Xue-Yun Zhang

Effects of baicalin, baicalein, and wogonin on interleukin-6 and interleukin-8 expression, and nuclear factor-κb binding activities induced by interleukin-1β in human retinal pigment epithelial cell line

The objective of the study was to investigate the effects of baicalin, baicalein, and wogonin (plant flavonoids) on interleukin-6 (IL-6) and interleukin-8 (IL-8) protein production, mRNA expression, and nuclear factor-kappaB (NF-kappaB) binding activities induced by interleukin-1beta (IL-1beta) in human retinal pigment epithelial cell line (ARPE-19) cells. To induce IL-6 and IL-8 mRNA expression and protein levels, IL-1beta was added to serum-free medium of ARPE-19 cells and incubated. The flavonoids were added to the medium. IL-6 and IL-8 in the media were measured using enzyme-linked immunosorbent assay. Both IL-6 and IL-8 mRNA were measured by semiquantitative reverse transcription polymerase chain reaction. The binding activities of the transcription factor NF-kappaB complexes to IL-6 and IL-8 were measured by electrophoretic mobility shift assay. IL-6 and IL-8 in the culture media of ARPE-19 cells were increased by IL-1beta in a dose-dependent manner. Baicalin did not suppress IL-1beta-induced IL-6 and IL-8 production, but dexamethasone, baicalein, and wogonin, significantly suppressed IL-6 and IL-8 production. Elevation of IL-6 and IL-8 mRNA was not suppressed by baicalin but was significantly suppressed by dexamethasone, baicalein, and wogonin. NF-kappaB binding activities were not suppressed by baicalin and baicalein, but was suppressed by wogonin. Wogonin and baicalein inhibited IL-1beta-induced IL-6 and IL-8 mRNA and protein production in ARPE-19 cells. The data suggest that wogonin may inhibit IL-6 and IL-8 mRNA expression via the suppression of NF-kappaB binding activities.

Effect of Berberine on Barrier Function in a Human Retinal Pigment Epithelial Cell Line

PurposeTo examine the effects of berberine, an alkaloid isolated from some medicinal herbs, on the disruption of the barrier function in a human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1β (IL-1β).MethodsARPE-19 cells were cultured to confluence. Berberine and IL-1β were added to the medium. Barrier functions were evaluated by measuring transepithelial electrical resistance (TER) and the permeability to horseradish peroxidase (HRP) and sodium fluorescein (SF).ResultsBerberine dose-dependently inhibited decreased TER and increased the permeability to HRP and SF in the cells stimulated with IL-1β.ConclusionsBerberine dose-dependently inhibited the disruption of the barrier function in the ARPE-19 cell line induced by IL-1β. Jpn J Ophthalmol 2007;51:64–67

Effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on IL-6, IL-8, and MCP-1 expression in human retinal pigment epithelial cell line.

Purpose: To examine pituitary adenylate cyclase–activating polypeptide (PACAP) receptors (PAC1, VPAC1, and VPAC2) mRNA and the effect of PACAP on interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1) expression in human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1β (IL-1β). Methods: Expression of PACAP receptor mRNA was examined by reverse transcription polymerase chain reaction (RT-PCR). PACAP and IL-1β were added to serum-free medium. IL-6, IL-8, and MCP-1 mRNA were measured by real-time PCR. IL-6, IL-8, and MCP-1 protein concentrations were measured using enzyme-linked immunosorbent assay. Nuclear factor κB (NF-κB) translocation was examined by immunofluorescence. Results: PAC1 and VCAP1 receptors mRNA were expressed in unstimulated cells. VCAP2 mRNA was expressed in cells stimulated with IL-1β. IL-1β stimulated IL-6, IL-8, and MCP-1 mRNA expression and protein levels. PACAP (10− 7 to 10− 6 M) inhibited IL-1β –stimulated IL-6, IL-8, and MCP-1 mRNA and protein levels. Immunofluorescence of NF-κB in the nucleus was dense 30 min after stimulation with IL-1β, and it was decreased by PACAP. Conclusions: ARPE-19 cells had PACAP receptors mRNA. PACAP inhibited IL-6, IL-8, and MCP-1 expression and protein secretion. Possibly, the effect on cytokines may be via suppression of NF-κB translocation.

