Sulforaphane elicts dual therapeutic effects on Renal Inflammatory Injury and crystal deposition in Calcium Oxalate Nephrocalcinosis
Sulforaphane elicts dual therapeutic effects on Renal Inflammatory Injury and crystal deposition in Calcium Oxalate Nephrocalcinosis
复制标题
萝卜硫素对肾脏炎症损伤和草酸钙肾钙质沉着症中的晶体沉积具有双重治疗作用。
DOI：
10.7150/thno.44054
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Xu, Hua
中科院分区：
文献类型：
--
作者：
Liu, Haoran;Yang, Xiaoqi;Xu, Hua
肾内草酸钙（CaOx）晶体可诱导肾小管上皮细胞（TEC）损伤和炎症，这与Toll样受体4（TLR 4）/干扰素调节因子1（IRF 1）信号通路有关。此外，浸润的巨噬细胞（M φ s）可能会影响肾内CaOx晶体和CaOx诱导的肾损伤。尽管核因子红细胞2相关因子2（Nrf 2）在调节炎症和巨噬细胞极化中的作用已得到充分表征，但其调节CaOx肾钙沉着症的潜在机制仍不确定。方法：我们使用基因表达综合数据集来分析基因表达谱。采用荧光素酶报告基因、western blot、定量聚合酶链反应、免疫荧光染色、荧光原位杂交、正电子发射计算机断层扫描成像、流式细胞术和染色质免疫沉淀法研究Nrf 2调控miR-93-TLR 4/IRF 1的机制。抗炎活性和调节的巨噬细胞极化Nrf 2在体外和体内investigatedinvivo.Results：我们发现，结石介导的肾脏炎症显着影响结石的生长，萝卜硫素衰减CaOx肾钙化引起的肾损伤和肾CaOx晶体沉积。此外，在萝卜硫素治疗后的CaOx肾钙质沉着症小鼠模型中，Nrf 2水平显著增加，并与TLR 4和IRF 1水平呈负相关。此外，Nrf 2抑制TLR 4和IRF 1的水平，并减少由COM刺激的TEC的上清液诱导的M1-巨噬细胞极化。在机制方面，转录因子分析、microRNA微阵列和染色质免疫沉淀分析表明，Nrf 2表现出对miR-93- 5 p的正转录激活。此外，荧光素酶报告基因、qRT-PCR和western blot验证了miR-93- 5 p靶向TLR 4和IRF 1 mRNA。结论：萝卜硫素可能通过抑制Nrf 2- miR-93-TLR 4/IRF 1通路，促进M2 M phi极化，抑制CaOx肾钙蛋白沉积诱导的肾小管上皮细胞炎症损伤。
Intrarenal calcium oxalate (CaOx) crystals induce renal tubular epithelial cells (TECs) injury and inflammation, which involve Toll-like receptor 4 (TLR4)/interferon regulatory factor 1 (IRF1) signaling. Additionally, infiltrating macrophages (M phi s) might influence intrarenal CaOx crystals and CaOx-induced renal injury. Although the roles of nuclear factor erythroid 2-related factor 2 (Nrf2) in regulating inflammation and macrophage polarization are well characterized, its potential mechanisms in regulating CaOx nephrocalcinosis remain undefined.Methods: We used a Gene Expression Omnibus dataset to analyze gene-expression profiles. Luciferase reporter, western blot, quantitative polymerase chain reaction, immunofluorescence staining, fluorescence in situ hybridization, positron emission tomography computed tomography imaging, flow cytometry, and chromatin immunoprecipitation assays were employed to study the mechanism of miR-93-TLR4/IRF1 regulation by Nrf2. Anti-inflammatory activity and regulation of macrophage polarization by Nrf2 were investigated in vitro and in vivo .Results: We found that stone-mediated kidney inflammation significantly affected stone growth, and that sulforaphane attenuated CaOx nephrocalcinosis-induced kidney injury and renal CaOx crystals deposition. Additionally, Nrf2 levels significantly increased and negatively correlated with TLR4 and IRF1 levels in a mouse model of CaOx nephrocalcinosis following sulforaphane treatment. Moreover, Nrf2 suppressed TLR4 and IRF1 levels and decreased M1-macrophage polarization which induced by supernatants from COM-stimulated TECs in vitro . In terms of mechanism, transcription factor analyses, microRNA microarray, and chromatin immunoprecipitation assays showed that Nrf2 exhibited positive transcriptional activation of miR-93-5p. In addition, Luciferase reporter, qRT-PCR, and western blot validated that miR-93-5p targets TLR4 and IRF1 mRNA. Furthermore, suppressed miR-93-5p expression partially reversed Nrf2-dependent TLR4/IRF1 downregulation.Conclusions: The results suggested that sulforaphane might promote M2M phi polarization and inhibit CaOx nephrocalcinosis-induced inflammatory injury to renal tubular epithelial cells via the Nrf2- miR-93-TLR4/IRF1 pathway in vitro and in vivo.