Anti-inflammatory Mechanism of Geniposide: Inhibiting the Hyperpermeability of Fibroblast-Like Synoviocytes via the RhoA/p38MAPK/NF-κB/F-Actin Signal Pathway

Abstract

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-κB 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-κB 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 μg/mL) had a significant inhibitory effect on the proliferation and permeability of FLSs in vitro. Furthermore, the levels of interleukin (IL)-1β and IL-17 secreted by FLSs were decreased in different doses of GE groups, and the levels of anti-inflammatory cytokines (IL-4, TGF-β1) were increased. Under treatment with GE, low expression of RhoA downregulated expression of p-p38MAPK, NF-κB 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-κB/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.

mRNA level of ROCK1, RHOA, and LIMK2 as genes associated with apoptosis in evaluation of effectiveness of adalimumab treatment

“…The overexpression of these three factors was described in rheumatoid arthritis, which indicates that their inhibition would be a promising new therapeutic target [41][42][43]. Moreover, an analysis that was performed by Deng et al confirmed the important contribution of RhoA in cartilage destruction and the metabolism of synoviocytes [44]. Also, Kumar et al and Cho et al confirmed the role of RhoA in mediating the process of the formation of a network of new blood vessels, as well as with the migration of epithelial cells, which were mediated by the TGFβ and E-cadherin pathway [45,46].…”

Re-Du-Ning inhalation solution exerts suppressive effect on the secretion of inflammatory mediators via inhibiting IKKα/β/IκBα/NF-κB, MAPKs/AP-1, and TBK1/IRF3 signaling pathways in lipopolysaccharide stimulated RAW 264.7 macrophages

“…32 Moreover, geniposide also exerted potential anti-inammatory activity via inhibition of the NF-kB signaling pathway. 33 Hence, the suppressive effects of RIS on inammatory mediators may be, at least in part, due to the presence of these two compounds. Further studies will be conducted to identify other active components responsible for the anti-inammatory effect of RIS.…”

Herbal compounds for rheumatoid arthritis: Literatures review and cheminformatics prediction

“…Some previous researches have shown that betulinic acid, kaempferol, leonurine, oxymatrine, and piperlongumine can inhibit the migration and invasion of FLS in RA models (Li et al, ; Liang et al, ; Liu, Feng, et al, ; Pan et al, ; Sun, Xu, Du, Zhang, & Zhu, ). Triptolide, celastrol, geniposide, paeoniflorin, clematichinenoside AR, glycyrrhetinic acid, kirenol, cucurbitacin E, 1,7‐dihydroxyl‐3,4‐dimethoxylxanthone, calycosin, catechin, cyanidin‐3‐glucoside, quercetin, α‐mangostin, leonurine, norisoboldine, oxymatrine, caffeic acid, paeonol, resveratrol, shikonin, and 3 R ‐(4′‐hydroxyl‐3′‐ O ‐β‐ d ‐glucopyranosyl phenyl)‐dihydro isocoumarin alleviate inflammatory response by inhibiting the release of a variety of proinflammatory factors (e.g., activin A, ciliary neurotrophic factor, and fractalkine) in FLS of RA models (Bi et al, ; Chen et al, ; Deng et al, ; Huang et al, ; Jia et al, ; Li et al, ; Liang et al, ; Lin, Sato, & Ito, ; Liu, Feng, et al, ; Mao, Sun, Pei, & Zhang, ; Su et al, ; Su et al, ; Sun et al, ; Sun et al, ; Sun & Li, ; Sung et al, ; Tang et al, ; Tian, Chen, Gao, Li, & Xie, ; Venkatesha, Astry, Nanjundaiah, Yu, & Moudgil, ; Wang, Sun, & Jin, ; Wei et al, ; Yang et al, ; Yang et al, ; Zhang et al, ; Zheng et al, ; Zuo et al, ).…”

“…In addition to inflammatory damage, FLS also plays an essential role in cartilage destruction (Pan et al, ). Therefore, the decrease in FLS activity by some components is also beneficial to the protection of cartilage in RA models, including triptolide, celastrol, geniposide, paeoniflorin, clematichinenoside AR, kirenol, tanshinone IIA, bauchampine A, betulinic acid, baicalein, 1,7‐dihydroxyl‐3,4‐dimethoxylxanthone, catechin, kaempferol, licochalcone A, luteolin, quercetin, ramosissimin, α‐mangostin, berberine, fissistigmine A, matrine, norisoboldine, oxymatrine, piperlongumine, sinomenine, tamaractam, caffeic acid, chlorogenic acid, paeonol, daphnetin, and leonurine (Bi et al, ; Chen et al, ; Chen et al, ; Deng et al, ; Hong et al, ; Hua et al, ; Jie et al, ; Kusunoki et al, ; Li et al, ; Li et al, ; Li et al, ; Liang et al, ; Liu et al, ; Liu, Feng, et al, ; Lou et al, ; Lou et al, ; Luo et al, ; Pan et al, ; Pan et al, ; Shu et al, ; Su et al, ; Su et al, ; Sun et al, ; Sun et al, ; Sun et al, ; Sung et al, ; Tang et al, ; Wang et al, ; Wang, Dai, et al, ; Wang, Sun, & Jin, ; Xu et al, ; Yang, Dong, & Li, ; Yao et al, ; Yi et al, ; Zhang et al, ; Zhang, Chen, et al, ; Zheng et al, ; Zhou et al, ; Zuo et al, ).…”