Transcriptomics analysis of sirolimus treatment in lupus nephritis

Abstract

lupus nephritis (ln) is one of the principal causes of mortality and disability in patients with systemic lupus erythematosus. Sirolimus has been used to treat patients with ln; however, the effects and mechanism of sirolimus in these patients remains unclear. The present study aimed to elucidate the therapeutic effects and mechanisms of sirolimus in ln mice using low, medium and high doses of sirolimus (0.1, 0.3 and 1 mg/kg, respectively). The survival probability and kidney index were calculated, and renal fibrosis was determined using Masson's Trichrome staining. The expression levels of e-cadherin, α-smooth muscle actin (α-SMa) and vimentin were assessed via immunofluorescence analysis. Transcriptome analysis of control and sirolimus-treated ln mice was performed using rna-sequencing, differentially expressed gene (deG) identification and annotation, and Gene ontology (Go) functional and Kyoto encyclopedia of Genes and Genomes (KeGG) pathway enrichment. The results suggested that a medium dose of sirolimus alleviated renal fibrosis and increased the survival rates of mice with LN (P<0.05). Furthermore, transcriptome analysis revealed 334 deGs associated with ln, 176 of which were upregulated and 158 were downregulated. Following Go functional enrichment, 'biological process', 'molecular function' and 'cellular component' terms were identified. a total of 10 KeGG pathways were enriched, with 'cytokine-cytokine receptor interaction' and 'interleukin-17 signaling pathway' being significantly enriched (P<0.05). To the best of our knowledge, the present study is the first to conduct transcriptome analysis of ln mice treated with sirolimus, and demonstrated that a dose of 0.3 mg/kg exerted the greatest therapeutic effects.

Tacrolimus increases the expression level of the chemokine receptor CXCR2 to promote renal fibrosis progression

“…In addition, it has previously been reported that the epithetlial-mesenchymal transition (EMT) is significantly correlated with renal fibrosis, which is associated with calcineurin inhibitor-mediated nephrotoxicity (21). EMT is one of the basic mechanisms of renal fibrosis and involves various processes in which epithelial cells stop exhibiting epithelial characteristics, including the expression of E-cadherin, and obtain traits specific of mesenchymal cells, including the upregulation of α-SMA (25,47-48). In addition, it has previously been reported that vimentin is a potential novel biomarker in renal fibrosis (25,49).…”

“…After Wistar rats were sacrificed, the kidneys were removed and weighed. The kidney index was calculated using the following formula: Kidney index = (average kidney weight/body weight) (25). Paraformaldehyde (4%) was used to fix the fresh kidneys, and liquid nitrogen was used to rapidly freeze the residual kidney tissues, which were stored at -80°C until further analysis.…”

“…Suzhou Basepair Biotechnology Company (http://www.basepair.cn/) performed the RNA library construction and sequencing in biological triplicates using an Illumina HiSeq X Ten Sequencing system (Illumina, Inc.). The main sequencing processes included sequencing data quality preprocessing, reference genome alignment, gene expression analysis and differential expression analysis (25,27).…”

“…To obtain clean reads, adaptor-contaminated and low-quality sequences were removed using filtering methods, as previously described (28). FastQC (version 0.11.4; http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) was used to examine the quality of the clean reads (25). Subsequently, the reads were aligned to the reference genome Ensemble-Rnor6.0 using Hisat2 (version 2.1.0).…”

“…KEGG pathway analysis was performed for the DEGs using the KEGG Orthology-Based Annotation System (31). The significantly enriched KEGG pathways exhibited a P<0.05 (25,27,28,32).…”