Anti-inflammatory Mechanism of Geniposide: Inhibiting the Hyperpermeability of Fibroblast-Like Synoviocytes via the RhoA/p38MAPK/NF-κB/F-Actin Signal Pathway
Anti-inflammatory Mechanism of Geniposide: Inhibiting the Hyperpermeability of Fibroblast-Like Synoviocytes via the RhoA/p38MAPK/NF-κB/F-Actin Signal Pathway
复制标题
京尼平苷的抗炎机制：通过RhoA/p38MAPK/NF-kB/F-Actin信号通路抑制成纤维样滑膜细胞的高通透性
DOI：
10.3389/fphar.2018.00105
复制
发表时间：
2018-02-15
影响因子：
5.6
通讯作者：
Fu, Jun
中科院分区：
文献类型：
--
作者：
Deng, Ran;Li, Feng;Fu, Jun
栀子苷（Geniposide， GE）是从栀子中提取纯化的环烯醚萜苷类化合物，是一种很有前途的抗炎药物，但其对类风湿关节炎（RA）的作用机制尚未明确。本研究探讨GE降低SD大鼠佐剂性关节炎（AA）成纤维细胞样滑膜细胞（FLSs）高通透性的分子机制，旨在观察GE对AA大鼠的作用，探索治疗RA的新策略。CCK-8法检测FLSs增殖。采用酶联免疫吸附试验（ELISA）试剂盒检测FLSs的促炎细胞因子和抗炎细胞因子水平。在Transwell上进行FLSs渗透率测定。免疫荧光法检测f -肌动蛋白的排列和形态。western blotting检测与FLSs通透性相关的关键分子（RhoA、p-p38MAPK、NF-kappa B、p-p65和F-actin）的表达。脂多糖（LPS）处理后，细胞的增殖能力和通透性均显著提高（P < 0.05）。与对照组相比，FLSs中RhoA、p-p38MAPK、NF-kappa B p-p65和F-actin的表达增加，F-actin重新分布，形成额外的应力纤维。但是，这些情况在GE治疗后得到缓解。我们证明了不同浓度的GE（25、50和100 mu g/mL）处理对体外FLSs的增殖和通透性有显著的抑制作用。此外，不同剂量GE组FLSs分泌的白细胞介素(IL)-1 β和IL-17水平降低，抗炎细胞因子（IL-4、tgf - β 1）水平升高。GE处理下，低表达的RhoA较对照组下调了p-p38MAPK、NF-kappa B、p-p65、F-actin的表达，恢复了LPS处理导致的FLSs高通透性。综上所述，GE可能通过调节促炎细胞因子和抗炎细胞因子的相对平衡来发挥其抗炎和免疫调节作用。GE减弱了FLSs的超通透性。RhoA/p38MAPK/NF-kappa B/F-actin信号传导的下调可能在GE对RA的作用机制中起关键作用。GE可能是治疗类风湿性关节炎的有效药物。
Geniposide (GE) is the extraction and purification of iridoid glycosides from the Gardenia jasminoides Ellis, which is a promising anti-inflammatory drug, but its mechanism of actions on rheumatoid arthritis (RA) has not been clarified. This study investigated the molecular mechanism behind GE reduced the high permeability of fibroblast-like synoviocytes (FLSs) derived from SD rats with adjuvant arthritis (AA), with the aims of observing the action of GE in AA rats and exploring new therapeutic strategies for RA treatment. The CCK-8 method was used to detect FLSs proliferation. The pro-inflammatory cytokines levels and anti-inflammatory cytokines levels in FLSs were determined by ELISA kits. FLSs permeability assay was performed on Transwell. Immunofluorescence was used to assay the arrangement and morphology of F-actin. The expression of the key molecules related to FLSs permeability (RhoA, p-p38MAPK, NF-kappa B p-p65 and F-actin) was detected by western blotting. After treatment with lipopolysaccharide (LPS), the proliferation and the permeability of the cells increased significantly (all P < 0.05). The expression of RhoA, p-p38MAPK, NF-kappa B p-p65 and F-actin in FLSs was higher compared with the control group, and F-actin was redistributed, with the formation of additional stress fibers. But, these conditions were moderated after treatment with GE. We demonstrated that the treatment of different concentrations of GE (25, 50, and 100 mu g/mL) had a significant inhibitory effect on the proliferation and permeability of FLSs in vitro. Furthermore, the levels of interleukin (IL)-1 beta and IL-17 secreted by FLSs were decreased in different doses of GE groups, and the levels of anti-inflammatory cytokines (IL-4, TGF-beta 1) were increased. Under treatment with GE, low expression of RhoA downregulated expression of p-p38MAPK, NF-kappa B p-p65, and F-actin while compared with control group, and restored the hyperpermeability of FLSs due to LPS treatment. Taken together, GE might play its anti-inflammatory and immunoregulatory effects via regulating the relative equilibrium of pro-inflammatory cytokines and anti-inflammatory cytokines. GE attenuated the hyperpermeability of FLSs. The down-regulation of the conduction of RhoA/p38MAPK/NF-kappa B/F-actin signal may play a critical role in the mechanisms of GE on RA. GE could be an effective therapeutic agent for the treatment of RA.