Project description:Purpose: The goals of this study was to (1) evaluate the protective effect of celastrol on alpha-naphthylisothiocyanate (ANIT)-induced cholestasis and (2) which genes were recovered by celastrol. Methods:To investigate the protective effect of celastrol on ANIT-induced cholestasis, the WT mice were randomly assigned into two groups, respectively (n=3): (1) ANIT; (2) ANIT+Celastrol. ANIT+Celastrol group was orally treated with celastrol (10 mg/kg dissolved in 1% DMSO + 2% Tween 80 + 97% water) for 5 consecutive days. After celastrol was treated for 3 days, ANIT and ANIT+Celastrol groups were given a single oral dose of ANIT. All mice were killed 48 h after ANIT administration. Liver samples were harvested and frozen at -80 °C before analysis. Results: A total of 978 DEGs were identified. Large numbers of these DEGs were related to activation of SIRT1, which included increased FXR signaling and inhibition of PPARγ, nuclear factor-kappa B (NF-κB), and P53 signaling. Conclusions: Celastrol could protect ANIT-induced cholestasis by recovering disrupted Sirt1 level.
Project description:Intrahepatic cholestasis (IC) is a common symptom of liver diseases but with limited treatment. Huangqi decoction (HQD), a classic herbal medicine, has shown protective effects against IC. In this study, the iTRAQ-based quantitative proteomics was performed to investigate the potential mechanism of HQD on alpha-naphthylisothiocyanate (ANIT) induced IC, resulting in 2,796 quantified proteins across all the samples, including 270 differentially expressed proteins under HQD treatment. Fuzzy c-means (FCM) clustering analysis of these 270 proteins revealed that the pro-inflammatory proteins, such as LCN2, SAA1, FGG, FGA, and FGB, were assigned in the Cluster 1 (up-regulated by ANIT, and down-regulated by HQD). Following functional bioinformatics analysis and protein-protein interaction (PPI) network analysis represented that these pro-inflammatory proteins were involved in the STAT3 signaling pathway. Further real-time PCR and Western blot experiments confirmed that the expression of these proteins was consistent with the proteomic results. Moreover, HQD treatment decreased the phosphorylation of STAT3 that induced by ANIT. And Western blot experiments also revealed that HQD could decrease phosphorylation of NF-κB and down-regulate the expression inflammatory genes IL-6, and therefore inhibit IL-6/STATA3 signaling pathway. In summary, the present study suggested that HQD treatment may ameliorate intrahepatic cholestasis via inhibiting the NF-κB/IL-6/STAT3 signaling pathway.
Project description:Celastrol plays a significant role in cerebral ischemia-reperfusion injury. Although previous studies have confirmed that celastrol post-treatment has a protective effect on ischemic stroke, the therapeutic effect of celastrol on ischemic stroke and the underlying molecular mechanism remain unclear. In the present study, focal transient cerebral ischemia was induced by transient middle cerebral artery occlusion (tMCAO) in mice and celastrol was administered immediately after reperfusion. We performed lncRNA and mRNA analysis in the ischemic hemisphere of adult mice with celastrol post-treatment through RNA-Sequencing (RNA-Seq). A total of 50 differentially expressed lncRNAs (DE lncRNAs) and 696 differentially expressed mRNAs (DE mRNAs) were identified between the sham and tMCAO group, and a total of 544 DE lncRNAs and 324 DE mRNAs were identified between the tMCAO and tMCAO+celastrol group. Bioinformatic analysis was done on the identified deregulated genes through gene ontology (GO) analysis, KEGG pathway analysis and network analysis. Pathway analysis indicated that inflammation-related signaling pathways played vital roles in the treatment of ischemic stroke by celastrol. Our study suggests celastrol treatment can effectively reduce cerebral ischemia-reperfusion injury. The bioinformatics analysis of lnRNAs and mRNAs profiles in the ischemic hemisphere of adult mice provides a new perspective in the neuroprotective effects of celastrol, particularly with regards to ischemic stroke.
Project description:Given that celastrol?s leptin-sensitizing effect requires high levels of circulating leptin, but lean mice have low levels of circulating leptin so that celastrol has no effect on lean mice. Analysis celastrol-induced hypothalamic gene expression profile change in lean mice will also be serving as negative control for DIO mice analysis.
