Effect Of Alprostadil On The Expression Of IL-6 In Urinary Exosomes Of Patients With Diabetic Nephropathy

Purpose:This study explores the expression of IL-6 in urinary exosome of DN patients,the changes of urinary protein level and IL-6 expression level in urinary exosome of DN patients after prostaglandin therapy,providing clinical ideas for further exploring molecular mechanism of DN progression and new therapeutic targets.Methods:The clinical and laboratory indexes of patients with diabetic nephropathy,who were from the two divided groups for different standards from July to November 2019 in the second hospital of Jilin University were collected,Among them,15 patients combined with alprostadil(10ug/d,1 week)under the premise of basic medication were used as the experimental group,15 patients with basic medication were used as the control group,and 200 ml of fresh urine in the morning was collected before and after one week of medication.We used the way of differential ultracentrifugation to isolate and extract urinary exosomes.Transmission electron microscope(TEM)and Western blot(WB)were the methods of identifying the exosome in our study.The concentration of the exosome was measured by BCA method,and the expression level of IL-6 on exosomes was detected by Western blot method.We use the kit method to require the m RNA on exosomes,and apply the real-time quantitative PCR(RT-q PCR)to detect the relative expression of the target gene IL-6.To clarify the effect of alprostadil treatment on clinical laboratory indexes and IL-6 expression in urinary exosomes in patients with DN.Results:1.30 patients collected from general clinical data were divided into control group andexperimental group according to whether alprostadil was used or not,15 patients in each group.The age,sex,hypertension history and smoking history of the patients were statistically analyzed between the two groups,with no statistical significance(p>0.05).2.Laboratory examination before medication,the observation points between the two groups like fasting blood glucose,urea nitrogen,blood creatinine,24-hour urine protein quantification,glycosylated hemoglobin,urine albumin/creatinine(UACR),urine microalbumin and hemoglobin have no marked difference(p>0.05).After medication,the urine microalbumin,24 h urine protein quantity and urine microalbumin/creatinine ratio in the experimental group were significantly lower than those before medication,and the difference is marked in statistical analysis when compared with the control group(p<0.05).3.We abserved the urinary exosomes obtained by the method of differential ultracentrifugation under the TEM,which were round-like and goblet-like bodies with a diameter of about 40-100 nm,The specific marker proteins CD9 and TSG101 were positive identified by WB.4.Before medication,the expression of IL-6 protein in urinary exosomes between the two groups had no statistical significance(p>0.05),the expression level of IL-6protein after intervention in the control group still had no marked change(p>0.05).While the protein expression of IL-6 in alprostadil treatment group was significantly lower than before and that in the control group(p< 0.05).5.Before medication,the m RNA levels of IL-6 in urinary exosomes of the patients in the two groups had no marked change(p>0.05).After medication,the control group in which the m RNA expression level of IL-6 in the exosomes still had no significant change(p>0.05).While the m RNA expression level of IL-6 in alprostadil treatment group was significantly lower than before and that in the control group(p<0.05).Conclusion:1.It is feasible to extract and separate urinary exosomes by differential ultracentrifugation.2.Alprostadil can reduce proteinuria in patients with early DN.3.m RNA and protein expression of IL-6 can be detected in urinary exosome of DN patients.4.Alprostadil therapy can down-regulate the expression level of IL-6 m RNA and protein in urinary exosome of DN patients.