Abstract:

Objective Based on the GEO database，to analyze and verify the differentially expressed genes in patients with chronic kidney disease（CKD） by cytological experiments in order to find new potential targets for CKD. Methods The microarray data related to CKD（GSE66494 dataset，GSE15072 dataset and GSE30122 dataset） were downloaded from GEO database. The differentially expressed genes in GSE66494 dataset and GSE15072 dataset were screened by bioinformatics methods. Hub genes were identified by constructing protein-protein interaction（PPI） network. The expression of selected hub genes was analyzed in the GSE30122 dataset. The NetworkAnalyst database was used to construct a microRNA-hub gene regulatory network，and the top 5 miRNA with high association with hub genes were screened. The proximal telomeric renal tubular epithelial cell line HK-2 was used to construct a CKD cell model. According to the different substances added，they were classified into tumor necrosis factor-alpha（TNF-α） group（100 μg·L^-1 TNF-α），TNF-α+infliximab group [10 ng·mL^-1 infliximab（TNF-α specific inhibitor）+100 μg·L^-1 TNF-α] and control group（equal proportion of solvent dimethylsulfoxide）. The expression of hub gene and microRNA in each group was determined. Results A total of 233 differentially expressed genes were screened，which included 20 up-regulated genes and 213 down-regulated genes. Totally，10 hub genes（HSP90AA1，IGF1，REN，MMP2，ACTA2，PXN，COL3A1，CALCA，DMD，LAMA2） and 5 microRNAs（hsa-miR-29b-3p，hsa-miR-421，hsa-miR-29a-3p，hsa-miR-335-5p and hsa-let-7b-5p） were screened out. The analysis results of the GSE30122 dataset showed that the hub gene was more significantly expressed in renal tubular tissue samples. Compared with control group，TNF-α group showed obvious cell necrosis and decreased cell growth density. The relative expression levels of HSP90AA1，IGF1，COL3A1，CALCA and hsa-miR-29b-3p，hsa-miR-421，hsa-miR-29a-3p were increased（P<0.05）. Compared with TNF-α group，the cell necrosis in TNF-α+infliximab group was improved，and the cell growth density was increased. The relative expression levels of HSP90AA1，COL3A1 and hsa-miR-29b-3p，hsa-miR-421，hsa-miR-29a-3p were decreased（P<0.05）. Conclusions There are 10 hub genes and 5 microRNA selected from GEO database. HSP90AA1，IGF1，COL3A1，CALCA and hsa-miR-29a-3p，hsa-miR-29b-3p，hsa-miR-421 may be involved in the process of CKD development，which can be potential targets for CKD.

Key words: Differentially expressed gene, MicroRNA, Chronic kidney disease