Chen Xiao-Wen, Liu Wen-Ting, Wang Yu-Xian, Chen Wen-Jing, Li Hong-Yu, Chen Yi-Hua, Du Xiao-Yan, Peng Fen-Fen, Zhou Wei-Dong, Xu Zhao-Zhong, Long Hai-Bo
Department of Nephrology, ZhuJiang Hospital, Southern Medical University, Guangzhou, 510280, China.
Department of Gerontology, ZhuJiang Hospital, Southern Medical University, Guangzhou, 510280, China.
J Diabetes Complications. 2016 Jul;30(5):760-9. doi: 10.1016/j.jdiacomp.2016.03.013. Epub 2016 Mar 17.
The aim of this research was to investigate the effects of cyclopropanyldehydrocostunolide (also named LJ), a derivative of sesquiterpene lactones (SLs), on high glucose (HG)-induced podocyte injury and the associated molecular mechanisms.
Differentiated mouse podocytes were incubated in different treatments. The migration and albumin filtration of podocytes were examined by Transwell filters. The protein and mRNA levels of MCP-1 were measured using enzyme-linked immunosorbent assay (ELISA) and quantitative real-time PCR (q-PCR). Protein expression and phosphorylation were detected by western blot, and the nuclear translocation of NF-κB was performed with a confocal microscope. The gene expression of the receptor activator for NF-κB (RANK) was silenced by small interfering RNA (siRNA).
Our results showed that HG enhanced migration, albumin filtration and MCP-1 expression in podocytes. At the molecular level, HG promoted the phosphorylation of NF-κB/p65, IKKβ, IκBα, mitogen-activated protein kinase (MAPK) and the nuclear translocation of p65. LJ reversed the effects of HG in a dose-dependent manner. Furthermore, our data provided the first demonstration that the receptor activator for NF-κB ligand (RANKL) and its cognate receptor RANK were overexpressed in HG-induced podocytes and were downregulated by LJ. RANK siRNA also attenuated HG-induced podocyte injury and markedly inhibited the activation of NF-κB and MAPK signaling pathways.
LJ attenuates HG-induced podocyte injury by suppressing RANKL/RANK-mediated NF-κB and MAPK signaling pathways.
本研究旨在探讨倍半萜内酯（SLs）衍生物环丙烷基脱氢木香内酯（又称LJ）对高糖（HG）诱导的足细胞损伤及相关分子机制的影响。
将分化的小鼠足细胞进行不同处理。通过Transwell小室检测足细胞的迁移和白蛋白滤过情况。采用酶联免疫吸附测定（ELISA）和定量实时聚合酶链反应（q-PCR）检测单核细胞趋化蛋白-1（MCP-1）的蛋白和mRNA水平。通过蛋白质印迹法检测蛋白表达和磷酸化情况，并用共聚焦显微镜检测核因子κB（NF-κB）的核转位。用小干扰RNA（siRNA）沉默核因子κB受体激活剂（RANK）的基因表达。
我们的结果表明，HG增强了足细胞的迁移、白蛋白滤过和MCP-1表达。在分子水平上，HG促进了NF-κB/p65、IκB激酶β（IKKβ）、IκBα、丝裂原活化蛋白激酶（MAPK）的磷酸化以及p65的核转位。LJ以剂量依赖的方式逆转了HG的作用。此外，我们的数据首次证明，在HG诱导的足细胞中，核因子κB受体激活剂配体（RANKL）及其同源受体RANK过表达，而LJ可使其下调。RANK siRNA也减轻了HG诱导的足细胞损伤，并显著抑制了NF-κB和MAPK信号通路的激活。
LJ通过抑制RANKL/RANK介导的NF-κB和MAPK信号通路减轻HG诱导的足细胞损伤。
