Interleukin-6(IL-6)Influences Fibronectin Expression In Cultured Bovine Trabecular Meshwork Cells

BackgroundPrimary open angle glaucoma(PO AG)is a kind of glaucoma that is characterized by open anterior chamber angle and slow elevation of intraocular pressure,it can destory optic nerve and there is no clear pathogenesis about it.Among the known risk factors,the accumulation of extracellular matrix outside the trabecular meshwork is the main cause of this disease.At present,there are many reports that the content of interleukin-6(IL-6)in aqueous humor of POAG patients is lower than that in normal people,but there is no report on the effect of IL-6 on POAG.PurposeTo investigate the changes of fibronectin(FN)expression in cultured bovine trabecular meshwork cells when cells were stimulated by different concentrations of IL-6 to provide evidence for early diagnosis and new therapy of POAG.MethodsWe use Tissue Mass Culture Method to get trabecular meshwork tissue from fresh bovine eyes,then we extract and culture the third generation of bovine trabecular meshwork cells from them.Cell morphology is used to identify the cells.The third generation of bovine trabecular meshwork cells were transplanted on the 6-well cell culture plate.After 24 hours of starvation in serum-free medium,IL-6 was added in different final concentration(0 ng/ml 0.1 ng/ml 0.5 ng/ml 1 ng/ml)for 24 h.Real-time PCR was used to detect the relative expression of FN mRNA in bovine trabecular meshwork cells and western blot was used to detect the relative expression of FN protein.ResultsThe cultured bovine trabecular meshwork cells were fusiform or polygonal.A few pigmented particles were found in the cells under inverted biomicroscopy,which were in accordance with the morphological characteristics of the third generation of bovine trabecular meshwork cells recorded in the literature.Real-time PCR showed that FN mRNA of four groups,bovine trabecular meshwork cells cultured in 0.1 ng/ml,0.5 ng/ml and 1 ng/ml IL-6,was 1.000±0.000,0.213±0.004,0.056±0.001,0.019±0.002 respectively,indicating down-regulated.There was significant difference between each group(0ng/ml,0.1 ng/ml,0.5 ng/ml,1 ng/ml)(P<0.05,Figure 2).In consistently,the expressions of FN protein between every two groups are different(P<0.05,Figure 3,Figure 4),which is1.167±0.012、0.662±0.009、0.238±0.011、0.061±0.011 respectively.ConclusionsCultured bovine trabecular meshwork cells in vitro produced FNs after being stimulated by exogenous IL-6,and the production was negatively correlated with the concentration.We speculate that IL-6 participated in the pathogenesis of POAG and affected its progression.