A bleomycin induced murine model of scleroderma in GSDMD-/- versus wild type

The NLRP3 inflammasome and IL-1β are essential for scleroderma pathogenesis. Nevertheless, the role of pyroptosis executor gasdermin D(GSDMD), which is a downstream molecule of NLRP3 and is required for IL-1β release in some situations, has not yet been well elucidated in scleroderma. The purpose of this study was to explore the differences in the degree of skin fibrosis between GSDMD-/- and wild type mice in a bleomycin-induced scleroderma model, and the molecular pathways that may be involved.Total RNA from skin biopsies from GSDMD-/- and wild type mice after subcutaneous injection of BLM to induce skin fibrosis was examined by nextGen RNA sequencing GSDMD-/- and WT female mice were subcutaneously injected with BLM every other day for a total 28 days. 1x0.5cm biopsies were obtained from the skin lesions. RNA was extracted using TRIzol Reagent and quantified and qualified by Agilent 2100/2200 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA). cDNA libraries were prepared according to standard protocol and loaded on Novaseq instrument(Illumina).Sequencing was carried out using a 2x150 paired-end (PE) configuration.image analysis and base calling were conducted by the NovaSeq Control Software (NCS) + OLB + GAPipeline1.6 (Illumina) on the NovaSeq instrument.