| Background: Diabetic nephropathy(DN)is a serious microvascular complication in diabetic patients and a common cause of end-stage renal disease(ESRD).The early prediction and diagnosis of DN is very important to improve the curative effect and prognosis of diabetic patients,and the early diagnosis and treatment of DN can delay the disease progression.Therefore,a biomarker is needed to more accurately reflect the renal function of diabetic patients.In recent years,a large number of studies have shown that exosome protein plays an important role in the pathological mechanism of diabetic nephropathy.It is expected to be a marker for early diagnosis of disease and a potential therapeutic target.Exosomes convey information to target cells through encapsulated genetic materials such as mRNA,miRNA and protein,and regulate the expression of genetic information at the transcriptional level to affect the biological functions of target cells.It may be superior to other routine detection methods in diagnosis,treatment and risk assessment.In particular,exosomal mirnas are protected by phospholipid bilayer structure and have better stability than free mirnas.However,there are few studies on the differences and functional characteristics of human plasma exosomal mirnas in DN.Therefore,to study the expression significance of plasma exosome-derived miRNA in diabetic nephropathy,and provide theoretical support for the treatment and detection of diabetic nephropathy.Objective: To investigate the potential value of plasma exosoon-derived miR-195 as a diagnostic biomarker of diabetic nephropathy by detecting the expression of plasma exosoon-derived miR-195 in patients with diabetic nephropathy and comparing it with age-and sex-matched healthy controls.Methods: Exosomes were isolated and identified by collecting blood samples from 43 patients with clinical type 2 diabetic nephropathy,20 patients without diabetic nephropathy and 20 healthy persons(control group),and then analyzed by transmission electron microscopy(TEM).The relative expression levels of exosome derived miR-195 in plasma of patients in different groups were detected by qRT-PCR.The expression and significance of miRNA in plasma of patients with diabetic nephropathy were evaluated by making ROC curve and calculating the area under the curve(AUC).Results: 1.The exosomes extracted under electron microscopy were round or quasi-round,with double-lipid structure and non-uniform size and distribution.The specific diameter was 30-150nm,and the peak diameter was 122.4nm and the mean value was 119.4nm.Most of the particle size is 30-150nm,and the actual proportion is98.5%,which is consistent with the particle size distribution of exosomes,and identified as exosomes.2.Compared with the healthy control group,the expression of plasma exosome derived miR-195 was up-regulated in both diabetic and diabetic nephropathy groups,with statistically significant differences(P<0.001).The level of exobody-derived miR-195 in diabetic nephropathy group was higher than that in diabetic nephropathy group,and the difference was statistically significant(P<0.05).3.Diabetic nephropathy patients were divided into two subgroups,stage 3 and Stage 4.The expression of exosome derived miR-195 in stage 3 diabetic nephropathy group was lower than that in stage 4 diabetic nephropathy group,the difference was statistically significant(P<0.05).4.Correlation analysis showed that miR-195 was positively correlated with ACR,urinary microprotein,Cr,fasting blood glucose,HA1c and cholesterol(r=0.454,0.732,0.299,0.298,0.435,0.470 and 0.276,respectively).miR-195 was negatively correlated with GFR(r=-0.293).miR-195 was not associated with age,BMI,diastolic blood pressure,systolic blood pressure,fasting insulin,fasting C-peptide,triglyceride and LDL.5.Multiple linear regression analysis showed that an independent correlation between HbA1c,urinary microprotein,ACR,GFR,Cr and miR-195 expression(P<0.05).6.ROC curve analysis showed that the area under the ROC curve AUC was 0.880 when the expression level of plasma exosoon-derived miR-195 was compared between the diabetic nephropathy group and the non-diabetic nephropathy group,0.945 in the diabetic nephropathy group and diabetic group,0.960 from in the diabetic nephropathy group and healthy control group.Conclusions: The expression of exosome derived miR-195 is significantly up-regulated in the plasma of patients with diabetic nephropathy at all stages,which may have potential clinical application value in the diagnosis and evaluation of diabetic nephropathy. |
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