Celastrol is a novel selective agonist of cannabinoid receptor 2 with anti-inflammatory and anti-fibrotic activity in a mouse model of systemic sclerosis
Celastrol is a novel selective agonist of cannabinoid receptor 2 with anti-inflammatory and anti-fibrotic activity in a mouse model of systemic sclerosis
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DOI：
10.1016/j.phymed.2019.153160
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发表时间：
2020-02-01
期刊：
影响因子：
7.9
通讯作者：
Lu, Weiqiang
中科院分区：
文献类型：
--
作者：
Jiang, Xingwu;Chen, Si;Lu, Weiqiang
背景：越来越多的证据表明大麻素受体参与了器官纤维化的发病过程。目的：本研究的目的是寻找新的大麻素受体2(CB2)激动剂，并评价其在治疗系统性硬化症中的潜力。方法：建立了一种基于赤子荧光素酶的裂解荧光素酶互补试验(SLCA)来检测CB2与β-抑制素2的相互作用。然后用SLCA方法筛选出366个天然产物的文库，作为潜在的CB2激动剂。几种GPCR功能检测，包括基于hTRF的cAMP检测和钙动员，也被用来评估CB2的激活。采用博莱霉素诱导的实验性系统性硬化症模型评价其体内抗肝纤维化作用。通过H&E和天狼星红染色法测定真皮厚度和胶原含量。结果：采用SLCA法鉴定雷公藤红素为CB2的新激动剂。此外，雷公藤红素以剂量依赖的方式触发CB2介导的多个下游信号通路，包括钙动员、抑制cAMP积累和受体脱敏，并且对CB1具有适度的选择性。此外，雷公藤红素对脂多糖处理的小鼠原始264.7巨噬细胞和原代巨噬细胞具有抗炎作用。结论：雷公藤红素是一种新型的选择性CB2激动剂。雷公藤红素激活CB2可减轻炎症反应，减少炎症反应。结论：雷公藤红素是一种新型的选择性CB2激动剂。
Background: Increasing evidence indicated that the cannabinoid receptors were involved in the pathogenesis of organ fibrogenesis.Purpose: The purpose of this study was to discover novel cannabinoid receptor 2 (CB2) agonist and assess the potential of CB2 activation in treating systemic sclerosis.Methods: A gaussia princeps luciferase-based split luciferase complementation assay (SLCA) was developed for detection of the interaction between CB2 and beta-arrestin2. A library of 366 natural products was then screened as potential CB2 agonist using SLCA approach. Several GPCR functional assays, including HTRF-based cAMP assay and calcium mobilization were also utilized to evaluated CB2 activation. Bleomycin-induced experimental systemic sclerosis was used to assess the in vivo anti-fibrotic effects. Dermal thickness and collagen content were evaluated via H&E and sirius red staining.Results: Celastrol was identified as a new agonist of CB2 by using SLCA. Furthermore, celastrol triggers several CB2-mediated downstream signaling pathways, including calcium mobilization, inhibition of cAMP accumulation, and receptor desensitization in a dose-dependent manner, and it has a moderate selectivity on CB1. In addition, celastrol exhibited the anti-inflammatory properties on lipopolysaccharide (LPS) treated murine Raw 264.7 macrophages and primary macrophages. Finally, we found that celastrol exerts anti-fibrotic effects in the bleomycin-induced systemic sclerosis mouse model accompanied by reduced inflammatory conditions.Conclusion: Taken together, celastrol is identified a novel selective CB2 agonist using a new developed arrestin-based SLCA, and CB2 activation by celastrol reduces the inflammatory response, and prevents the development of dermal fibrosis in bleomycin-induced systemic sclerosis mouse model.