Project description:The aim of the present study was to investigate the therapeutic mechanism of the anti-cholestatic effects of melatonin against ANIT-induced liver injury in rats, and to screen for potential biomarkers of cholestasis through isobaric tags for relative and absolute quantitation (iTRAQ) quantitative proteomics.
Project description:Given that celastrol?s leptin-sensitizing effect requires both high levels of circulating leptin and intact leptin receptor signaling, we analyzed the effect of celastrol on hypothalamic gene expression profile of db/db mice, which have high circulating levels of leptin, but lack intact leptin receptor signaling. This analysis will be serving as negative control for DIO mice analysis.
Project description:<p>Cholestasis is a clinical condition resulting from the impairment of bile flow. Currently, patients with cholestasis still face several barriers in seeking diagnosis and treatment. Zhuyu Pill (ZYP) is an ancient classic formula of the Coptis-Evodia herb couples (CEHC), and has been used for cholestasis treatment in the clinic, however, its underlying biological mechanism in cholestasis remain to be clarified. In this study, a cholestasis rat model, induced by α-naphthyl-isothiocyanate (ANIT, 50mg/kg) and treated with ZYP (0.6g/kg or 1.2g/kg), was adopted. Serum biochemical indices and histopathological evaluation was performed, in addition to metabolomics analyses of fecal and 16S rDNA sequencing of the gut microbiota. We evaluated the anticholestatic activity of ZYP and investigated the mechanism underlying its correlation with gut microbiota and fecal metabolite regulation. The relationships between biochemical indices, fecal metabolites, and gut microbiota were analyzed. The results showed that both high and low doses of ZYP can effectively improve the blood biochemical parameters of cholestasis rats, and the intervention effect of high dose ZYP is superior to that that of lower dose. Based on a metabolomics test of fecal samples, significantly altered metabolites in the ANIT and ZYP treatment group were identified. In total, 734 metabolites were differentially expressed, and whose biological functions were mainly associated with amino acid metabolism, steroid hormone biosynthesis, and bile secretion. In addition, sequencing of the 16S rDNA unit in fecal samples revealed that the ZYP could improve the gut microbiota dysbiosis that ANIT had induced. Therefore, we conclude that ANIT altering of blood biochemical and metabolic profiles and gut microbiota can be effectively alleviated using ZYP treatment, this study contributes “TCM wisdom” to clinical diagnosis and treatment of cholestasis.</p>
Project description:Purpose: This study aimed to explore the mechanism of Celastrol in improving psoriasis through single-cell transcriptomics Methods: Supplementation with intragastric administration of celastrol in C57BL/6 mice to observe its effect on IMQ-induced psoriasis. Single-cell RNA sequencing were performed to explore the role of celastrol for IMQ-induced psoriasis. Results: A natural product library was used to screen for a small molecule compound, celastrol, that could interfere with fibroblast-macrophage communication. It was demonstrated that celastrol targeted low-denisity lipoprotein receptor-related protein 1 (LRP1) to inhibit fibroblast secretion of CCL2 and inhibited psoriasis progression by reducing its recruitment to macrophages, thereby blocking communication between the two cells Conclusion: We report that celastrol targeted low-denisity lipoprotein receptor-related protein 1 (LRP1) to inhibit fibroblast secretion of CCL2 and inhibited psoriasis progression by reducing its recruitment to macrophages. The use of celastrol maybe a noveltherapeuticoption for psoriasis.
Project description:Since leptin signaling in the hypothalamus is critical to regulate food intake and body weight, we investigated how celastrol alters the hypothalamic transcriptome of DIO mice. By doing this analysis, genes with potential relevance for celastrol-mediated leptin sensitization could be identified.
Project description:Overexpression of p21 in NEMOM-NM-^Thepa animals protects against DNA damage, acceleration of hepatocarcinogenesis and cholestasis. As strengthened by our LPS stimulation experiments, we identified a novel protective role of p21 against DNA damage. Expression profiling of livers from wild type, NEMO, and NEMO-P21 null mice.