ABSTRACT
PURPOSE: To determine whether insulin-like growth factor (IGF-1) affects transforming growth factor (TGF-beta)- mediated fibronectin accumulation in human lens epithelial cell line (HLE B-3) cells. MATERIALS AND METHODS: HLE B-3 cells were incubated for 24 hours with TGF-beta (10ng/ mL), IGF-1 (10ng/mL), or both. Expression of the fibronectin gene was determined using a real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Fibronectin levels were examined using western blot analysis and immunofluorescence staining. RESULTS: Expression of the fibronectin gene was not different between the TGF-beta/IGF-1 treated group and the TGF-beta treated group (p= 0.116). However, western blot analysis demonstrated decreased fibronectin levels in human lens epithelial cells treated with TGF-beta and IGF-1 compared to those treated with TGF-beta only (p < 0.01). Immunofluorescence staining disclosed inhibition of TGF-beta-induced fibronectin in the presence of IGF-1. CONCLUSION: This study suggests that IGF-1 counteracts TGF-beta-mediated fibronectin accumulation in human lens epithelial cells.
Subject(s)
Humans , Cell Line , Epithelial Cells/cytology , Fibronectins/genetics , Fluorescent Antibody Technique , Gene Expression Regulation/drug effects , Insulin-Like Growth Factor I/pharmacology , Lens, Crystalline/cytology , Reverse Transcriptase Polymerase Chain Reaction , Transforming Growth Factor beta/pharmacologyABSTRACT
PURPOSE: We collected tear or scleral tissues of necrotizing scleritis after pterygium excision, and evaluated them for tumor necrosis factor (TNF)-alpha and matrix metalloproteinase (MMP)-9 to elucidate the molecular basis and seek for treatment of this disease. METHODS: Three patients with necrotizing scleritis after pterygium excision were evaluated for MMP-9 and TNF-alpha in tear and scleral tissue by Western blot analysis. RESULTS: Before treatment with corticosteroid, the patients' tear samples showed increased expression of TNF-alpha and MMP-9 compared to those of the contralateral eye. After treatment, the expression of TNF-alpha and MMP-9 was decreased compared to those of the pre-treated tear samples. The patients' sclera showed increased expression of MMP-9 compared to that of the donors' sclera and the patients' conjunctiva. CONCLUSIONS: Our results suggest that cytokine-related inflammation plays a role in the pathophysiology of necrotizing scleritis and strongly supports, under the guarantee of negative microbiological culture, the prompt use of corticosteroid and immunosuppressive agents to help suppress the progression of this disease.
Subject(s)
Humans , Blotting, Western , Conjunctiva , Immunosuppressive Agents , Inflammation , Matrix Metalloproteinase 9 , Pterygium , Sclera , Scleritis , Tears , Tumor Necrosis Factor-alphaABSTRACT
PURPOSE: We evaluated the efficacy of the PCR which is known as more sensitive method than culture in the diagnosis of causal microorganisms of the infective endophthalmitis. METHODS: We used 0.3 ml of aqueous humor and 0.5 ml of vitreous sample in 3 cases of postoperative and 1 case of endogenous endophthalmitis for detecting causal microorganisms. To detect the bacteria we used universal, Gram positive and negative primers, and to detect the fungus we used fungal primer. RESULTS: Three cases of endophthalmitis, there was no bacteria in the bacterial culture for 10 days but PCR results identified causal microorganisms in all cases. CONCLUSIONS: PCR is effective in the fast and accurate diagnosis of infective endophthalmitis and especially in the culture-negative endophthalmitis.