| Objective: T2 DM has become one of the culprits of global health.At present,there is no specific marker for early prediction of T2 DM,so it is necessary to find biomarkers in early stage.The aim of this study was to screen for differences in the expression of micro RNAs in different states,to find potential biomarkers for T2 DM and to explore their possible role in T2 DM.Thus providing new targets for early prediction of T2 DM.Methods: 1.Two cases of healthy controls(NG),impaired glucose regulation(IGR)and type 2 diabetes mellitus(T2DM)were selected,respectively.And mi RNA gene microarray was used to screen the differentially expressed mi RNA.There was a significant difference between the two groups in the screening criteria(fold change >2 or fold change <0.5).Then mi RNAs with high expressions and high variance as well as with significant differences were selected to further confirm and study.2.108 healthy subjects,92 patients with impaired glucose regulation,117 patients with type 2 diabetes mellitus were selected,then extract morning fasting peripheral venous blood and extract serum,mi R-1005 p,mi R-574-5p,mi R-3135 b,mi R-1972,mi R-133 b and mi R-1273 f were detected by real-time quantitative PCR,so as to evaluate the predictive value of these seven mi RNAs and analyze their possible role in T2 DM by target gene prediction software and KEGG pathway analysis software.And the ROC curve was established to evaluate the sensitivity and specificity of the seven mi RNAs.Results: 1.The results of micro RNA microarray showed that there were 2549 mi RNAs between the three groups.Take fold change >2 or fold change <0.5 as the standard,there were five differentially expressed mi RNAs in the healthy control group and the impaired glucose regulation group(5 upregulated),four differentially expressed mi RNAs in the impaired glucose regulation group and the type 2 diabetes mellitus group(2 upregulated and 2 downregulated)and 24 differentially expressed mi RNAs in the healthy control group and type 2 diabetes mellitus group(17 upregulated and 9 downregulated).2.According to these differences in mi RNA between groups,the expression and the associated biological analysis,we selected 7 mi RNA(mi R-3200-5p,mi R-100-5p,mi R-574-5p,mi R-3135 b,mi R-1972,mi R-133 b,mi R-1273f)for further verification and research.The results of q RT-PCR assay showed that mi R-3200-5p,mi R-574-5p,mi R-3135 b,mi R-100-5p and mi R-1972 were decreased in healthy control group,impaired glucose regulation group and type 2 diabetes mellitus group.Among them,mi R-3200-5p and mi R-574-5p had significant difference(p<0.05)between the three groups.There was no significant difference in mi R-100-5p between healthy control group and impaired glucose tolerance group(p>0.05),and there was no significant difference in mi R-1972 between patients with impaired glucose regulation and type 2 diabetes mellitus group(p>0.05).Mi R-133 b,mi R-1273 f in the healthy control group,impaired glucose regulation group and type 2 diabetes mellitus group decreased first and then increased.However,there was a significant difference in mi R-133 b between the three groups(p<0.05),and there was no significance in mi R-1273 f between impaired glucose regulation group and type 2 diabetes mellitus group(p>0.05).Among them,the q RT-PCR results of mi R-100-5p were not consistent with the result of microarray,and the results of q RT-PCR test of the other six mi RNAs were consistent with the results of microarray.3.When mi R-574-5p,mi R-133 b,mi RNA-3135 b,mi R-1273 f and mi R-1972 were combined,the diagnosic accuracy of the control group and the impaired glucose regulation group was the highest.When mi R-3200-5p,mi R-574-5p,mi R-133 b and mi R-100-5p were combined,the diagnosic accuracy of the impaired glucose regulation group and the type 2 diabetes mellitus group was the highest.When mi R-3200-5p,mi R-574-5p,mi R-1273 f and mi R-100-5p were combined,the control group and the type 2 diabetes mellitus group was the highest.Multivariate correlation analysis showed that seven mi RNAs were associated with fasting blood glucose,glycosylated hemoglobin,body mass index,waist-to-hip ratio and systolic pressure(p <0.05).By using Tatgetscan software and KEGG pathway analysis,we can find that seven mi RNAs corresponding to their target genes played a role in protein digestion and absorption,TGF-β Signal pathway,islet secretion,p53 signaling pathway,phosphorylation signal system and other molecular biological effects and metabolic pathways.The correlation analysis showed that FBG,TG,HBA1 C,BMI,WHR were significantly increased(p <0.05),HDL-C was significantly decreased(p <0.05)in the glucose-tolerant group and type 2 diabetes mellitus group compared with the healthy control group(p <0.05),and the expression of FBG,SBP,DBP,TG,HBA1 C,WHR and BMI in type 2 diabetes mellitus group was significantly lower than that in control group(p <0.05).Conclusion: 1.The results of mi RNA microarray showed that when fold change> 2 or fold change <0.5,there were different expressed mi RNA between the three groups.2.Seven mi RNAs were screened for q RT-PCR,and seven mi RNAs were found to have significant changes in the course of diabetes mellitus,which is expected to be an early biomarker of type 2 diabetes mellitus.3.Seven mi RNAs have different diagnostic values in different combinations,and their target genes may play a role in molecular biology and metabolic pathways through protein digestion and uptake,TGF-β signaling pathway,islet secretion,p53 signaling pathway,phosphorylation signaling system and so on. |
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