环孢素A纳米粒滴眼液抑制大鼠高危角膜移植免疫排斥反应的实验研究

眼科 ›› 2012, Vol. 21 ›› Issue (2): 116-120.

环孢素A纳米粒滴眼液抑制大鼠高危角膜移植免疫排斥反应的实验研究

张海娟, 马科, 游玉霞, 吴雁, 徐清

100005首都医科大学附属北京同仁医院北京同仁眼科中心北京市眼科研究所（张海娟马科、游玉霞）；100190国家纳米技术研究中心（吴雁、徐清）

收稿日期:2011-12-08 出版日期:2012-03-25 发布日期:2012-04-05
通讯作者: 马科，Email：cdmake@sohu.com
基金资助:
北京市科技计划课题资助项目（Z09050700620908）

Cyclosporine A nano-particle eye drops prevent the high risk corneal transplantation immune rejection in rats

ZHANG Hai-Juan, MA Ke, YOU Yu-Xia, WU Yan, XU Qing

Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, 100005 Beijing, China. WU Yan, XU Qing. National ninao-meter research center, 10190 Beijing, China.

Received:2011-12-08 Online:2012-03-25 Published:2012-04-05
Contact: MA Ke, Email: cdmake@sohu.com

摘要/Abstract

摘要： 目的探讨环孢素A纳米粒滴眼液对大鼠高危角膜移植术后排斥反应的抑制作用。设计实验研究。研究对象 Lewis大鼠50只为受体，F344大鼠25只为供体。方法 50只Lewis大鼠右眼采用缝线法诱导角膜新生血管形成，第7天新生血管到达角膜中周部时，全部行穿透性角膜移植术。因术后滴眼液不同将大鼠随机分为5组（A到E组，每组10只大鼠），A组为对照组，给予环孢素A纳米粒滴眼液的基质点药，B组为普通环孢素A滴眼液组，C组、D组、E组分别为0.5%、1%、2%环孢素A纳米粒滴眼液组。术后各组滴用相应的滴眼液，每日3次。拍摄大鼠角膜照片每日1次至14天，以后每2日拍摄角膜照片1次至28天。以角膜植片水肿、混浊、新生血管评分之和为角膜排斥反应指数（RI），记录角膜存活时间（从角膜移植开始到RI=6的时间），术后第14天每组取2只大鼠眼球，以光镜进行角膜病理学观察。主要指标 角膜植片存活时间、角膜排斥反应指数（RI），角膜的病理学改变。结果 A到E组角膜存活时间分别为（7.20±2.93）天、（10.89±3.62）天、（10.50±3.40）天、（11.13±3.94）天和（12.80±4.07）天，各组间比较差异有统计学意义（F=3.20，P=0.022）。B、C、D、E组与A组相比，角膜存活时间明显延长，差异均有统计学意义（P=0.031、0.047、0.027、0.001）。B组与C、D、E组之间比较差异均无统计学意义（P=0.815、0.853、0.255），C组与D、E组之间差异均无统计学意义（P=0.716、0.161），D组与E组之间差异无统计学意义（P=0.333）。A到E组RI分别为7.80±1.48、5.67±1.80、6.10±1.66、5.25±1.75和4.80±1.23，组间比较差异具有统计学意义（F=5.202，P=0.002）。B、C、D、E组与A组相比，RI明显减小，差异有统计学意义（P=0.005、0.021、0.002、0.000）。B组与C、D、E组比较差异均无统计学意义（P=0.555、0.592、0.241）。C组与D、E组比较差异无统计学意义（P=0.265、0.074）。D组与E组之间差异无统计学意义（P=0.553）。A组角膜植片明显水肿、增厚，基质层可见大量细胞浸润，细胞排列紊乱。B~E组角膜基质层浸润程度较A组明显减轻。结论普通环孢素滴眼液组及高、中、低浓度的环孢素A纳米粒滴眼液均能有效地抑制高危角膜移植术后的排斥反应。（眼科，2012, 21: 116-120）

关键词: 角膜移植, 免疫排斥, 环孢素A, 纳米粒滴眼液

Abstract: Objective To investigate the preventing effect of cyclosporine A nano-particle eye drops on immune rejection after high risk corneal transplantation. Design Experimental study. Participants Fifty Lewis rats as receptor and twenty-five F344 rats as donator. Method Corneal neoangiogenesis was induced by suture method. Rats were divided into 5 groups(group A to E). All the animals underwent corneal penetrating transplantation in right eyes at 7th day after suturing when the new vascular appeared in the middle part of the cornea. The cornea donator were F344 rats. Group A received vehicle eye drops, group B received cyclosporine A eye drops, group C to E received 0.5%, 1% and 2% cyclosporine A nano-particle eye drops respectively. All animals were received the eye drops 3 times a day. Cornea was observed by slit lamp everyday within 14 days and photographs were taken. Then the cornea was observed every two days until 28 days. The cornea was studied pathologically at day 14 for 2 rats each group. Main Outcome Measures Cornea survival time (from corner transplantation to the time when RI equals 6), rejection index (RI), pathologically observation of cornea at day 14. Results The mean survival time from group A to E were 7.20±2.93 days, 10.89±3.62 days, 10.50±3.40 days, 11.13±3.94 days and 12.80±4.07 days respectively. Significant difference was found among groups (F=3.20, P=0.022). The survival time at any group from B to E was significantly longer than group A(P value was 0.031, 0.047, 0.027, 0.001 respectively). No significant difference was found between group B and any group from C to E (P value was 0.815, 0.853, 0.255 respectively). No significant difference was found between group C and group D or group E (P value was 0.716, 0.161 respectively). No significant difference was found between group D and group E (P = 0.333). RI from group A to E were 7.80±1.48, 5.67±1.80, 6.10±1.66, 5.25±1.75 and 4.80±1.23 respectively. Significant difference was found between them (F=5.202, P=0.002). The RI at any group from B to E was lower than group A (P value was 0.005, 0.021, 0.002, 0.000 respectively). No significant difference was found between group B and any group from C to E(P value was 0.555, 0.592, 0.241 respectively). No significant difference was found between group C and group D or group E(P value was 0.265, 0.074 respectively). No significant difference was found between group D and group E (P=0.553). The cornea was obviously edematous, thickening, and showing cell infiltration in group A. The cornea cells showed layer disorder in group A. The degree of cornea infiltration was lighter in group B to E than in group A. Conclusion Cyclosporine A nino-particle eye drops of high, middle, low concentration and ordinary Cyclosporine A eye drops can effectively prevent cornea immune rejection after high risk cornea transplantation. (Ophthalmol CHN, 2012, 21: 116-120)

Key words: corneal transplantation, immune rejection, cyclosporine A, nano-particle eye drops

张海娟, 马科, 游玉霞, 吴雁, 徐清　. 环孢素A纳米粒滴眼液抑制大鼠高危角膜移植免疫排斥反应的实验研究[J]. 眼科, 2012, 21(2): 116-120.

ZHANG Hai-Juan, MA Ke, YOU Yu-Xia, WU Yan, XU Qing. Cyclosporine A nano-particle eye drops prevent the high risk corneal transplantation immune rejection in rats[J]. Ophthalmology in China, 2012, 21(2): 116-120.

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