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RNA-sequencing of mouse diabetic wounds treated with mast cell stabilizer MCS-01

ABSTRACT: To investigate global changes in gene expression patterns following treatment with DSCG or MCS-01 we performed RNA-sequencing and nanostring profiling analyses on Day 10 wounds. Differential expression analysis on pre-MCS-01 samples compared to blank identified 649 significantly altered genes at p value <0.05 and absolute fold change >=2. Similarly, in post-MCS-01 samples 954 genes were differentially expressed, of which 566 were down- and 388 up-regulated. DCSG treatment resulted in differential expression of 91 genes. RT-qPCR corroborated increased expression of NFκB and STAT3 with post-MCS-01 treatment. In addition, we compared miRNA expression between MCS-01 samples and controls. Pre-treatment significantly altered expression of 11 miRNAs, while post-treatment altered expression of 28 miRNAs. RT-qPCR confirmed miR-34c upregulation, while significant inverse correlations were found between miR-34c and selected target genes Lef-1, Myb, and Mycn.

ORGANISM(S): Mus musculus

PROVIDER: GSE135915 | GEO | 2019/10/25

REPOSITORIES: GEO

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Project description:Cerebral ischemia/reperfusion injury (CI/RI), including neurological behavior deficits, cerebral infarction, blood-brain barrier (BBB) dysfunction, and neuroinflammation, et al. is a severe challenge in treatment of ischemic stroke. Traditional Chinese medicine (TCM) with the characteristics of multi-components and multi-functions for multiple targets/pathways has been historically used in the treatment of stroke and post stroke recovery in China. QiShenYiQi (QSYQ), a representative component-based Chinese medicine, is capable of reducing organ injury caused by ischemia/reperfusion via multiple mechanisms. Recently, we have previously reported that the positive action of QSYQ against the acute phase of CI/RI is partly via modulation of neuroinflammatory response. However, the effects and underlying mechanisms of QSYQ on subacute phase of CI/RI remain unknown, which are aimed to investigate in present study. The pharmacological action of QSYQ treatment on brain damage was estimated in the experimental stroke rats underwent 90 min ischemia and 8 days reperfusion by assessing the neurological and locomotor deficits, cerebral infarction, brain edema, and BBB integrity. Furthermore, we used TMT-based quantitative proteomics technology to identify significantly differentially expressed proteins following QSYQ treatment, which could give important clues for revealing the possible mechanism underlying the positive role of QSYQ in CI/RI. Besides, immunohistochemistry, western blot analysis, RT-qPCR, and rat ELISA kits were executed to further verify the accuracy of proteomics analysis results. As a consequence, we found that treatment with QSYQ (600mg/kg) for 7 days obviously improved neurological and behavioral impairment, attenuated infarct volume and brain edema, and alleviated BBB breakdown in stroke rats. Bioinformatics analysis suggested that differentially expressed protein galectin-3 mediated inflammatory response was closely related to the beneficial effect of QSYQ. Results of immunohistochemistry, western blot analysis and RT-qPCR showed that the model rats treated with QSYQ (600mg/kg) markedly downregulated the mRNA and protein expression level of galectin-3 in brain tissues compared to the untreated stroke rats. In addition, the decrease of mRNA level in brain tissues and contents in serum of TNF-α and IL-6 following QSYQ treatment were also observed. These interesting finding manifested that the effective action of QSYQ against subacute phase of CI/RI was partly via regulating galectin-3 mediated inflammatory reaction.

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RNA-sequencing of mouse diabetic wounds treated with mast cell stabilizer MCS-01

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