| Objective In this study,lnc RNAs gene microarray technology was used to analyze the differences in peripheral blood expression of long-chain non-coding RNA(lnc RNA)and m RNA levels between patients with gestational diabetes mellitus and the control group of normal pregnant women.Bioinformatics technology was used to screen the m RNA and lnc RNAs closely related to the insulin signaling pathway in gestational diabetes mellitus,construct the lncrna-mrna network spectrum,and investigate the possible role of GDM related lnc RNAs in the development of gestational diabetes mellitus.Methods1.3 peripheral blood samples from the newly diagnosed gestational diabetes group and the control group of normal pregnant women were selected,and the basic data were recorded,respectively,to separate and extract total RNA.2.Illumina Hiseq platform and lnc RNAs gene chip technology were used to sequence the gestational diabetes group(n=3)Vs control group(n=3).The differentially expressed genes were screened with the difference multiple(FC value)greater than 1.2 times and less than0.83333 times,and P value less than 0.05 as the standard.3.Bioinformatics analysis was used to screen the m RNA and lnc RNAs that are closely related to the insulin signaling pathway of gestational diabetes mellitus and have significant differential expression,and the co-expression relationship network of lnc-RNA-m RNA was constructed.Several m RNA and lncrnas related to the GDM insulin signaling pathway were selected from them for Q-PCR verification in the gestational diabetes group(n=3)Vs control group(n=3),and bioinformatics analysis was performed on the corresponding lncrnas.4.The candidate lnc RNARPL13P5 was selected from the gestational diabetes group(n=25)Vs the normal pregnant women group(n=17)for Q-PCR amplification verification.5.The pre-pregnancy bmi,glucose tolerance test and fasting insulin of the two groups were collected,and the homeostasis model insulin resistance index(HOMA-IR)and homeostasis model insulin secretion index(HOMA-β%)were calculated to understand the level of insulin resistance and the function of islet insulin cells.6.Multiple linear regression was used to analyze the correlation between lnc RNARPL13P5 and insulin resistance in GDM.results1.Lnc RNA microarray analysis revealed that a total of 7498 genes were differentially expressed between the gestational diabetes group and the control group,including 3592up-regulated genes and 3906 down-regulated genes,and 1098 differentially expressed lnc RNAs,including 609 up-regulated genes and 489 down-regulated genes.2.The differentially expressed genes in patients with gestational diabetes mellitus peripheral blood bioinformatics analysis: GO function,according to GDM lnc RNA is mainly involved in the body’s cells carbohydrate metabolism,amino acid metabolism,regulating GTPase activity,regulating the insulin-like growth factor receptor signals,regulation of insulin receptor,pancreatic beta cells proliferation,T cell apoptosis process,etc.KEGG analysis showed that m RNA transcripts of GDM related lnc RNA were mainly enriched in PI3K/AKT signaling pathway,insulin signaling pathway,P53 signaling pathway,m TOR signaling pathway,metabolic pathway,tryptophan metabolic pathway,etc.3.According to the regulation pathway of lncrna-mrna network,4 mrnas and 6 lncrnas with significant differentially expressed key genes related to the GDM insulin signaling pathway were selected,namely IGFBP4,TSC1,TSC2,TPH1 and lnc RNAERMP1,lnc RNATSPAN32,lnc RNAMRPL38,lnc RNAAC215522.2,lnc RNARPL13P5,lnc RNASLC20A2.4.Q-PCR results of 10 candidate genes related to GDM insulin signaling pathway in the gestational diabetes group(n=3)Vscontrol group(n=3): the expressions of lnc RNA TSPAN32 and lnc RNA MRPL38 in the 6 candidate lncrnas in the gestational diabetes group were significantly higher than those in the control group(P= 0.0096,0.0371),with a P value less than 0.05,which was statistically significant.Others: the expressions of lnc RNA ERMP1,lnc RNA AC215522.2,lnc RNA RPL13P5 and lnc RNA SLC20A2 were higher than those ofthe control group(P= 0.0516,0.0840,0.0729 and 0.0538).The expression of TSC1 and TSC2 was significantly higher in the gestational diabetes group than in the control group(P= 0.0014,0.0086),and the P value was less than 0.05,which was statistically significant.Others: there was no significant difference in the expression of TPH1 and IGFBP4 in the control group(P=0.523,0.289).5.The Q-PCR results of candidate lnc RNARPL13P5 gestational diabetes group(n = 25)vs control group(n = 17)showed that the expression of lnc RNARPL13P5 in the gestational diabetes of was significantly different from that in the control group of normal pregnant women.The expression of lnc RNARPL13P5 in the peripheral blood of gestational diabetes was significantly higher,P value was less than 0.05,which was statistically significant.Multiple linear regression analysis showed that lnc RNARPL13P5,fasting insulin,insulin resistance index and islet β cell function were all involved in GDM insulin resistance.Bioinformatics analysis showed that the co expression of TSC2 in lnc RNARPL13P5 was mainly concentrated in insulin signaling pathway and PI3K-Akt signaling pathway,suggesting that lnc RNARPL13P5 played an important role in the process of GDM insulin resistance.Conclusions1.The differential expression of lncrna in the peripheral blood of patients with gestational diabetes mellitus is mainly involved in carbohydrate metabolism,amino acid metabolism,GTPase activity regulation,insulin-like growth factor receptor signal regulation,insulin receptor signal regulation,pancreatic β cell proliferation,T cell apoptosis and other processes.The m RNA transcripts of GDM insulin signaling pathway related lncrna are mainly enriched in PI3K/Akt signaling pathway,insulin signaling pathway,p53 signaling pathway,m TOR signaling pathway,tryptophan metabolism and other pathways.2.Lnc RNARPL13P5 forms a co-expression network with the TSC2 gene through the PI3K-AKT signaling pathway and the insulin signaling pathway,and participates in the pathological insulin resistance process of GDM,making an important contribution to the occurrence of GDM. |
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