Effects of Topical Corticosteroids and Nonsteroidal Anti-Inflammatory Drugs on Prostaglandin E2-Induced Aqueous Flare Elevation in Pigmented Rabbits

We evaluated the effects of anti-inflammatory potency of corticosteroids and nonsteroidal anti-inflammatory drugs on prostaglandin E2 (PGE2)-induced aqueous flare elevation in pigmented rabbits. Transcorneal diffusion of PGE2, 25 µg/ml (7.09 × 10–2 mmol/l), with the use of a glass cylinder was achieved to produce aqueous flare elevation. Anti-inflammatory drugs were topically administered once before PGE2 application. Aqueous flare was measured with a laser flare-cell meter. Topical single instillation of dexamethasone sodium metasulfobenzoate 0.1%, dexamethasone sodium phosphate 0.1%, and fluorometholone 0.1% 6 h before PGE2 application inhibited 56, 59, and 43% of flare elevation, respectively. Topical single instillation of bromfenac sodium 0.1% and pranoprofen 0.1% 1 h before PGE2 application inhibited 33 and 15% of flare elevation, respectively. Indomethacin 0.5% did not inhibit flare elevation. Corticosteroid eyedrops needed several hours from topical instillation to exhibit inhibition of flare elevation. Most nonsteroidal anti-inflammatory drug eyedrops inhibited aqueous flare elevation when instilled 1 h before PGE2 application.

Effect of Berberine on Interleukin 8 and Monocyte Chemotactic Protein 1 Expression in a Human Retinal Pigment Epithelial Cell Line

Purpose: The aims of this study were to examine the effects of berberine, an alkaloid isolated from some medicinal herbs, on interleukin 8 (IL-8) and monocyte chemotactic protein 1 (MCP-1) expression in a human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin 1β (IL-1β) or tumor necrosis factor α (TNF-α). Methods: ARPE-19 cells were cultured to confluence. Berberine and IL-1β or TNF-α were added to the medium. IL-8 mRNA and MCP-1 mRNA were measured by semiquantitative reverse-transcription polymerase chain reaction and real-time polymerase chain reaction. IL-8 and MCP-1 protein concentrations in the media were measured using enzyme-linked immunosorbent assay. Results: Berberine dose-dependently inhibited IL-8 mRNA and MCP-1 mRNA expression of the cells and protein levels in the media stimulated with IL-1β or TNF-α. Conclusion: These findings indicate that berberine dose-dependently inhibited the expression of IL-8 and MCP-1 induced by IL-1β or TNF-α.

Effects of oral administration of Gardeniae fructus extract and intravenous injection of crocetin on lipopolysaccharide- and prostaglandin E2-induced elevation of aqueous flare in pigmented rabbits

