目的 应用Luminex液相芯片检测原发性开角型青光眼(POAG)患者房水中多种细胞因子浓度,并分析这些因子的改变与疾病的关系.设计实验研究.研究对象24例POAG患者及同年龄段22例老年性白内障患者的房水样本.方法 使用Luminex液相芯片检测房水中10种细胞因子浓度,比较POAG和白内障患者房水中细胞因子浓度的差异.主要指标 血小板衍生生长因子-AA(PDGF-AA)、血小板衍生生长因子-AB(PDGF-AB)、表皮生长因子(EGF)、血管内皮生长因子(VEGF)、中性粒细胞集落刺激因子(G-CSF)、白介素-6(IL-6)、白介素-8(IL-8),趋化因子配体5(RANTES)、干扰素-γ(IFN-γ)和肿瘤坏死因子-α(TNF-α)的浓度.结果 POAG患者房水中PDGF-AB (P=0.040)、EGF(P=0.048)、VEGF(P=0.037)、G-CSF(P=0.029)和IL-6(P=0.018)的水平显著低于老年性白内障患者;PDGF-AA浓度显著高于白内障患者(P=0.001);IL-8、RANTEs、IFN-γ和TNF-α浓度两组间无显著差异(P均＞0.05).POAG患者房水中IL-6(P＜0.001,ρ=-0.778)、IL-8(P=0.001,ρ=-0.623)和TNF-α(P＜0.001,ρ=-0.685)的浓度与视野平均缺损值(MD)显著相关.结论 POAG患者房水中多种细胞因子浓度低于同年龄段老年性白内障患者,其能否反映视神经和小梁网的损害情况有待进一步研究.

Objective To measure cytokine concentrations in aqueous humor of patients with primary open angle glaucoma (POAG) and cataract.Methods A cross-sectional study was performed under the informed consent of each subject from June to December in 2016 in Daping Hospital.A total of 24 cases of POAG patients served as the experimental group,while 22 cases of age related cataract patients served as control group.Concentrations of basic fibroblast growth factor (bFGF),monocyte chemoattractant protein-1 (MCP-1),brain derived neurotrophic factor (BDNF),soluble intercellular adhesion molecule-1 (sICAM-1),soluble neural ceil adhesion molecule (sNCAM),and soluble vascular cell adhesion molecule-1 (sVCAM-1) with Luminex 200 were detected between the two groups.Then the comparison and the correlation analysis were performed between the two groups.Results Concentrations of MCP-1,sVCAM-1 and sNCAM in the experimental group were higher than those in the control group,with significant differences between them (P =O.017,0.007,0.001).The concentration of bFGF-2 in the experimental group was significantly lower than that in the control group (P =0.027).There was no significant difference in the expression levels of BDNF and sICAM-1 between the two groups (P＞0.05).The expression levels of MCP-1,sICAM-1,sNCAM and sVCAM-1 were significantly associated with each other (all at P ＜ 0.05).Conclusions The expression levels of MCP-1,sVCAM-1 and sNCAM are elevated in POAG patients.POAG may be a neurodegenerative disease associated with neuroinflammation.

Background Researches showed that transforming growth factor-β2 (TGF-β2) promotes the activity of human Tenon capsular fibroblasts (TFs),which plays a role in the scarring of filtering blebs after antiglaucoma surgery.However,its mechanism is not fully clear.Lysyl oxidases (LOXs) are important extracellular matrix proteases which can catalyze the cross-linking of collagen and elastin.Investigating the impact of TGF-β2 on the expression of LOXs has a great significance for the understanding of the pathogenesis of filtering bleb scarring and its prevention.Objective This study was to investigate the effect of TGF-β2 on the expression of LOXs in cultured human TFs.Methods The TFs at 4-8 generations were divided into normal control group and different concentrations of TGF-β2 treated-groups,and 100,200,400,800 μ1 of TGF-β2 with the final concentration of 2,4,8 and 16 ng/ml was added into the medium to treat human TFs respectively for 24 hours.The LOXs in the cells were detected by Western blot to determine the optimal dose of TGF-β2.The 4 ng/ml TGF-β2(200 μ1) was used to treat human TFs for 6,12,24 and 48 hours respectively,and the change of LOXs expression in the cells over time was assayed by Western blot.The expression and distribution of LOX protein in the normal cells and TGF-β2-treated cells was detected by using immunofluorescence technique.This study was approved by Daping Hospital of Third Military Medical University Ethic Commission.The guardians of the patients who offered the specimen knew the purpose of the study and signed informed consent.Results Western blot assay showed that the expressions of LOX,LOXL1,LOXL2,LOXL3 and LOXL4 in the cells were gradually elevated from the normal control group and 2,4,8,16 ng/ml TGF-β2-treated groups,showing significant differences among the groups (F =37.338,13.438,31.067,11.767,15.167,all at P＜0.01).The expression of LOXL2 protein in the cells was 0.68±0.07,1.09±0.10,1.32±0.07,1.50± 0.06 and 1.89±0.12 in the normal control group and 6-,12-,24-and 48-hour groups respectively after 4 ng/ml TGF-β2 treatment,with a significant increase over time (F =82.832,P=0.000).The expression of LOX was weak in the normal cultured TFs,while the fluorescence intensity of LOX expression was evidently enhanced in the cytoplasm of the cells in the TGF-β2-treated group.Conclusions TGF-β2 upregulates the expressions of LOXs in human TFs in a dose-and time-dependent manner,which probably offers a basis for the further study on the prevention of filtering bleb scarring after glaucoma surgery.

Objective To compare the difference of the concentration of inflammatory cytokines in the aqueous humor of patients with primary angle-closure glaucoma.Design Experimental study.Participants This study included 16 eyes with acute primary angle-closure glaucoma,16 eyes with chronic primary angle-closure glaucoma,and 22 eyes with cataract.Methods The concentrations of nine cytokines in aqueous humor were measured with the Luminex suspension array technique.The clinical characteristics of patients were also obtained for correlation analysis.Significant level was P=0.0167.Main Outcome Measures The concentrations of granulocyte colony-stimulating factor (G-CSF),interferon-γ (IFN-γ),interleukin-6,(IL-6),interleukin-8 (IL-8),monocyte chemotactic protein-1 (MCP-1),platelet derived growth factor-AA (PDGF-AA),PDGF-AB,RANTES and tumor necrosis factor-α (TNF-α) in aqueous humor.Results Compared with those from the senile cataract group,the concentrations of G-CSF,PDGF-AA,IL-6,IL-8,MCP-1,RANTES and TNF-α were significantly higher in the aqueous humor samples from acute primary angle-closure glaucoma (all P＜0.0167),and the concentrations of G-CSF,PDGF-AA,IL-6,IL-8,and MCP-1 were significantly elevated in the aqueous humor of patients with chronic primary angle-closure glaucoma (all P＜0.0167).There was no significant difference in the concentration of inflammatory factors between acute primary angle-closure glaucoma and chronic primary angle-closure glaucoma (all P＞0.0167).Conclusions In addition to acute angle closure glaucoma,chronic angle closure glaucoma also had inflammation in ocular anterior segment.