| The incidence of type 2 diabetes is increasing worldwide.The specific predictor of type 2 diabetes has not yet been found.We construct gene chip to find the difference between normal people,impaired glucose tolerance and diabetes patients in the expression of lncRNA.Methods1 ? There are 2 healthy people,2 patients with impaired glucose tolerance and 2patients with type 2 diabetes mellitus selected,and gene chips were extracted from the whole blood to screen the differentially expressed lncRNA and mRNA and selection criteria are determined by fold change.2?GO,KEGG and other bioinformatics analysis are performed using chip data to predict possible proteins and pathways about the pathogenesis of diabetes.3? healthy people,impaired glucose tolerance patients and patients with type 2diabetes mellitus were selected and their fasting peripheral venous blood in the morning was extracted.The expression of lncRNA MEG3 in three groups was detected by real-time fluorescence quantitative PCR.4 ? At the same time,the difference of expression level of lncRNA MEG3 was detected in the islet beta cells,normal human liver cells and adipocytes in different glucose concentrations(5.5,25mmol/L).Results:1?Gene chip results showed that there were 376 differentially expressed lncRNA between the impaired glucose regulation and the normal people,and there were 448 differentially expressed lncRNA between the diabetic and the normal people and there were differentially expressed 180 lncRNA between the diabetic patients and the impaired glucose regulation patients.2?Compared with the NC group,the MEG3 in group IGR was 2 times up,3.05 times in group DM.Compared to group IGR,MEG3 in DM group was 2.2 times up;we found that MEG3 may participate in insulin resistance by consulting relevant literature,4 lncRNA(MEG3 TCL6 PVT1 GAS5)are selected for further verification.3?According to the results of the chip,199 differentially expressed lncRNA wereselected for bioinformatics analysis.In the biological process analysis,7.7% of the genes were enriched in the immune response(P<0.01).26.3% of the genes in cellular component analysis were enriched in the cell membrane(P < 0.001).According to the fold change,the first position was the methylglyoxal degradation IV pathway..4?According to the results of the chip and related bioinformatics analysis,4 lncRNA(MEG3,TCL6,PVT-1,GAS5)were selected for further verification.The PCR results showed that the expression of MEG3 in the normal human group,the impaired glucose tolerance group and the diabetic group increased in turn,and was statistically significant.The expression of TCL6 in the normal group and the diabetic group increased in turn,and was statistically significant(P < 0.05).The expression of PVT1 in DM group and impaired glucose regulation group was higher than that in normal group(P<0.05).The expression of impaired glucose regulation in LncRNA GAS5 was higher than that in normal group(P<0.05).5?The expression of MEG3 in high glucose intervention(25mmol/L)group in islet beta cells(24h)decreased by 33%(P<0.05),compared with low glucose intervention(5.5mmol/L).Under the intervention of high glucose in normal liver cells,the expression level of MEG3 decreased by 85% compared with the low glucose intervention.(P<0.05).Conclusions1?The results of gene chip showed that there were differences in lncRNA between different glycometabolism states2 ? According to the results of bioinformatics analysis of differentially expressed lncRNA,we found that these differentially expressed genes may play a role through ClpB,electron transfer,methylglyoxal degradation IV pathway and other molecules or pathways in the pathogenesis of diabetes.3 ? The expression of LncRNA MEG3 in different glucose metabolism state is different.ROC curve proves that lncRNA MEG3 has high diagnostic value as a diagnostic marker in the pathogenesis of diabetes. |
PDF Full Text Request