interleukin-8 and Pterygium

Topics: Biomarkers; Early Diagnosis; Humans; Interleukin-6; Interleukin-8; Prognosis; Pterygium; Recurrence; Tears; Vascular Endothelial Growth Factor A

Pterygium is a common eye disease, linked to an increased exposure to UV radiation and dry environments. The associated pathology culminates in visual impairment and, in some rare cases, blindness. However, there remains a lot of uncertainty concerning the pathogenesis of this fibrovascular lesion. As the composition of the tear film provides a reflection into the pathological changes at the ocular surface, tear analysis represents an ideal approach to gain insight in the progression of disease following pterygiectomy. This study enrolled 19 patients and age/gender-matched healthy controls. Tear film levels of interleukin- (IL-) 6, IL-8, and vascular endothelial growth factor (VEGF) were investigated over time, and preoperative concentrations were linked to corneal neovascularization and pterygium size. Diminished tear film levels were found in unilateral patients who show no clinical signs of pterygium recurrence over a period of one year. Hence, our results highlight the potential of using the course of IL-6, IL-8, and VEGF levels in tears as biomarkers for recovery. In addition, when focusing on the affected eyes (i.e., primary and recurrent pterygium), we detected fold changes in preoperative cytokine concentrations to correspond with disease severity. As our proposed biomarkers did not reveal a linear relationship with corneal neovascularization nor the invasive behaviour of pterygium, no exact role in the pterygium pathology could be established. Hence, our data point to these factors being contributors rather than decisive players in the pathological processes.

Topics: Adult; Aged; Case-Control Studies; Cytokines; Female; Humans; Interleukin-6; Interleukin-8; Male; Middle Aged; Prospective Studies; Pterygium; Tears; Vascular Endothelial Growth Factor A

Intraoperative mitomycin C (MMC) is widely used to prevent pterygium recurrence and glaucoma filtering bleb failure, but it has been shown to induce corneal inflammation and cell death. Postoperative dexamethasone (DEX) is advocated to reduce MMC-related inflammation and cell death in corneal fibroblasts. Nevertheless, its long-term regulation mechanism in Tenon's capsule remains to be explored. The purpose of this study was to investigate how DEX modifies MMC's effects in human Tenon's capsule fibroblasts (HTFs). HTFs isolated from the pterygium surgical patients (n = 6) were treated with MMC at 0, 0.1, 0.2, and 0.4 mg/ml for 5 min and incubated in DEX at 10 μM for 0, 1, 2, and 3 days. Recombinant interleukin-8 (IL-8) was used to verify the effect of MMC-related IL-8 secretion. Cell proliferation of all the treated cells was analyzed by WST-1 assay. The amount of IL-8 secretion in HTFs was determined by enzyme-linked immunosorbent assay. Immunoblotting assay was used to analyze the expression of peroxisome-proliferator-activated receptor gamma (PPARγ) and B-cell lymphoma-extra large (Bcl-xL). Our results revealed that MMC significantly reduced the HTF cell proliferation rate. Additionally, MMC significantly upregulated IL-8 secretion in HTFs concentration-dependently. At 3 days post treatment (dpt), 5-min exposures to 0.1, 0.2, and 0.4 mg/ml MMC resulted in 1.4-fold (p = 0.012), 1.6-fold (p = 0.012), and 2.5-fold (p = 0.001) increases of IL-8 secretion. In contrast, DEX reversed the MMC-retarded cell proliferation rate (p = 0.036) and repressed MMC-related IL-8 secretion by 33.5% at 3 dpt (p = 0.003). Addition of recombinant IL-8 noticeably suppressed HTF cell proliferation in a concentration-dependent manner. DEX stimulated upregulation of both PPARγ and Bcl-xL at 1 dpt in normal HTFs and at 2 dpt in MMC-treated HTFs. PPARγ silencing reduced expression of PPARγ and Bcl-xL, but enhanced IL-8 secretion (p < 0.001). On the other hand, Bcl-xL silencing enhanced IL-8 secretion (p < 0.001), but did not affect PPARγ expression. These revealed that IL-8 secretion in HTFs is modulated by PPARγ-dependent Bcl-xL signaling. We conclude that DEX reversed the MMC-inhibited HTF cell proliferation via diminishing the MMC-induced IL-8 secretion, which resulted from a late-phase upregulation of the PPARγ and Bcl-xL. These long-term effects suggest a beneficial postoperative DEX treatment following intraoperative MMC application.

