Analysis Of Adipose Tissue Cytokine Profile In Chronic Kidney Disease Model With Insulin Resistance

Objective: Insulin resistance is the main component of the metabolic syndrome.It is not only common in obesity and hypertension,but also in chronic kidney disease.This study uses cytokine antibody chip technology to detect adipose tissue in mouse models of insulin resistance induced by diabetes and chronic kidney disease,analyze and compare the different cytokines of the two models,and explain the role of inflammatory factors and endothelial damage-related factors in the disease,in order to reveal the pathogenesis of insulin resistance in patients with chronic kidney disease.Methods: 1.The mouse models of chronic kidney disease and diabetic nephropathy were established respectively.The mouse model of chronic kidney disease was established by 5/6 nephrectomy.12 weeks later,according to IPGTT and ITT experiments,the model mice were divided into two groups: chronic kidney disease without insulin resistance group(CKD-NIR group)and chronic kidney disease with insulin resistance group(CKDIR group).Various biochemical indicators and pathology of kidney disease were measured to build a chronic kidney disease model.At the same time,db/db mice and db/c mice were fed with normal diet.Seven months later,urine and blood were collected to facilitate the detection of biochemical indexes,and the kidneys of each group were reserved for periodate-Schiff reaction.The pathological changes of animals were observed after staining to determine whether the model was successfully established.2.The cytokine antibody chip was used to detect epididymal adipose tissue in the above chronic kidney disease and diabetic nephropathy models.The measured cytokines with Fold Change> 1.2 and P value <0.05 were defined as differential expression factors and analysis by GO enrichment,Pathway signal pathway.Explore the role of difference factors in each model,and compare the similarities and differences between the two models.Results: 1.1.Successfully established two models of chronic kidney disease with insulin resistance:(1)Established a chronic kidney disease model: Compared with the control group,the ratio of serum creatinine,blood urea nitrogen and urinary protein creatinine in CKD-NIR mice increased,and the pathological manifestations were renal tubulointerstitial fibrosis,protein tubular type and glomerular Mesangial area widening.Compared with CKD-NIR group,blood cholesterol and insulin resistance index of CKDIR mice increased.Renal pathology showed inflammatory cell infiltration in addition to renal tubulointerstitial fibrosis,protein tube type and widening of glomerular Mesangial area.(2)Diabetic nephropathy model was successfully established: compared with db/c group,the pathological manifestations of db/db mice were tubulointerstitial fibrosis,glomerular hypertrophy,proliferation of Mesangial stromal cells,thickening of basement membrane and partial fusion of foot process of podocytes,accompanied by increased serum insulin and HOMA-IR.2.Compared with CKD-NIR mice,30 differentially expressed proteins were found in adipose tissue of CKDIR mice,mainly involved in negative regulation of hormone secretion,positive regulation of T cell chemotaxis,positive regulation of leukocyte chemotaxis,leukocyte adhesion or rolling,negative regulation of smooth muscle cell migration,stimulation of hepatocyte growth factor and other biological processes,involved in cytokine interaction,JAK-STAT signal pathway,inflammatory bowel disease,American trypanosomiasis.Signal pathways such as tuberculosis.Differential factors include: P-selectin,fibroblast growth factor receptor 3(FGFR3),interleukin 5 receptor ?(IL-5R ?),interleukin 17(IL-17),transforming growth factor-?(transforming growth factor-?(TGF-?),Lymphotactin,chemokine CCL21,Interferon ?(IFN-?),interleukin-1 receptor 9(IL-1 R9)and Granzyme G,etc.According to the molecular analysis of protein interaction network,it is found that the main factors are inflammatory factors.3.Compared with db/c mice,34 differentially expressed factors were found in db/db mice.These factors were involved in biological processes such as lipopolysaccharide response,inflammation,immune response,signal transduction,hypoxia,cell migration,interaction between cytokines,HTLV-1 infection,rheumatoid arthritis,JAK-STAT signal pathway and chemokine signal pathway.The differential factors include matrix metalloproteinase-12(MMP-12),matrix metalloproteinase-2(MMP-2),interleukin 23 receptor(IL-23),SPARC and vascular endothelial growth factor B(VEGF-B),Intercellular adhesion molecules(ICAM-1),GPNMB,chemokine LIX/CXCL5,interleukin 28(IL-28)and interleukin 11(IL-11).The molecular analysis of protein interaction network showed that the acting molecules included vasoactive factors and chemokines related to endothelial cell injury.Conclusions: Both the CKD model and the DN model had insulin resistance,but the IR adipose tissue induced by the chronic kidney disease model and the IR induced by the obese diabetic nephropathy mice had different cytokine profiles.Adipose differential factors in CKDIR mice are up-regulated factors and participate in IR-related Jak-STAT and PI3K/Akt signaling pathways.It's mainly inflammatory factors in the molecular interaction of protein network.The adipose cytokine spectrum of db/db mice includes chemokines,adhesion molecules and vasoactive factors related to endothelial cell injury,suggesting that insulin resistance induced by them has different molecular pathogenesis.