国产环孢素A眼液治疗大鼠角膜移植术后免疫排斥反应的研究

摘要

目的　观察环孢素A(cyclosporine, CsA)滴眼液对鼠角膜移植术后免疫排斥反应的治疗效果。方法　随机对纯系大鼠LOU 65只为受体及F344纯系大鼠33只为供体建立的角膜移植术后免疫排斥反应动物模型分为6组，各组分别于手术后滴用0.5%CsA、1.0%CsA、2.0%CsA、糖皮质激素、糖皮质激素和1.0%CsA及CsA基质滴眼液，观察角膜移植术后植片排斥反应指数(rejection index, RI)，植片平均存活时间(means survival time, MST)，并对角膜植片进行病理学检查及免疫病理学分析。结果 0.5%CsA、1.0%CsA、2.0%CsA、糖皮质激素及混合组的植片MST均较对照组延长1.63～8.46 d，其中2.0%CsA组、糖皮质激素组及混合组的植片MST延长与其他组比较差异有显著性（t≥2.28，P<0.01），混合组为（15.71±5.06） d。对照组的角膜植片排斥RI较其他组高，其中2.0%CsA组与糖皮质激素组的角膜植片排斥RI一致，分别为5.34±1.92和5.18±1.69；混合组治疗效果最好，RI为4.31±1.50，而CsA滴眼液治疗后大鼠血清中的浓度介于21.6～62.2 mg/L。病理学检查：各治疗组角膜植片淋巴细胞浸润减轻，新生血管减少；免疫病理显示，除0.5%CsA组外，其余各组的淋巴细胞相关抗原1（LFA-1，CD+11a）、ICAM-1表达和巨噬细胞浸润均明显减少。结论　环孢素A滴眼液能够抑制大鼠角膜移植术后免疫排斥反应的发生，与糖皮质激素联合应用能取得更佳的效果。%Objective　To evaluate the efficacy of topical cyclosporine A (CsA) on a penetrating keratoplasty rejection model in rats. Methods　Lou rats received orthotopic corneal allografts from inbred F344 donors. The rats were treated with one drop of topical solution four times daily for 30 days. Sixty rats were divided into one control (the matrix of CsA solution) and five treatment groups (0.5%, 1.0%, 2.0% CsA, glucocorticoid, glucocorticoid and 1.0% CsA). Rejection index (RI) and mean survival time (MST) were calculated, and histopathological and immunohistopathological examination were performed for each group. Results The graft MST was postponed by topical CsA treatment. There was significant difference between each treatment group and the control group (t≥2.28, P<0.0l), especially 2% CsA, steroid and combined 1%CsA and steroid groups, the combined group being the most effective. The RI was lower in all the groups with topical CsA than that in the control group (P<0.01). The effect of 2% CsA was the same as that of the steroid, while 1% CsA combined steroid was the most effective. The histopathological findings confirmed that the topical application of CsA reduced the inflammatory cells infiltrating the graft stroma and the neovascularization. Immunohistopathological analysis demonstrated that there were fewer CD+11a T cells and macrophages in the infiltration and intercellular adhesion molecule (ICAM)-1 positive cells in keratocytes and endothelium of grafts in the treatment groups than in the control group. Conclusions　Topical CsA can inhibit the keratoplasty rejection episodes in the rat and afford better results when combined with topical steroid.