewashed three times with phosphate buffered saline (PBS) and maintained in M199withoutFCS for24h. The cells subsequently were divided to undergo one of the six treatments: noreagents (control),0.1mg/ml ox-LDL,0.1mg/ml ox-LDL+25mg/L andrographolide,0.1mg/ml ox-LDL+50mg/Landrographolide,0.1mg/mLox-LDL+100mg/Landrographolide,0.1mg/mLox-LDL+200mg/L andrographolide, all ox-LDL were added to the ECM afterendotheil cells were incubated with ECM with andrographolide for2h. The expression ofMCP-1、MIP-1α in HUVECS were detected by Enzyme-linked immunosorbent assay (ELISA)and Real-time quatitative Polymerase Chain Reaction (Real Time-PCR) and CD40-CD40Lwere detected by immunofluorescence. Results: As compared with the control group, theexpression of MCP-1、 MIP-1α and CD40-CD40L in ox-LDL group was increasedsignificantly（P<0.01); As compared with ox-LDL group, the expression of MCP-1、MIP-1αand CD40-CD40L by endothelial cells in AP with25mg/L and50mg/L AP groups are notchanged essentially; Cells in AP with100mg/L and200mg/L groups express less MCP-1、MIP-1α and CD40-CD40L than cells in ox-LDL group（P<0.01). Besides, the expression ofMCP-1、MIP-1α and CD40-CD40L in AP with200mg/L is reduced when comparing with APwith200mg/L groups. Conclusions: AP with above average concentrations can reduce theexpression of MCP-1、MIP-1α in HUVECS, and inhibiting the CD40-CD40L signal systemmay be its mechanism.