I-Mo Fang

Chitosan Oligosaccharides Attenuate Ocular Inflammation in Rats with Experimental Autoimmune Anterior Uveitis

We investigated the protective effects and mechanisms of chitosan oligosaccharides (COS) on experimental autoimmune anterior uveitis (EAAU) in rats. EAAU was induced in Lewis rats by footpad and intraperitoneal injections of melanin-associated antigen. The rats received intraperitoneal injections of low-dose (5 mg/kg) or high-dose (10 mg/kg) COS or PBS daily after the immunization. The effects of COS were evaluated by determining the clinical scores and the morphology of the iris/ciliary body (ICB). The expression of inflammatory mediators was evaluated using western blot, immunofluorescence, and ELISA. Treatment with COS significantly attenuated the clinical scores and the leukocyte infiltration in the ICB in a dose-dependent manner. COS effectively reduced the expression of inflammatory mediators (TNF-α, iNOS, MCP-1, RANTES, fractalkine, and ICAM-1). Moreover, COS decreased the IκB degradation and p65 presence in the ICB, which resulted in the inhibition of NF-κB/DNA binding activity. In an in vitro study, sensitized spleen-derived lymphocytes of the COS-treated group showed less chemotaxis toward their aqueous humor and decreased secretion of the above inflammatory mediators in the culture media. COS treated EAAU by inhibiting the activation of NF-κB and reducing the expression of inflammatory mediators. COS might be a potential treatment for acute anterior uveitis.

Comparative effects of fatty acids on proinflammatory gene cyclooxygenase 2 and inducible nitric oxide synthase expression in retinal pigment epithelial cells.

Dietary fat modification is a promising approach to prevent age-related macular degeneration (AMD). However, which types of fatty acids carry a greater risk for AMD remains unclear. In this study, we compared the effects of 18-carbon fatty acids with different degrees of unsaturation on the expression of the proinflammatory genes cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) in human retinal pigment epithelium (RPE). Additionally, we investigated whether lutein could modulate these genes induced by fatty acids in RPE. Treatment with oleic acid, linoleic acid (LA), or linolenic acid increased the expression of iNOS and COX-2 genes and the production of prostaglandin E(2 )and nitric oxide (NO) in RPE, whereas the saturated fatty acid stearic acid had little effect on these genes. Of the fatty acids studied, LA had the greatest effects on the induction of these genes. Furthermore, LA also induced NF-kappaB transcriptional activation the most. Lutein inhibited LA-induced expression of COX-2 and iNOS in a dose-dependent manner. These data suggested that specific unsaturated fatty acids, particularly LA, can stimulate RPE cells to express proinflammatory genes, which may contribute to the pathogenesis of AMD. Lutein inhibited the expression of these genes induced by LA through blockade of NF-kappaB activation.

Docosahexaenoic acid reduces linoleic acid induced monocyte chemoattractant protein‐1 expression via PPARγ and nuclear factor‐κB pathway in retinal pigment epithelial cells

SCOPE To investigate whether docosahexaenoic acid (DHA) could inhibit linoleic acid (LA) induced monocyte chemoattractant protein (MCP)-1 expression in human retinal pigment epithelial (RPE) cells. METHODS AND RESULTS ARPE-19 cells were pretreated with DHA and then exposed to LA. The expression of MCP-1 and PPARγ was examined using RT-PCR and Western blot analysis. LA at 10, 25, or 50 μM induced expression of MCP ARPE-19 cells in a dose-dependent manner (p < 0.05). DHA at 50 and 100 μM effectively inhibited LA-induced MCP-1 expression and production (p < 0.05) and NF-κB activation. In addition, the culture medium from LA-stimulated ARPE-19 cells could induce tube formation in choroidal endothelial cells (RF6A), whereas 100 μM DHA inhibited tube formation. DHA at 100 μM increased the expression and activity of PPARγ (p < 0.05). Pretreatment with PPARγ inhibitor (GW9662) abolished the inhibitory effect of DHA (100 μM) on LA-induced IκB degradation, p65 translocation, and MCP-1 expression in ARPE-19 cells (p < 0.05), as well as tube formation in RF6A. CONCLUSION DHA reduced LA-induced MCP-1 expression via a PPARγ- and NF-κB-dependent pathway in ARPE-19 cells. These results suggest the molecular mechanisms underlying the beneficial effects of increased consumption of DHA and reduced consumption of LA on age-related macular degeneration.

Induced Pluripotent Stem Cells Without c-Myc Ameliorate Retinal Oxidative Damage via Paracrine Effects and Reduced Oxidative Stress in Rats

PURPOSE To investigate the efficacy and mechanisms of non-c-Myc induced pluripotent stem cell (iPSC) transplantation in a rat model of retinal oxidative damage. METHODS Paraquat was intravitreously injected into Sprague-Dawley rats. After non-c-Myc iPSC transplantation, retinal function was evaluated by electroretinograms (ERGs). The generation of reactive oxygen species (ROS) was determined by lucigenin- and luminol-enhanced chemiluminescence. The expression of brain-derived neurotrophic factor, ciliary neurotrophic factor, basic fibroblast growth factor (bFGF), stromal cell-derived factor (SDF)-1α, and CXCR4 was measured by immunohistochemistry and ELISA. An in vitro study using SH-SY5Y cells was performed to verify the protective effects of SDF-1α. RESULTS Transplantation of non-c-Myc iPSCs effectively promoted the recovery of the b-wave ratio in ERGs and significantly ameliorated retinal damage. Non-c-Myc iPSC transplantation decreased ROS production and increased the activities of superoxide dismutase and catalase, thereby reducing retinal oxidative damage and apoptotic cells. Moreover, non-c-Myc iPSC transplantation resulted in significant upregulation of SDF-1α, followed by bFGF, accompanied by a significant improvement in the ERG. In vitro studies confirmed that treatment with SDF-1α significantly reduced apoptosis in a dose-dependent manner in SH-SY5Y cells. Most transplanted cells remained in the subretinal space, with spare cells expressing neurofilament M markers at day 28. Six months after transplantation, no tumor formation was seen in animals with non-c-Myc iPSC grafts. CONCLUSIONS We demonstrated the potential benefits of non-c-Myc iPSC transplantation for treating oxidative-damage-induced retinal diseases. SDF-1α and bFGF play important roles in facilitating the amelioration of retinal oxidative damage after non-c-Myc iPSC transplantation.

Expression of RANTES and CCR5 in Rats with Experimental Autoimmune Anterior Uveitis

Objective: To determine the pattern of expression of RANTES and its receptor, CCR5, and to establish their roles in experimental autoimmune anterior uveitis, an animal model of human acute anterior uveitis. Methods: Uveitis was induced in the Lewis rats with the injection melanin-associated antigen into the left footpad. The rats were scored for the development of clinical disease. At defined tine points, RANTES and CCR5 mRNA expression were semiquantified by using RT-PCR. The concentration of RANTES protein in aqueous humor was quantified by ELSIA. Results: RANTES mRNA started increasing on day 11, concurrent with the infiltration of leukocytes in aqueous humor and onset of clinical disease. The concurrent with the infiltration of leukocytes in aqueous humor and onset of clinical disease. The concentration of RANTES protein also increased on day 11, Furthermore in accord with its ligand RANTES contributed to the recruitment of inflammatory cells into the eye and to the disease activity. RANTES could be potential targets for acute anterior uveitis therapy.