Department of Pharmacology, School of Pharmacy, Nanjing University of Chinese Medicine, 138 Xianlin Avenue, Nanjing, Jiangsu 210023, China.
Department of Pharmacology, School of Pharmacy, Nanjing University of Chinese Medicine, 138 Xianlin Avenue, Nanjing, Jiangsu 210023, China; Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica, Nanjing University of Chinese Medicine, China; Jiangsu Key Laboratory of Therapeutic Material of Chinese Medicine, Nanjing University of Chinese Medicine, China.
Toxicology. 2020 Jul;440:152475. doi: 10.1016/j.tox.2020.152475. Epub 2020 Apr 25.
Curcumol, a guaiane-type sesquiterpenoid hemiketal extracted from the herb Rhizoma Curcumae, exhibits multiple-pharmacological activities. We previously reported that curcumol ameliorated hepatic fibrosis by inhibiting hepatic stellate cell (HSC) activation. In this study, we aimed to investigate the effect of curcumol on HSC migration and adhesion, and reveal its regulation mechanisms.
Cellular viability was determined by Cell Counting Kit-8. Cell migration was detected by boyden chamber and cell scratch experiment. Recombinant human periostin (rh POSTN) and adeno-associated viral (AAV)-GFP-periostin were used to achieve POSTN overexpression in vitro and in vivo, respectively. Nuclear factor kappa B (NF-κB)-p65 overexpression was achieved by using plasmid. ELISA was conducted to detect POSTN level. Immunohistochemistry, qRT-PCR, Western blotting, and immunofluorescence were performed to assess associated factor expression.
Curcumol suppressed HSC migration and adhesion, and reduced the secretion and expression of POSTN. By gain of function POSTN in HSCs, using rh POSTN, we found that the inhibition of HSC migration and adhesion by curcumol depended on the decrease of POSTN. Besides, curcumol protection against chronic CCl-caused hepatic fibrosis could be impaired by POSTN overexpression. Moreover, we showed that curcumol repressed NF-κB signaling and the production of pro-inflammatory factor. Importantly, curcumol down-regulation of POSTN was rescued by knock-in of NF-κB, as well as the inhibition of HSC migration and adhesion.
These findings reveal the molecular mechanism of curcumol-reduced HSC migration and adhesion, by which points to the possibility of using curcumol based on NF-κB dependent POSTN for the treatment of fibrogenesis.
莪术醇是从姜黄属植物根茎中提取的一种愈创木烷型倍半萜半缩醛，具有多种药理活性。我们之前的研究表明，莪术醇通过抑制肝星状细胞（HSC）活化来改善肝纤维化。在这项研究中，我们旨在研究莪术醇对 HSC 迁移和黏附的影响，并揭示其调控机制。
用细胞计数试剂盒-8 测定细胞活力。通过 Boyden 室和细胞划痕实验检测细胞迁移。重组人骨桥蛋白（rh POSTN）和腺相关病毒（AAV）-GFP-骨桥蛋白分别用于体外和体内实现 POSTN 的过表达。核因子 kappa B（NF-κB）-p65 过表达通过质粒实现。ELISA 用于检测 POSTN 水平。免疫组织化学、qRT-PCR、Western blot 和免疫荧光用于评估相关因子的表达。
莪术醇抑制 HSC 迁移和黏附，并降低 POSTN 的分泌和表达。通过在 HSCs 中过表达 POSTN，使用 rh POSTN，我们发现莪术醇抑制 HSC 迁移和黏附依赖于 POSTN 的减少。此外，POSTN 的过表达可损害莪术醇对慢性 CCl 引起的肝纤维化的保护作用。此外，我们表明莪术醇抑制 NF-κB 信号和促炎因子的产生。重要的是，NF-κB 的敲入以及 HSC 迁移和黏附的抑制挽救了莪术醇下调 POSTN。
这些发现揭示了莪术醇减少 HSC 迁移和黏附的分子机制，提示基于 NF-κB 依赖的 POSTN 的莪术醇治疗纤维化的可能性。
