GTE treatment on cell infiltration and protein exudation in aqueous humor.

<p>(A) and (B): GTE caused a significant reduction in accumulation of cells (A) and protein exudation (B) in aqueous humor (***<i>p</i><0.01) when compared with LPS+water. The differences among the GTE treatment groups were, however, not significant (<i>p</i>>0.05). Saline+water, n = 3; LPS+water, n = 6; LPS+GTE1, n = 6; LPS+GTE2, n = 6; LPS+GTE4, n = 6; LPS+Dxm, n = 6; Saline+water, n = 3. (C) and (D): In another experiment, significant reduction in infiltrating cells (C) and protein content (D) in aqueous humor was observed only in rats treated with 550 mg/kg GTE (*<i>p</i><0.05), but not in those received half dose (275 mg/kg) of GTE. Saline+water, n = 5; n = 6 in LPS+water, LPS+GTE2 550 and LPS+GTE2 275. Data were shown as mean ± <i>SE</i>.</p

Effect of GTE on macrophages within iris and ciliary body.

<p>(A)–(D): Fluorescent micrographs showing some cells were immunopositive to CD68, a marker for macrophages (arrows, magnified in inserts) 24 hours after treatment with (A): saline+water, (B): LPS+water, (C): LPS+GTE4, (D): LPS+Dxm. (E): A significant reduction in number of macrophages presented in the stroma of iris and ciliary body was observed after treatment with GTE and dexamethasone (Dxm). n = 3 in each group. Data were shown as mean ± <i>SE</i>. **<i>p</i><0.05 compared with LPS+water. C: cornea; AC: anterior chamber; I: iris; PC: posterior chamber; L: lens; CB: ciliary body.</p

CD14 and TLR-4 mRNA expression in the iris and ciliary body and the retina by quantitative real-time PCR analyses.

<p>C_T: threshold cycle; Saline+water: saline injection and fed with water; LPS+water: LPS injection and fed with water; LPS+GTE1: GTE was fed once after LPS injection; LPS+GTE2: GTE was fed twice after LPS injection; LPS+GTE4: GTE was fed four times after LPS injection; LPS+Dxm: Dxm was fed once after LPS injection; Saline+GTE4: saline injection and fed as GTE4.</p

Primer sequences used for quantitative real-time PCR detection of rat-CD14, TLR-4 and GAPDH.

<p>CD14: cluster of differentiation 14; TLR-4: toll-like receptor 4; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.</p

Clinical manifestations of ocular inflammation in rat eyes.

<p>Ocular inflammation was evaluated by slit lamp examination 24 hours after LPS injection. (A): No inflammatory feature was observed in normal control rats (saline+water). (B) and (C): Hyperemia (green arrow), edema (yellow arrow) and synachesia (purple arrow) occurred in the iris of LPS treated rats. (D) and (E): Inflammatory responses were subsided in rats treated with GTE1 (D) and GTE4 (E). (F): Inflammatory responses were also suppressed in rats treated with Dexamethasone (Dxm). (G): The scores of clinical features were reduced significantly after GTE and Dxm treatments (**<i>p<</i>0.05, when compared with LPS+water). Saline+water, n = 3; LPS+water, n = 6; LPS+GTE1, n = 6; LPS+GTE2, n = 6; LPS+GTE4, n = 6; LPS+Dxm, n = 6. Data were shown as mean ± <i>SE</i>. Scar bar = 2 mm.</p

TNF-α, IL-6 and MCP-1 concentration in the serum after different treatments.

<p>-: undetectable. Sensitivity of these assays is 5 pg/mL for TNF-α, 21 pg/mL for IL-6, less than 8.0 pg/mL for MCP-1.</p><p>Saline+water: saline injection and fed with water; LPS+water: LPS injection and fed with water; LPS+GTE1: GTE was fed once after LPS injection; LPS+GTE2: GTE was fed twice after LPS injection; LPS+GTE4: GTE was fed four times after LPS injection; LPS+Dxm: Dxm was fed once after LPS injection; Saline+GTE4: saline injection and fed as GTE4; **<i>p</i><0.05, compared with LPS+water.</p

Effects of GTE on cytokine and chemokine production in aqueous humor.

<p>The pro-inflammatory factors TNF-α (A), IL-6 (B) and MCP-1 (C) were all reduced significantly after GTE treatments when compared with LPS+water (***<i>p<</i>0.01). Within group comparisons, however, did not show obvious differences among GTE treated groups (<i>p</i>>0.05). Sensitivity of the assays: 5 pg/mL for TNF-α, 21 pg/mL for IL-6, less than 8.0 pg/mL for MCP-1. Saline+water, n = 3; LPS+water, n = 6; LPS+GTE1, n = 6; LPS+GTE2, n = 6; LPS+GTE4, n = 6; LPS+Dxm, n = 6; Saline+water, n = 3. Data were shown as mean ± <i>SE</i>.</p

Histological features of infiltrating cells in ocular tissues.

<p>Paraffin sections showed infiltrating cells in the anterior segments of the eye after LPS insult. (A): H&E section showed clusters of infiltrating cells (arrow, magnified in the insert) with polymorphic nuclei in anterior (AC) and posterior chamber (PC). (B): Giemsa staining of infiltrating cells (arrows) in aqueous humor. (D) and (E): Fluorescent micrographs showing localization of CD43 on leukocytes (arrow, magnified in the insert) in iris (I), PC and AC. (G) and (H): CD68 was localized on macrophage/monocytes (arrows, magnified in the insert) in iris, PC and AC. No staining was observed in control sections treated without primary antibody against CD43 (C) or CD68 (F). C: cornea.</p

The number of infiltrating cells in ocular tissues after GTE treatments.

<p>(A), (B), (C) and (D): H&E sections showing accumulation of infiltrating cells (orange arrows, magnified in inserts) and protein exudation (green arrow) in anterior segments of the eye. (E), (F), (G) and (H): H&E sections showing infiltrating cells (arrows) in vitreous body (VB) around the optic nerve head (ONH). (A, E): Saline+water; (B, F): LPS+water; (C, G): LPS+GTE4; (D, H): LPS+Dxm. (I): The number of infiltrating cells in the anterior and posterior segments of the eye was reduced in rats treated with GTE (**<i>p</i><0.05) when compared with that with LPS and vehicle treatment. n = 3 in each group. Data were shown as mean ± <i>SE</i>. L: lens; CB: ciliary body; R: retina.</p