The purpose of this study was to evaluate the effects of extracts of Coptidis rhizoma, Phellodendri cortex and Gardeniae fructus, which are medicinal herbs in Orengedoku-to (Huanglin-Jie-Du-Tang in Chinese), and crocetin (a major component of Gardeniae fructus) on experimental elevation of aqueous flare in pigmented rabbits. To produce aqueous flare elevation, 0.5 microg/kg lipopolysaccharide (LPS) was injected into the ear vein, or prostaglandin E2 (PGE2) 25 microg/ml, was applied to the cornea by means of a glass cylinder. Animals were pretreated by oral administration of 150 g/day of food containing 0.15% (w/w) extract powder of Coptidis rhizoma, 0.10% (w/w) extract powder of Phellodendri cortex or 0.15% (w/w) extract powder of Gardeniae fructus for 4 days, or by intravenous injection of crocetin, 0.3, 3, 30 or 300 microg/kg, 30 minutes before aqueous flare elevation. Aqueous flare was measured with a laser flare-cell meter. Aqueous flare intensity was expressed as the area under the curve (AUC) in arbitrary units. The AUC of LPS- and PGE2-induced aqueous flare elevation was 4685 and 1386 arbitrary units, respectively. Pretreatment by oral administration of 0.15% (w/w) extract of Coptidis rhizoma or 0.10% (w/w) extract of Phellodendri cortex did not inhibit LPS-induced aqueous flare elevation. Pretreatment by oral administration of 0.15% extract of Gardeniae fructus suppressed LPS-induced aqueous flare elevation (AUC: 1411 arbitrary units). Pretreatment by intravenous injection of 3, 30 or 300 microg/kg of crocetin-inhibited LPS-induced aqueous flare elevation in a dose-dependent manner. Pretreatment with 3 or 30 microg/kg of crocetin did not inhibit PGE2-induced aqueous flare elevation, but 300 microg/kg of crocetin inhibited PGE2-induced aqueous flare elevation (AUC: 918 arbitrary units).

Use of a hyperdried cross-linked amniotic membrane as initial therapy for corneal perforations

PurposeTo report the use of hyperdried cross-linked (HDCL) amniotic membrane (AM) patching with tissue adhesive as an initial therapy for corneal perforations.MethodsCryopreserved AM was cross-linked with 0.1% glutaraldehyde and then dried using far-infrared rays and microwaves (hyperdry method). Three eyes of three patients with corneal perforations of up to 3 mm in diameter were included in this study. They were treated with a single-layer patch of HDCL-AM applied with a tissue adhesive (2-octyl-cyanoacrylate). We also evaluated the resistance of HDCL-AM to collagenases during in vitro digestion testing.ResultsIn all three cases, the corneal perforations were repaired within 28 days (range, 17–28 days). No recurrence occurred during the follow-up period (3–6 months). In the collagenase digestion testing, the HDCL-AM did not dissolve until 48 h, whereas the cryopreserved AM completely dissolved within 60 min.ConclusionsThree cases of corneal perforations were successfully managed using HDCL-AM patching with tissue adhesive. The HDCL-AM was resistant to collagenases during in vitro digestion testing. The HDCL-AM was a useful substrate for corneal perforations. This simple surgical technique may be one of the initial therapeutic options for corneal perforations.

Effects of topical instillation of traditional herbal medicines, herbal extracts, and their components on prostaglandin E2-induced aqueous flare elevation in pigmented rabbits.

PURPOSE To evaluate the effect of topical instillation of traditional herbal medicines, herbal extracts, and their components on the elevation of aqueous flare induced by prostaglandin E(2) (PGE(2)) in pigmented rabbits. METHODS Transcorneal diffusion of 25 micro g/mL of PGE(2) was carried out through a glass cylinder placed on the cornea to induce aqueous flare elevation in pigmented rabbits. Traditional herbal medicines, herbal extracts, and their components were topically instilled before the PGE(2) application. Aqueous flare was measured with a laser flare-cell meter. RESULTS Two instillations, 60 and 30 minutes before PGE(2), of Kakkon-to, Sairei-to, Orengedoku-to, Senkanmeimoku-to, Scutellariae radix extract, Coptidis rhizoma extract, Gardeniae fructus extract, Phellodendri cortex extract, baicalein, baicalin, wogonin, crocetin, berberine, or glycyrrhizine did not inhibit the elevation induced by PGE(2). Two instillations, 60 and 30 minutes before PGE(2), of a Ligusticum wallichii extract (100 mg/mL) inhibited the elevation by 20%. Two instillations (5 and 3 hours before PGE(2)) of baicalein (1 mg/mL) or baicalin (5 mg/mL) inhibited the elevation by 16% and 24%, respectively. Two instillations, 5 and 3 hours before PGE(2), of wogonin, crocetin, berberine, or glycyrrhizine did not inhibit the elevation. CONCLUSION Two instillations of Ligusticum wallichii extract 60 and 30 minutes before the PGE(2), and two instillations of baicalein or baicalin, 5 and 3 hours before the PGE(2), inhibited the PGE(2)-induced aqueous flare elevation in pigmented rabbits.

