| Background:End-stage renal disease?ESRD?is one of the major expenditures of national health care because of it's high mortality and morbidity.At present,many studies have found that ESRD hemodialysis patients have a common microinflammatory state which aggravated by increasing the risk of cardiovascular disease and other complications such as protein energy wasting?PEW?and endocrine disorders.Therefore,intervention in ESRD patients with microinflammatory state has important theoretical significance to improve its prognosis,reduce complications.Peripheral blood circulation contains micro RNAs,which usually secreted extracellular in the form of exosome.It is an endogenous non-coding RNA molecule whose length is in the 19-24 nucleotide sequence in general.We found that miR-574-5p and miR32-3P are rich in GU sequences,which are close to single-stranded RNAs?ss RNAs?that activate TLR7 / TLR8?Toll-like Receptors,TLRs?.TLRs activate transcription factors,activating inflammatory cytokines and chemokines expression by identifying and binding the corresponding pathogen-associated molecular patterns?PAMPs?.TLR7 and TLR8 signaling are dependent on the endosomal transmission of the nucleus and result in the secretion of inflammatory and regulatory cytokines by identifying ss RNAs in the nucleus or lysosomal compartment.In this study,we investigated the expression of miR-32-3p and miR-574-5p in peripheral blood of patients with ESRD and the expression of interleukin-6?IL-6?and necrosis factor-??TNF-??,and to explore the significance of miR-32-3p and miR-574-5p in the inflammatory state of ESRD hemodialysis patients,so as to provide a theoretical basis for further intervention of microinflammatory in ESRD patients.Methods:A total of 30 patients with ESRD hemodialysis and 10 healthy controls wereincluded.According to the definition of ESRD in CKD assessment and management of clinical practice guidelines,which was proposed by Kidney Disease Improvement Global Outcomes?KDIGO?in 2012.Patients who received maintenance hemodialysis?3times a week?were selected.While excluding the following patients: Age> 75 years old.In the first 3 months of enrollment,glucocorticoid and / or other immunosuppressive agents are used,or concurrent tumors,acute coronary syndromes,stroke,liver and blood system diseases,ESRD caused by autoimmune diseases,active tuberculosis,acute infection,gastrointestinal bleeding and Heart failure and other complications in patients.Healthy people with no medical history was selected as the normal control group of this study in the Second Affiliated Hospital of Dalian Medical University medical center.2ml of peripheral blood were collected in EDTA anticoagulant tube,and plasma was extracted after centrifugation.The micro RNA was extracted by the Mi RNeasy Serum / Plasma Kit from Qiagen,Germany.The relative expression of miR-32-3p and miR-574-5p was determined by real-time quantitative PCR?q RT-PCR?after reverse transcription.IL-6 and TNF-? were detected by chemiluminescence?antigen-antibody reaction?method in Second Affiliated Hospital of Dalian Medical University laboratory.All metering data is expressed by ±s and analyzed through SPSS version 19.0.If the measurement data obey the normal distribution,the two groups were compared between two independent samples t test.The Mann-Whitney U test was used for metrological data.The relative expression levels of miR-32-3p and miR-574-5p in the experimental group and the normal control group were calculated by the relative quantification method(2-??Ctmethod).The relationship between miR-32-3p,miR-574-5p and IL-6 and TNF-? was analyzed by Spearman correlation.P <0.05 for the difference was statistically significant.Results:1.Compared with the normal control group,the relative expression level of miR-574-5p was significantly up-regulated in the experimental group.The mean expression level of miR-574-5p was 11.50±11.70 in the experimental group and1.29±0.90 in the control group.The Mann-Whitney U test showed that the miR-574-5p The relative expression level was statistically significant.2.The level of IL-6 in the experimental group was significantly higher than that in the normal control group.The mean level of IL-6 in the experimental group was 15.35 ±13.49,the mean value of the control group was 1.46 ± 1.32,and the expression level of IL-6 was statistically significant by Mann-Whitney U test,P <0.05?P = 0.001?3.The level of TNF-? in the experimental group was significantly higher than that in the normal control group.The mean level of TNF-? in the experimental group was55.39 ± 48.62,the mean value of the control group was 9.38 ± 2.69,and the expression level of TNF-? in the two groups was statistically significant by Mann-Whitney U test,P <0.05?P=0.001?4.Compared with the normal normal control group,the relative expression level of miR-32-3p was not statistically significant in the experimental group.The mean expression level of miR-32-3p in the experimental group was 2.09 ± 1.63,the mean of the control group was 1.29 ± 1.05,and the expression level of miR-32-3p was not statistically significant by Mann-Whitney U test,P> 0.05 Learning meaning.5.Spearman correlation analysis?1?The relative expression of miR-574-5p in the experimental group was positively correlated with IL-6,P <0.01?P = 0.001?,and the correlation coefficient r = 0.79.?2?The results of Spearman correlation analysis showed that the relative expression of miR-574-5p in the experimental group was positively correlated with TNF-?,P<0.01?0.001?and the correlation coefficient r = 0.50.?3?Spearman correlation analysis showed that the relative expression of miR-32-3p in IL-6 was not correlated with P> 0.05?P = 0.35?and the correlation coefficient r =0.15.?4?Spearman correlation analysis showed that the relative expression of miR-32-3p in TNF-? was not correlated with P> 0.05?P = 0.19?and the correlation coefficient r=0.21Conclusions:The relative expression of miR-574-5p in ESRD hemodialysis patients was significantly higher than that normal controls,and it was positively correlated with inflammatory cytokines IL-6 and TNF-?.Micro RNA-mediated inflammatory responses may play an important role in the chronic inflammatory state of ESRD hemodialysis patients. |
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