| [Objectives]To explore the main pathways,bio-process and co-expressed IncRNA-mRNA involved in the pathogenesis of renal lesion in diabetes mice.To explore the delay effect of Yi-Qi-Bu-Shen recipe(YB)on renal lesion in diabetes mellitus(DM)mice based on lncRNA-mRNA co-expression pattern.[Methods]1.Establishment of a DM model of mice and grouping of experimental animals.Diabetes mice were induced by intraperitoneal injection of streptozotocin(STZ).Diabetic mice in good condition were randomly divided into three groups:diabetic group(DM group),low-dose treatment group of Yiqi Bushen recipe(DM +low-dose YB group)and high-dose treatment group of Yiqi Bushen recipe(DM +high-dose YB group).In addition,normal mice in good condition were selected as normal control group.2.To measure body weight(BW)and fasting blood sugar(FBS)of every mouse.To sacrifice animals and collect kidneys after 8 weeks' YB administration.3.To make histological sections and observe the histomorphological differences in different groups of mice by HE staining.4.To detect mRNAs and lncRNA expressed differentially in kidney tissue of DM group and normal control group by using chip technology.5.The mRNAs and IncRNA expressed differentially were used to conduct conjoint analysis,the GO analysis and Pathway analysis.6.Based on the results of conjoint analysis and the current research progress,a group of IncRNA and mRNA co-expressed from microarray data were selected and verified by qRT-PCR,and their co-expression in low-dose and high-dose groups was detected.[Results]1.The diabetes-induced renal lesion mice model was established successfully.Compared with the normal control group,the FBS in DM group mice were increased significantly(P<0.05).After 8 weeks'YB administration,HE staining results presented early diabetic nephropathy(DN)pathological characteristics in DM group mice,such as glomerular hypertrophy and proliferative mesangial etc.2.Compared with DM group,significant decrease of FBS and delay of pathological characteristics of early diabetic nephropathy were observed in "DM + low dose YB group" and "DM + high dose YB group" after 8 weeks',YB administration,but no significant improvement of body weight was observed.3.The results of microarray analysis showed that 148 differentially expressed mRNAs and 156 differentially expressed IncRNA were detected,38 mRNAs and 84 lncRNA were up-regulated significantly(P<0.05)while 110 mRNAs and 72 lncRNA were down-regulated significantly(P<0.05).4.GO analysis and Pathway analysis showed that the top 20 KEGG pathways were mainly TGF-beta signaling pathway,HIF-1 signaling pathway and type I diabetes mellitus,the top 20 biological processes were mainly regulation of extracellular matrix constituent secretion and apoptotic process,the top 20 cell components were mainly cytolytic granule and fibrinogen complex,etc and the top 20 molecular functions were mainly involved with oxidoreductase activity and NADP + activity etc.5.A pair of co-expressed IncRNA and mRNA were IncRNA NONMMUT066973 and SOCS2;compared with the normal control group,the expression of SOCS2 in kidney was significantly decreased while the expression of IncRNA NONMMUT066973 was significantly increased(P<0.05).The Pearson correlation analysis between the expression of SOCS2 and the expression of IncRNA NONMMUT066973 showed that the multiple correlation coefficient was-0.95(P<0.05).6.The results of qRT-PCR were similar to those of microarray.Compared with DM group,the expression of SOCS2 in kidney was significantly increased while the expression of IncRNA NONMMUT066973 was significantly decreased in "DM+low-dose YB group" and "DM+ high-dose YB group"(P<0.05).[Conclusions]1.Apoptosis,inflammation and fibrosis are the main biological pathways involved in the pathogenesis of renal lesion in DM mice;TGF-beta signaling pathway and HIF-1 signaling pathway are the main pathways.2.The expression of SOCS2 in kidney tissue of DM mice was significantly decreased while the co-expressed lncRNA NONMMUT066973 was significantly increased.Futhennore,the expression of SOCS2 was negatively correlated with the expression of IncRNA NONMMUT066973.One of the mechanisms of renal lesion in DM mice might be that the expression of SOCS2 in kidney tissue was inhibited by increasing the expression of IncRNA NONMMUT066973.3.Yi-Qi-Bu-Shen recipe can effectively control the blood sugar and delay the renal lesion in DM mice.One of the mechanisms may be that YB increase the expression of SOCS2 by inhibiting the expression of lncRNA NONMMUT066973. |
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