来氟米特改善 Lewis 大鼠实验性自身免疫性葡萄膜炎。
Amelioration of experimental autoimmune uveitis by leflunomide in Lewis rats.
机构信息
School of Pharmacy, Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Medical University, Hefei, China.
出版信息
PLoS One. 2013 Apr 23;8(4):e62071. doi: 10.1371/journal.pone.0062071. Print 2013.
PURPOSE
To investigate the efficacy of leflunomide in experimental autoimmune uveitis (EAU) in rats.
METHODS
Lewis rats were immunized with interphotoreceptor retinoid-binding peptide (IRBP) in order to generate EAU. Rats received three dose of leflunomide through intragastric administration (prevention or treatment protocols) after immunization at three separate doses (3 mg/kg/d; 6 mg/kg/d; 12 mg/kg/d). Cyclosporin A was administered as a positive) control. Rats were euthanized during peak disease activity (day 14 or 15). Treatment effectiveness was evaluated in vivo using clinical EAU scoring (d14) and histopathological evaluation of enucleated eyes after experimental termination. The expression levels of inflammatory cytokines in the serum were quantified by ELISA. Eyeball of rats were harvested and mRNA expression of interleukin 17 (IL17) and IFN-γ were quantified through RT-PCR. Intracellular expression of interleukin (IL)-17 in the activated CD4(+) T cells was assessed by flow cytometry. The effects of leflunomide inhibition on immune responses in rats were investigated in isolated lymphocytes.
RESULTS
Histopathological and clinical data revealed severe intraocular inflammation in the immunized rat. Inflammation reached its peak on day 14 in this EAU model. Treatment with leflunomide significantly prevented and treated EAU-induced ocular inflammation and decreased clinical and pathological scores compared to vehicle-treated eyes. Gene expression of IL17 and IFN-γ was markedly reduced in leflunomide-treated eyes. Leflunomide significantly decreased the serum levels of IL17 and IFN-γ. The study of IL17+ T cells in peripheral blood and spleen by flow cytometry showed a decreased number of Th17 cell in rats of leflunomide prevented group. Lymphocytes from animals treated with leflunomide had decreased antigen-specific proliferation in vitro compared with lymphocytes from untreated animals.
CONCLUSIONS
Oral administration of leflunomide effectively suppressed IRBP-induced uveitis in rats. These results suggest that leflunomide may be potentially clinical application in uveitis.
目的
研究来氟米特（leflunomide）在大鼠实验性自身免疫性葡萄膜炎（EAU）中的疗效。
方法
用 interphotoreceptor retinoid-binding peptide（IRBP）免疫 Lewis 大鼠，以产生 EAU。大鼠在免疫后分 3 次给予来氟米特（3 mg/kg/d；6 mg/kg/d；12 mg/kg/d），通过灌胃给药。环孢素 A 作为阳性对照。在疾病活动高峰期（第 14 或 15 天）处死大鼠。通过临床 EAU 评分（第 14 天）和实验结束时眼球摘除法的组织病理学评估，在体内评估治疗效果。通过 ELISA 定量检测血清中炎性细胞因子的表达水平。采集大鼠眼球，通过 RT-PCR 定量检测白细胞介素 17（IL17）和 IFN-γ 的 mRNA 表达。通过流式细胞术评估激活的 CD4(+) T 细胞中白细胞介素（IL）-17 的细胞内表达。通过分离的淋巴细胞研究来氟米特抑制对大鼠免疫反应的影响。
结果
组织病理学和临床数据显示，免疫大鼠眼内有严重的炎症。在该 EAU 模型中，炎症在第 14 天达到高峰。与 vehicle 治疗的眼睛相比，来氟米特治疗显著预防和治疗了 EAU 诱导的眼部炎症，并降低了临床和病理评分。来氟米特治疗眼的 IL17 和 IFN-γ 基因表达明显降低。来氟米特显著降低了血清中 IL17 和 IFN-γ 的水平。通过流式细胞术研究外周血和脾脏中的 IL17+T 细胞，发现来氟米特预防组大鼠 Th17 细胞数量减少。与未治疗动物的淋巴细胞相比，来自来氟米特治疗动物的淋巴细胞体外抗原特异性增殖减少。
结论
口服来氟米特可有效抑制大鼠 IRBP 诱导的葡萄膜炎。这些结果表明，来氟米特可能具有在葡萄膜炎中的潜在临床应用价值。
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