Effect of N-Acetylcysteine on Lipopolysaccharide- Induced Uveitis in Rats

PurposeIn this study we investigated the in vivo effect of N-acetylcysteine (NAC) on lipopolysaccharide (LPS)-induced uveitis in rats.MethodsTo induce uveitis, LPS (100 µg) was injected into subcutaneous tissue of Wistar rats (170–190 g). NAC was injected intraperitoneally. Intracameral levels of protein, cells, nitrite, tumor necrosis factor (TNF)-α, and interleukin (IL)-6 were determined by spectrophotometry, hemocytometry, and enzyme-linked immunosorbent assay. Expression of TNF-α, IL-6, endothelial leukocyte adhesion molecule-1 (E-selectin), intercellular adhesion molecule-1 (ICAM-1), and inducible nitric oxide synthase (iNOS) mRNA was examined by real-time polymerase chain reaction.ResultsLPS injection elevated intracameral protein and cells, and the elevation was inhibited by NAC. LPS injection induced expression of TNF-α, IL-6, E-selectin, and ICAM-1 mRNA in the iris/ciliary body at 3 h, and iNOS mRNA at 6 h. The LPS-induced elevation of the mRNA levels was inhibited by NAC. NAC inhibited LPS-induced intracameral elevation of TNF-α, IL-6, and nitrite.ConclusionNAC decreased LPS-induced uveitis in vivo by reducing the expression of proinflammatory cytokines and adhesion molecules. Jpn J Ophthalmol 2007;51:14–20

Effects of Intravenous Administration of FR122047 (a Selective Cyclooxygenase 1 Inhibitor) and FR188582 (a Selective Cyclooxygenase 2 Inhibitor) on Prostaglandin-E2-Induced Aqueous Flare Elevation in Pigmented Rabbits

Purpose: Two isoforms of cyclooxygenase (COX-1 and COX-2) exist. To determine in vivo effects of the intravenous administration of FR122047 (a selective COX-1 inhibitor), FR188582 (a selective COX-2 inhibitor), diclofenac sodium or dexamethasone phosphate disodium on prostaglandin-E2 (PGE2)-induced aqueous flare elevation and mRNA levels for COX-1 and COX-2 in pigmented rabbits. Methods: To produce aqueous flare elevation in rabbits, PGE2, 25 µg/ml, was applied to the cornea with the use of a glass cylinder. FR122047, FR188582, diclofenac sodium or dexamethasone phosphate disodium was intravenously injected before PGE2 application. Aqueous flare was measured with a laser flare-cell meter. The mRNA levels for COX-1 and COX-2 in the iris-ciliary body were determined by real-time polymerase chain reaction. Results: FR122047, FR188582 and diclofenac sodium (15 µmol/kg each) injected intravenously 30 min before PGE2 application inhibited 29 ± 5, 40 ± 12 and 50 ± 9% of aqueous flare elevation, respectively. Simultaneous injection of FR122047 (15 µmol/kg) and FR188582 (15 µmol/kg) 30 min before PGE2 application inhibited 61 ± 8% of flare elevation. Dexamethasone phosphate disodium (15 µmol/kg) injected intravenously 300 min before PGE2 application inhibited 68 ± 8% of aqueous flare elevation. Less than 3-fold changes in mRNA levels for COX-1 and COX-2 in the iris-ciliary body were noted after PGE2, FR122047, FR188582, diclofenac sodium or dexamethasone phosphate disodium treatment. Conclusion: It is possible that enzyme activities of both COX-1 and COX-2 may be involved in the mechanism of PGE2-induced aqueous flare elevation in pigmented rabbits.