Chronic Kidney Disease Promotes The Occurrence And Progression Of Arteriosclerosis Through Inflammatory Cells

Objective: As well recognized,the leading cause of death in patients with chronic kidney disfunction and end-stage renal disease(ESRD)is Cardiovascular disease(CVD),of which about 50% are coronary heart disease pathologically induced by atherosclerosis.Clinically,patients with chronic kidney disease(CKD)who undergo coronary angiography have been found to suffer significantly more severe lesion load than those who without chronic renal insufficiency.This phenomenon can not be explained sufficiently by traditional CVD risk factors.As atherosclerosis has been regarded as a chronic inflammatory process,in this study,we aimed to probe into new mechanisms by which CKD activate inflammatory cells including the abnormal expression of GM-CSF and abnormal sulfation modification.Thus provide new ideas and targets for preventing the occurrence and progression of atherosclerosis in CKD patients.Methods:This paper consists of three parts.The first part summarizes the epidemiology,pathogenesis,diagnosis and treatment of atherosclerotic cardiovascular diseases related to chronic renal insufficiency.In the second part,we explored the role of the colony stimulating factor GM-CSF in chronic renal insufficiency related atherosclerosis:1.A total of 1838 patients who underwent coronary angiography for the first time in the department of cardiovascular medicine were enrolled.The data of complete blood count,estimated glomerular filtration rate(e GFR)and coronary artery disease were collected,so as to establish the correlation between inflammatory cells,e GFR and the severity of coronary artery disease(gensini score was used).210 patients with different level of renal function were enrolled.Peripheral blood samples were collected and GM-CSF levels were detected.2.5/6 nephrectomy was applied to establish chronic kidney disfunction mouse model.Kidney samples were obtained.The main sources of GM-CSF in the kidney of chronic renal insufficiency mice were analyzed by histological methods.3.A chronic renal disfunction related atherosclerosis mouse model was established in Apo E knockout mice with 5/6 nephrectomy.Mice were treated with GM-CSF recombinant protein and GM-CSF neutralizing antibody to observe the effect of GM-CSF and the effect of GM-CSF neutralizing antibody.In the third part,we discussed the protein sulfation modification level in patients with ESRD and the potential mechanism of abnormal sulfation modification to promote the development of atherosclerosis:1.Peripheral blood mononuclear cells(PBMCs)were collected and protein sulfation level was analyzed by western blotting.And HPLC-MS/MS technology was applied to establish the level of O-sulfotyrosine,a metabolite in peripheral blood,to reflect the sulfation modification level of patients with ESRD.2.A chronic renal disfunction related atherosclerosis mouse model was established in Apo E knockout mice with 5/6 nephrectomy.The potential role of sulfation in the progression of atherosclerosis was observed.3.Differential modification level and expression profile of proteins in PBMCs of patients with ESRD and those with normal renal function were detected by mass spectrometry.In combination with bioinformatics analysis,the functions of abnormally modified proteins in patients with ESRD and the predicted sulfated sites were analyzed.Results:1.1 Correlation analysis showed that neutrophil/lymphocyte ratio and monocyte/lymphocyte ratio were significantly negatively correlated with e GFR and positively correlated with the severity of coronary artery lesions(gensini score).Blood GMCSF level in patients with e GFR <60 m L/min/1.73 m2 was significantly higher than that in patients with normal renal function.1.2 In the chronic kidney disease mouse model,GM-CSF mainly expressed in macrophages and endothelial cells in the papillary region of the remnant kidney.1.3 The application of GM-CSF recombinant protein significantly increase the number of peripheral blood mononuclear cells and their infiltration in the plaque in mouse model,thus promoted the plaque progression.These effects can be mitigated by GM-CSF neutralizing antibodies in chronic kidney disfunction Apo E knockout mice.2.1 In patients with ESRD,Peripheral blood metabolite o-sulfotyrosine significantly increased and the overall level of protein sulfation in PBMCs was higher,accompanied by the increased expression of tyrosylprotein sulfotransferase(TPST).2.2 In chronic kidney disfunction mouse model,the sulfation level of nucleated cells in peripheral blood significantly increased,and more infiltration of inflammatory cells in the plaques,which co-localized with the anti-sulfotyrosine antibody,was found compared to wild type mice.Furthermore,the sulfation levels of chemokine receptor CCR2 and CCR5 enhanced in peripheral blood mononuclear cells of patients with end-stage renal disease.2.3 47 proteins which were highly sulfated were found in peripheral blood mononuclear cells of patients with ESRD by Mass spectrometry and bioinformatics analysis.Sites for tyrosine sulfation of 21 proteins were searched by online predictive tools.Conclusions:1.GM-CSF released by remnant kidney in patients with renal disfunction promotes the proliferation and release of neutrophils and monocytes and their infiltration into lesions,thus promotes the occurrence and progress of atherosclerosis.This effect can be partially alleviated by GM-CSF neutralizing antibodies.Inhibition of GM-CSF may be a way to prevent atherosclerosis in patients with chronic renal insufficiency.2.The sulfation level increased in patients with ESRD which may induce the abnormal sulfation of chemokine receptors on inflammatory cells such as CCR2 and CRR5 and promote their migration and infiltration ability,which maybe a new mechanism for the progression of atherosclerosis in patients with ESRD.