Effects Of Exogenous Phosphocreatine On Mouse Inflammatory Monocyte Differentiation Induced By High Glucose And Its Possible Mechanism

Objective:Diabetes is the main risk factor affecting the morbidity and mortality of patients with coronary heart disease.There are three subgroups of mouse monocytes,among which Ly-6C^hiand Ly-6C^midare inflammatory subgroups that can secrete inflammatory factors,which damagse vascular endothelium.The differentiation of inflammatory monocyte subsets is related to atherosclerosis caused by diabetes.High glucose can induce the differentiations of circulating inflammatory monocytes and accelerate the progression of atherosclerosis,which may be related to mitochondrial oxidative stress.However,the specific intervention mechanism remains unclear.In recent years,it has been found that phosphoscreatine（PCr）,as a high-energy phosphate compound,has the characteristics of anti-mitochondrial oxidative stress and protecting vascular endothelial function.Therefore,we explore the effect of exogenous PCr on inflammatory monocyte differentiation induced by high glucose and its possible mechanism.Methods:8-week-old male clean C57BL/6 mice were used to prepare primary splenocyte suspension.Human recombinant interferon-γ（IFN-γ）was added to stimulate inflammatory monocyte differentiation.Different concentrations of glucose（20,30,40m M）were added to maintain the survival of inflammatory monocytes and induce the differentiation of inflammatory monocyte subgroups.Flow cytometry was used to observe the differentiation of monocyte subsets.Mononuclear cells（MCs）were defined as CD11b⁺MNC.According to the expression of monocyte surface antigen Ly-6C,monocytes were divided into three subsets:Ly-6C^hi,Ly-6C^midand Ly-6C^low.Results:Glucose（20、30、40m M）increases the survival of Ly-6C^hi（145%,161%and184%）and Ly-6C^mid（145%,157%and 177%）monocyte subsets in a concentration-dependent manner,but without significant effect on Ly-6C^lowmonocyte subsets.Glucose promotes the differentiation of monocyte subsets of Ly-6C^hi（144%,184%and 226%）and Ly-6C^mid（131%,167%and 199%）in a concentration-dependent manner（20,30,40m M）,but has no significant effect on Ly-6C^lowmonocyte subsets.20m M and 40m M phosphocreatine sodium significantly reduce the differentiation of Ly-6C^hi（51%and 74%）and Ly-6C^mid（43%and 71%）monocyte subsets induced by high glucose,and promotes the differentiation of Ly-6C^lowmonocyte subsets（22%and 35%）.In addition,40m M phosphocreatine sodium shows to increase the differentiation of Ly-6C^lowmonocyte subsets by 135%without glucose.Conclusion:Exogenous phosphocreatine supplement can inhibit the differentiation of inflammatory monocyte subset induced by high glucose,which may be related to the improved energy metabolism of mitochondrial.