The Effect Of Lox-1on The Expression Of Tissue Factor Induced By C-Reactive Protein In Human Umbilical Vein Endothelial Cells

Background: At present,C-reactive protein is considered to be anindependent and strong cardiovascular disease risk factors，C-reactiveprotein induces tissue factor upregulation in vascular endothelialcells.Objective:To investigate the effect of lectin-type oxidized LDLreceptor1（LOX-1）on expression of tissue factor(TF) induced by C-reactiveprotein (CRP)in Human Umbilical Vein Endothelial Cells(HUVECs), andto explore the possible role of CRP in the arterial thrombosis.Methods: Recombinant plasmid pTracer CMV2-CRP was constructedand was transfected into HUVEC cells，then the levels of transcription andprotein of CRP in HUVECs were identified by PCR and Western blot.After the recombination plasmid pTracer CMV2-CRP transfected intoHUVECs, HUVECs were divided into a normal control group、GFP groupCRP-GFP group, RT-PCR was used to detect the mRNA of LOX-1andTF,and Western blot was used to detect the protein expression of LOX-1、 TF、P-ERK1/2、T-ERK1/2、P-JNK、and T-JNK respectively.Incubationof HUVECs transfection CRP with anti-LOX-1or inhibitors to ERK1/2（U0126）or inhibitors to JNK(SP600125),the expression of TF genes wasdetected by using RT-PCR and Western blot.Result: IN CRP plasmid transfected HUVEC cells, green fluorescentprotein was found and CRP mRNA and protein expression were detected.Transfection of human HUVECs with CRP expression plasmidsignificantly increased LOX-1mRNA and protein levels（0.70±0.03vs0.24±0.01,0.6616±0.0200vs0.2558±0.0400, P＜0.05）and TF mRNA andprotein levels(0.442±0.040vs0.146±0.030，0.7274±0.0200vs0.2174±0.0200,P＜0.05). CRP activated ERK1/2, but not JNK MAPkinase (MAPK), and the effect of CRP on TF expression was blocked bypharmacological inhibitor of ERK1/2（U0126）, but not JNK（SP600125）MAPK.neutralizing antibody to LOX-1could significantly attenuatemRNA and protein expression of TF induced by CRP（0.264±0.030vs0.442±0.040,0.3244±0.0400vs0.7274±0.0200,P＜0.05）.Conclusion: The recombinant plasmid pTracer CMV2-CRP wassuccessfully transfected into HUVEC cells, and CRP gene was successfullytranscripted and translated in the HUVECs transfected by CRPplasmid.We conclude that CRP enhances HUVECs LOX-1expression andpropose a new mechanism by which CRP may promote HUVEC TFexpression, that of inducing LOX-1，which may be related to ERK1/2 MAPK but not JNK MAPK pathway.