| Objective:To provide evidence for calcium oxalate stone formation and the mechanism of kidney injury mediated by this study.It also provides more basis for screening diagnosis and intervention targets with clinical translational potential.Methods:Establishing a rat model of calcium oxalate kidney stone,the formation of calcium oxalate crystals in the kidney was verified through HE staining and silver nitrate staining to confirm the success of the modeling.The first step is to extract the kidney samples of rats in the control group and the experimental group.Three left kidneys in each group were analyzed by transcriptome,and the candidate genes with significant differences were screened for RT-PCR validation.Finally,the potential functions were analyzed by gene ontology database and KEEG metabolic pathway.The second step is to select the right kidney samples of 3 cases in the experimental group and 3 cases in the control group for proteomic analysis,screen the differentially expressed proteins for WB and IHC validation.The protein interaction information network was constructed through the STRING database,and high score central nodes were retrieved.In the third step,combined with the above obtained transcriptome and proteome table results,the proteomic-transcriptome combination analysis was performed on the kidney samples of calcium oxalate kidney stones,and the associated differentially expressed genes and proteins were obtained.Finally,the function and regulation of differentially expressed genes and proteins in the proteome and transcriptome association analysis were screened and verified.Results:The transcriptome analysis of kidney samples from calcium oxalate nephrolithiasis rats revealed 191 transcripts that were differentially expressed,119 of which were up-regulated and 72 down-regulated.The difference,which was statistically significant(P<0.05),was confirmed by real-time PCR.GO enrichment revealed that differential gene(DEG)was mainly involved in cell processes and components;KEEG’s functional characterization revealed that DEG was mainly involved in complement and coagulation cascade reaction pathway,and the difference was also statistically significant(P<0.05).confirmed the difference,which was statistically significant(P<0.05).difference showed statistical significance(P<0.05).In the proteome analysis of kidney samples from calcium oxalate nephrolithiasis rats,the results showed that there were 330 proteins differentially expressed,190 of which were up-regulated and 140 were down-regulated.In the proteome-transcription group analysis of kidney samples from calcium oxalate nephrolithiasis rats,the results showed that in the proteome-transcription group co-up-regulated expression combination,there were 7 differentially expressed proteins,which were related to "infectious disease","immune effect",etc.In the common down-regulated expression combination of proteomics and transcriptional groups,there are also 7 differentially expressed proteins,which are mainly related to"aggravating renal tubular epithelial cell injury","respiratory system","bile excretion",etc.,P<0.05.STRING library analysis showed that Havcrl,Cst3,Lcn2,B2m and Alb were high grade central sites in the common up-regulated collection of proteome and transcriptome;In the common down-regulation collection of proteome and transcriptome,three high-scoring central sites were found,including Miox,Impa2 and Impa2.Conclusion:Calcium oxalate crystal has a significant effect on the expression of mRNA and protein in renal cells,which may play a key role in the renal injury caused by kidney stones.Spp1,Havcrl,Gpx2 and A1P1 may be diagnostic and intervention targets with clinical transformation potential.Analysis of transcriptome and proteome through association may reveal the primary genes and pathways that are expressed differently,potentially playing a critical part in the formation of kidney stones. |
PDF Full Text Request