HPLC analysis. (A) HPLC chromatogram of standard compounds: 1, geniposidic acid; 2, deacetylasperulosidic acid; 3, genipin-1-gentiobioside; 4, geniposide; 5, Crocin I; and 6 and Crocin II. (B) HPLC chromatogram of GJE component: 1, geniposidic acid; 2, deacetylasperulosidic acid; 3, genipin-1-gentiobioside; and 4, geniposide were identified with a detection wavelength of 238 nm. (C) HPLC chromatogram of GJE component: 5, Crocin I; and 6, Crocin II were identified at a detection wavelength of 440 nm. HPLC, High-performance liquid chromatography; GJE, Gardenia jasminoides extracts.

Effects of LPS (A) and GJE (B) on cell viability. The specific concentration of LPS or GJE was used to treat ARPE-19 cells for 24 h, prior to MTT assay being used to measure cell viability, with untreated cells as a control. Data are presented as the mean ± standard error of the mean and analysed by one-way analysis of variance, followed by Dunnett's post-hoc test. **P<0.01, ***P<0.001. LPS, lipopolysaccharides; GJE, Gardenia jasminoides extracts.

Effects of GJE on the expression of proinflammatory cytokines. (A) mRNA levels of IL-1β, IL-6 and TNF-α in treated and untreated cells were measured by reverse transcription-quantitative polymerase chain reaction. (B) Secreted IL-1β, IL-6 and TNF-α protein levels in media of untreated and treated cells were measured using ELISA. Data are presented as the mean ± standard error of the mean and analyzed using one-way analysis of variance, followed by Tukey's post-hoc test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. GJE, Gardenia jasminoides extracts; IL, interleukin; TNF, tumor necrosis factor.

GJE inhibits CuSO4-induced primitive macrophage migration in transgenic zebrafish (Tg:zlyz-EGFP) embryos; magnification, x100. (A) Representative images of control and treated zebrafish embryos. (B) Macrophages in the regions of interest (white boxes) were quantified. Data are presented as the mean ± standard error of the mean and analysed using one-way analysis of variance, followed by Tukey's post-hoc test. *P<0.05, ****P<0.0001. GJE, Gardenia jasminoides extracts; IL, interleukin; TNF, tumour necrosis factor; LPS, lipopolysaccharides.

Effects of GJE on inflammation via the TLR4 signaling pathway in CuSO4-treated zebrafish embryos. (A) mRNA levels of IL-1β, IL-6 and TNF-α in treated and untreated embryos were measured by reverse transcription-quantitative polymerase chain reaction. (B) Expression levels of of NFKβ and TLR4 in untreated or treated embryos were measured by qRT-PCR. Data are presented as the mean ± standard error of the mean and analyzed using one-way analysis of variance, followed by Tukey's post-hoc test. *P<0.05, **P<0.01. GJE, Gardenia jasminoides extracts; TLR4, toll-like receptor 4; ns, no significance; IL, interleukin; TNF, tumor necrosis factor; LPS, lipopolysaccharides.

Effects of GJE on inflammation via the TLR4 signaling pathway in LPS-treated zebrafish embryos. GJE decreased LPS-induced expression of proinflammatory cytokine genes (A), attenuated LPS-induced expression of NFKβ activating protein and decreased LPS-induced TLR4 expression (B). Data are presented as the mean ± standard error of the mean and analyzed using one-way analysis of variance, followed by Tukey's post-hoc test. *P<0.05, **P<0.01. GJE, Gardenia jasminoides extracts; TLR4, toll-like receptor 4; ns, no significance; IL, interleukin; TNF, tumor necrosis factor; LPS, lipopolysaccharides.