The Mechanism Of C1Q/Tumor Necrosis Factor Related Protein1 (CTRP1) On Coronary Artery Lesions In Children With Kawasaki Disease

Background: Kawasaki disease(KD)is an immune vasculitis that mainly affects coronary arteries.In severe cases,it leads to coronary stenosis,coronary artery aneurysm,thrombosis,and myocardial infarction.At present,the pathogenesis of KD has not been completely identified,and the pathogen-induced immune response is a key factor in the pathogenesis of KD.C1 q tumor necrosis factor related protein 1(CTRP1)is a cytokine secreted by stromal vascular cells and is involved in inflammatory diseases and vascular diseases.The purpose of this study was to investigate the role of CTRP1 in the acute phase of KD and the coagulation process after coronary artery injury.Methods:(1)We Collected serum samples from children with acute KD and normal children,and measured serum concentrations of CTRP1 in all subjects and serum concentrations of tumor necrosis factor ?(TNF-?),interleukin 1?(IL-1?)and IL-6 in the children with acute KDby Enzyme-linked immunosorbent assay(ELISA);(2)Adiponectin receptor 1,(AdipoR1)inhibited human coronary artery endothelial cell(HCAEC)was constructed.HCAEC and AdipoR1 inhibited HCAEC were stimulated by recombinant CTRP1.The expression of AMPK,Akt and eNOS phosphorylation protein was measured by Western-blot.The expression of AMPK,Akt and eNOS mRNA were measured by Real-time.NO levels in the culture medium was measured by chemical colorimetry;(3)Single intraperitoneal injection of Lactobacillus casei wall extract(LCWE)to construct a mouse model of coronary artery inflammation and intraperitoneal injection of recombinant CTRP1 on the coronary arteritis model mice.The collagen exposure in the coronary artery was observed by Masson staining,and serum levels of fibrinogen(FIB),thrombin(TB),thromboxane B2(TXB2)and D-dimer(DD)at different time points after CTRP1 intervention were measured by ELISA.Results:(1)Compared to the Healthy group,serum CTRP1 levels were significantly elevated in the KD group.Significantly higher serum TNF-?,IL-1?,IL-6,and CTRP1 levels were observed in patients with KD with coronary artery lesions(KD-CAL)than patients with KD without CAL(KD-NCAL).Serum CTRP1 levels were positively correlated with white blood cell counts(WBC),percentage of neutrophils(N%),thrombin time(TT),procalcitonin(Pct),TNF-?,IL-1?,and IL-6levels.Meanwhile,CTRP1 levels were negatively correlated withpercentage of leukomonocytes(L%)in KD patients.Furthermore,serum CTRP1 levels were positively correlated with the timepoint of intravenous immunoglobulin(IVIG),WBC,N%,TNF-?,IL-1?,and IL-6levels in KD-CAL group.(2)Compared with Blank group,the phosphorylation proteins and mRNA expressions of AMPK,Akt and eNOS induced by CTRP1 and adiponectin were significantly increased in HCAEC,while the they were decreased with AMPK,Akt,eNOS inhibotors;NO levels secreted by HCAEC were increased than those in blank group with CTRP1 stimulation,while there was no significant difference with adiponectin and AMPK,Akt,eNOS inhibotors.AMPK,Akt,eNOS phosphorylation and mRNA were lowly expressed in the AdopoR1 inhibited HCAEC with CTRP1 stimulation,and there were no significantly difference with AMPK,Akt,eNOS inhibotors;NO levels secreted by AdopoR1 inhibited HCAEC were not increased with CTRP1 stimulation,and there were no significantly difference with adiponectin,and AMPK,Akt,eNOS inhibotors.(3)After CTRP1 injection,the collagen exposure around the coronary artery was significantly improved over time,and gradually returned to normal in 7-14 days;After CTRP1 injection,the serum FIB and TB levels in mice did not change significantly in 1-3 days,and increased significantly in 7-14 days.The serum DD of mice did not change significantly after 1 day,and decreased significantly from the third day.There was no significant change in thelevel of decline in 3-14 days.The serum TXB2 in mice showed a downward trend from 1 day to 14 days,but the level of decline was not statistically significant.Conclusion:(1)CTRP1 acts as an inflammatory factor in the inflammatory process of Kawasaki disease and mediates coronary artery lesion in the acute phase of Kawasaki disease;(2)CTRP1 binds to AdipoR1 and activates AMPK/Akt/eNOS signaling pathway to induce abnormal secretion of NO by HCAEC;(3)CTRP1 can improve collagen exposure after inflammatory injury in coronary artery,resulting in changes of serum FIB,TB and DD levels.