Inflammatory Factor Induces No-reflow Through Activation Of Cyclooxygenase Signaling Pathway

ObjectiveAiming to make preliminary observation of the relationship between inflammatory factors, including C-reactive protein(CRP)or interleukin(IL)- 6, and emergency coronary blood flow before and after PCI in Acute Myocardial Infarction(AMI). By stimulating CRP concentration gradients, human coronary artery endothelial cells(HCAEC) were cultivated. Further study of effects and molecular mechanisms of inflammatory factor in cyclooxygenase(COX1, 2) signaling pathways in inflammatory factor inducing no reflow phenomenon was made, seeking a clinical predictor of no reflow, thus to further increase PCI postoperative long-dated curative effect and provide feasible and guiding treatment.MethodsClinical research: From January 2012 to January 2014, 44 acute myocardial infarction(AMI) patients were enrolled, 35 men and 9 women, aging from 31 to 91 years old, average age 60.93 ±14.46 years old.All patients’ infarction time were less than 12 hours. The diagnosis of AMI were according to the standards of the American college of cardiology(ACC) and the European society of heart disease(ESC).Inclusion criteria were:(1)a long period of time of chest pain(> 30 minutes);(2) on the standard electrocardiogram(ECG) in two or more adjacent lead st-elevation >0.20 mV;(3) the onset of chest pain 6 hours to perform PCI or remedial PCI. In order to avoid other variables affect the level of CRP, preoperative exclusion criteria were:a history of surgery or trauma less than 2 months prior to PCI, renal insufficiency(creatinine > 1.50 mg/dl), malignant tumor, febrile illness, FUO(temperature is37.5 ℃ or higher), acute or chronic inflammatory diseases and the recent infection patients. All of the patients included in the study were in accordance with complete or partial acute coronary artery occlusion, with reference to GIBSON coronary blood flow classification methods. TIMI myocardial perfusion grade(TMPG) of related artery before PCI was 0-1. preoperative venous blood were drawn. Patients were divided into two groups according to postoperative coronary artery blood flow, TMPG0-1 for no reflow group(n = 13); TMPG 2-3 level for reflow group(n = 31). All of the patients are voluntary to participate in this study, signed informed consent. Two groups of patients with general data comparison, there was no statistically significant difference(P > 0.05). To observe the relationship between the postoperative no reflow phenomenon, ELISA kit were used to determine the emergency PCI preoperative serum levels of inflammatory factors(CRP, IL- 6).Basic experiment: human coronary artery endothelial cells(HCAEC) were cultivated by stimulating CRP concentration gradients:(1) the CRP stimulatiing test:the control group, 5 minutes, 30 minutes and 60 minutes group. Namely after starvation treatment, CRP was added to HCAEC lasting 5 minutes, 30 minutes and 60 minutes respectively, and blank control group was the amount of solvent instead;(2)the extracellular regulating protein kinase(ERK1/2), c- jun amino terminal kinase(JNK1/2) inhibition test: control group, JNK1/2 inhibitors(pd98059) pretreatment group, ERK1/2 inhibitors(sp600125) pretreatment group and PD+SP group(at the same time with pd98059 and sp600125). Real Time PCR and Western Blot were used to meaasuring the levels of COX1, 2 gene transcription and protein expression, to explore the role and mechanisms of CRP in no reflow.Statistical analysis: SPSS 20.0 statistics software were applied for data processing. measurement data were expressed by x ± s, and unitized design data were checked by t test. Count data was expressed as a percentage. Multiple sets of measurement data were compared by using analysis of variance. Under the condition of population variance P < 0.05, any two groups were compared. Difference was statistically significant(P < 0.05). Chart data was drawn in GraphPad Prism 5.0.ResultsInflammatory factor(CRP) levels of patients in no-reflow group were significantly higher than those of the patients in normal reflow group, with statistically difference(P<0.05). In CRP stimulated HCAEC models, the expressions of COX1 and COX2 were elevated in CRP-treated HCAEC group compared with the control group,with statistically difference(P<0.05). After using extracellular signal regulating kinase(ERK1/2)inhibitors(pd98059) and amino terminal kinase(JNK1/2)inhibitors(sp600125), the expressions of COX1 and COX2 were decreased significantly,statistically significant difference(P < 0.01).Conclusions Triggering cyclooxygenase(COX) inflammatory pathways, C-reactive protein(CRP)can induce no reflow phenomenon after emergency PCI of the Acute Myocardial Infarction(AMI). CRP may become an important predictor of the occuring of no reflow phenomenonthe in emergency PCI of patients with AMI. Inflammatory factor ctivats the pathways of COX, and causes no-reflow in AMI patients after PCI.