Topics: Apoptosis; bcl-X Protein; Blotting, Western; Cell Proliferation; Cells, Cultured; Dexamethasone; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gene Expression Regulation; Glucocorticoids; Humans; Interleukin-8; Mitomycin; PPAR gamma; Pterygium; RNA; Tenon Capsule

Pterygium is a proliferative, inflammatory, and invasive ocular surface disease associated with excessive ultraviolet (UV) exposure. This investigation was conducted to identify UV activated signaling pathways in pterygium epithelial cells (PECs) that mediate cytokine and growth factor production and to determine whether these pathways are sensitive to blockade by anti-inflammatory agents such as retinoic acid (RA) and interferon (IFN)-alpha.. PECs were pretreated with or without inhibitors of the ERK1/2, JNK, and p38 (PD98059, SB202190, and SB203580, respectively) mitogen-activated protein kinases (MAPK) or with inhibitors of the tyrosine kinase activity of epidermal growth factor receptor (EGFR; PD153035) and platelet-derived growth factor (PDGF; AG1295); exposed to UVB (20 mJ/cm2); and then further treated with the same inhibitors. Media were harvested and analyzed by ELISA for interleukin (IL)-6, IL-8, and vascular endothelial growth factor (VEGF). Cytokine mRNA was assessed by reverse transcription-polymerase chain reaction (RT-PCR).. Inhibitors of ERK1/2, JNK, and p38 MAPKs significantly abolished the UVB-mediated increase in IL-6, IL-8, and VEGF. PD153035 reduced IL-8, AG1295 repressed IL-6, and both inhibitors partially downregulated VEGF production in UV-exposed PECs. RA and IFN-alpha dose dependently abrogated IL-6 and IL-8 but had no effect on VEGF expression after UV exposure.. The results have identified a stress-induced intracellular pathway and potential cell-surface transmitters that may be relevant to pterygium development. Moreover, two anti-inflammatory/antiangiogenic agents were identified that reduced cytokine production in the study model. Topical application of these drugs may benefit patients with pterygia, potentially reducing the necessity for surgical intervention.

Topics: Cells, Cultured; Cytokines; Dose-Response Relationship, Drug; Enzyme Inhibitors; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Growth Substances; Humans; Interferon-alpha; Interleukin-6; Interleukin-8; MAP Kinase Signaling System; Pterygium; Receptors, Cell Surface; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tretinoin; Ultraviolet Rays; Vascular Endothelial Growth Factor A

Pterygia are common ocular surface lesions that are thought to be induced by exposure to ultraviolet (UV) radiation. The hypothesis tested in the current study is that UV radiation modulates the expression of interleukin (IL)-6 and -8, which could promote the neovascularization and chronic inflammation regularly observed in pterygia.. Immunohistochemical analysis was performed on 10 pterygia and 14 specimens of normal conjunctiva (4 of which contained limbus), to identify the cellular source of these cytokines. Pterygium epithelial cells were exposed to UVB (0-100 mJ/cm(2)) and the expression of cytokine mRNA and protein was determined by reverse transcription-polymerase chain reaction (RT-PCR), RNase protection assay (RPA), and enzyme immunoassay. Similarly, pterygium tissue in organ culture was UVB irradiated and the supernatants analyzed for cytokine production.. IL-6 and -8 proteins were abundantly expressed, predominantly by the pterygium epithelium, with additional IL-8 immunoreactivity associated with the vascular endothelium. In contrast, significantly less staining for both cytokines was observed in normal conjunctiva, cornea, and limbus. Expression of both IL-6 and -8 mRNA and protein was induced in UVB-irradiated pterygium epithelial cells in a time- and dose-dependent manner. Similarly, IL-6 and -8 proteins were significantly elevated in UVB-treated compared with nonirradiated pterygia.. This study provides the first direct experimental evidence that implicates UV in the pathogenesis of pterygia. The two proinflammatory cytokines that are induced by UV radiation may play a key role in the development of pterygia, by initiating blood vessel formation, cellular proliferation, tissue invasion, and inflammation. Strategies aimed at reducing ocular exposure to UV light may decrease the incidence and recurrence of pterygia.

Topics: Cells, Cultured; Conjunctiva; Epithelial Cells; Humans; Immunoenzyme Techniques; Interleukin-6; Interleukin-8; Organ Culture Techniques; Pterygium; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Time Factors; Ultraviolet Rays