| Background and purposeDiabetic microangiopathy, including kidney disease, neuropathy, retinopathy. Risk factors Complications development affected by many factors, including diabetes duration, and genetic factors.[1] One of diabetes of the most common microvascular complications of diabetic nephropathy (DN), is an important cause of ESRD in Western countries and death in patients with DM is the leading cause of ESRD. in China, DN also lead to increased year by year in the number of ESRD.[2] The current DN pathogenic mechanism is not entirely clear, effective treatment is still inadequate, therefore, effective early intervention and prevention, treatment DN is especially as the population ages, the number of population-based diabetes medical field increase in the incidence of diabetic nephropathy will also increase, which will be extremely important. Regardless of type1or type2diabetes can lead to vascular complications, in addition to high blood sugar, high LDL, advanced glucose metabolism, growth factors and factor authentication, the need for better understanding of the molecular mechanisms of diabetes to determine the new therapeutic goal.[3] The understanding underlying the pathogenesis of diabetic complications, such as chronic diseases and improving the status of development of new therapeutic strategies are still very important.New methods from nematodes identified a novel endogenous small fragments, single-chain, non-coding RNA molecules, named micro RNA, micro RNA such as a growth regulating factor.[4] miRNAs are small RNA, ubiquitous in eukaryotes. From a precursor (pre-miRNA) is further mature miRNA, particularly endogenous miRNA hairpin structure length of70-80nt precursor (pre-miRNA) is generated after the cutting, the mature miRNA is a class of about21-24nt through the negative regulation of transcription of a target gene expression of non-protein coding RNA molecules that can bind the target gene3’-untranslated region,3’-UTR and the complementary binding of the target mRNA expression regulation.[4] they are present in the fluid circulation miRNA specific extracellular existence. Found in human serum, plasma, urine, saliva, breast milk and other body fluid can be detected more microRNAs. Most microRNA molecule in a body fluid is not present in free form, it is a lipid, protein complexes, microvesicles (MV), the outer clamp member (exosome) parcels and other binding, so it has good resistance to RNase degradation, high stability.[5] After the cells are the source of biological factors to stimulate selectively wrapped into the specific microRNA MV, also while carrying a cell surface marker on the surface of MV. In different tissues, cell surface marker antibody beads can be easily detected in different tissue sources MV and microRNA. It is because of the characteristics of serum circulating microRNA, it is ideally suited for early warning and early diagnosis as a biomarker for the disease. Early changes in the change of its target gene, the stability of the body fluid, the running properties and the specific characteristics of the tissue of origin cells, early warning and it is very suitable as a biomarker for the diagnosis of disease.[24] Since the1950s, the discovery of the relationship messager RNA, protein and mRNA between the biopolymers have been major research and protein synthesis in the transfer of genetic information. And recent research in the regulation of mRNA, especially the role of miRNA in hot regulation of mRNA. MiRNA is a short RNA sequence, the hairpin structure due to the first Pre-miRNA, which play a key role in the transcriptional regulation, by forming a dual role, is fully complementary to the target mRNA of the target gene is cleared, it is not completely complementary to the target inhibition genes.[15] microRNA as a potential carrier protein delivery and stable expression of the endogenous RNA dynamic competition network. miRNAs can induce protein synthesis threshold, the threshold can yield aggregation of the protein transcription factor to complete the reaction. Therefore, regulation of miRNA small but play the role of protein synthesis.[16] DNA encoding mRNA and then translated into protein, miRNA target gene mRNA restrain or remove the effect, so miRNAs may be negatively correlated with the protein. So some may suspect that the increase may be related to inhibition of protein to reduce its miRNAs, and therefore detected in normal controls reduce the miRNA may be more meaningful. In diabetic nephropathy in renal pathological changes between extracellular matrix deposition in glomeruli and renal interstitial site is one of the factors that lead to sustained progression of diabetic kidney,[2] may inhibit the synthesis of extracellular matrix miRNA reduced. Based on the theoretical understanding of diabetic nephropathy and diabetic nephropathy miRNAs proposed mechanism is closely related to serum microRNA may occur, and the cycle microRNA decrease compared with normal healthy person may serve as early warning and biomarkers for early diagnosis of diabetic nephropathy. Under the guidance of this idea, we plan to conduct spectrum detection of serum microRNA diabetes.Subjects and Methodsby selected age and sex-matched patients with diabetic nephropathy, diabetic, healthy control group of20people by miRCURY TM LNA expression array screening (KangChen company) three groups differences microRNAs further combined database analysis miRBase, TargetScan Human, miRtarBase may be associated with the pathogenesis of diabetic nephropathy associated miRNAs15a further investigate the role of miRNAs in the pathogenesis of DN foundation by RT-PCR validation and discussion of research and follow-up. further validation by RT-PCR and use2-ΔΔCT multiple relationship between Δ CT analysis group; used SPSS17.0software one-way ANOVA ANOVA and correlation with clinical data analysis.Currently mechanism in diabetic nephropathy serum microRNA less, to carry out this study reveals microRNA mechanisms of diabetic nephropathy in patients with diabetes, and is expected to determine the1-2as a warning marker serum microRNA.Result1, miRCURY TM LNA expression array results:According microRAN array signal intensity differences between the three groups there are more than two times: Diabetic nephropathy group than in the healthy group hike microRNAs158downregulated in microRNAs150months; diabetic nephropathy group than in the diabetic group hike microRNAs14downregulated in microRNAs6months; diabetic group than the healthy group raised106, down125.2, Application of bioinformatics to obtain in miRBase, miRtarBase, TargetScan database may be related to the pathogenesis of diabetic nephropathy microRNAs15months, hsa-miR-150-5p, hsa-miR-485-3p, hsa-miR-205-3p, hsa-miR-302e, hsa-miR-495-5p, hsa-miR-361-3p, hsa-miR-377-3p, hsa-miR-484, hsa-let-7a-2-3p, hsa-miR-532-3p, hsa-miR-302a-3p, hsa-miR-519e-3p, hsa-miR-301a-5p, hsa-miR-216a-3p, hsa-miR-328-3p.3, RT-PCR validation:were designed15microRNAs primers, according to QIAGEN’s miRNAs internal reference miR-39as an internal reference, by RT-PCR validation, diabetic nephropathy miR-150was significantly less than in the healthy group, P<0.05, The diabetic group and healthy group meaningless.Conclusion:patients with diabetic nephropathy circulating miR-150compared with the healthy control group decreased in line with the miRCURY TM LNA expression array chip results. The miRNA-150May be as a biomarker for early warning of diabetic nephropathy... |
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