人纤溶酶原K5突变体滴眼液对大鼠角膜移植植片存活时间的影响

目的 探讨人纤溶酶原K5突变体(mK5)滴眼液预防大鼠角膜移植排斥反应发生的效果.方法 实验研究.以F344大鼠30只作为供体,Lewis大鼠60只作为受体,建立同种异体角膜移植排斥反应模型.另取15只Lewis大鼠行自体原位角膜移植,即A组为15只F344大鼠自体原位角膜移植.采用完全随机分组设计方案,将60只Lewis大鼠(60只右眼)分为B、C、D及E组.术后术眼4次/d滴眼液,每次1滴,A组和B组滴生理盐水,C组和D组滴mK5滴眼液,浓度分别为5 mg/L和10 mg/L,E组给予0.1%地塞米松滴眼液,连续用药14 d.根据Holland排斥反应评分标准,判断术后植片排斥情况.比较各组角膜植片的平均存活时间,并观察各时间点角膜新生血管生长情况,计算其面积.术后第14天,对各组大鼠角膜植片做组织学检查.采用方差分析和SNK-q检验对以上所得结果进行统计学分析.结果 B、C、D、E组植片平均存活时间分别为(9.3±2.1)、(21.1±7.3)、(23.5±10.8)及(28.2±19.1)d;C、D组分别与B组比较,差异有统计学意义(q=10.24,13.47:P<0.05);E组植片存活情况较C、D组好,但组间比较,差异无统计学意义(q=2.54,1.49;P>0.05).术后A、B、C、D及E组角膜新生血管开始出现的时间分别为(3.1±0.8)、(2.6±0.5)、(6.4±0.5)、(7.8±0.7)及(5.3±1.0)d;C、D、E组与A组比较(q=31.58,51.21,19.98;P<0.05);C、D、E组也较B组明显延长(q=43.87,67.14,24.53;P<0.05);C、D及E组两两组间比较差异有统计学意义(q=11.41,20.37,9.67;P<0.05).术后C、D组角膜新生血管的生长面积明显较B组少(q=30.76,62.14;P<0.05),且与E组比较差异有统计学意义(q=15.20,25.64;P<0.05).病理学检查显示,B组角膜植片可见大量炎性细胞浸润和角膜新生血管形成,而C组与D组植片炎性反应较轻,无明显新生血管形成.结论 mK5滴眼液可抑制同种异体大鼠角膜移植术后新生血管的生成,延缓角膜移植排斥反应的发生.

Effects of mutant kringle 5 eye drops on the survival time of corneal allografts in rat

Objective To investigate the preventative effect of mutant kringle 5 (mK5) eye drops on corneal allograft rejection. Methods It was a experimental study. The outbred strain F344 and Lewis rats were used as donors and recipients respeetively. Sixty Lewis rats were randomly divided into B, C, D and E Group;Group A, F344 rats autograft control;Group B, allograft control (the control groups were given normal sodium only);Group C and D, allograft groups, were treated with 5 mg/L and 10 mg/L mK5 eye drops respectively;Group E, allograft group, was treated with 0. 1% dexamethasone eye drops. The eye drops were applied one drop four times per day for two weeks, the occurrence and development of corneal allograft rejection and corneal neovascularization (CNV) was observed every other day by slit-lamp microscope,the grafts were evaluated clinically by means of Holland's scoring system and the area of CNV was calculated. Nine rats per group were killed on the 14th day, and the corneas were taken for histopathological examinations. Analysis of variance was used to analyze the outcomes. Results The average graft survival time of Group B, C, D and E was (9. 3 ± 2. 1 ) , ( 21.1 ± 7.3 ), ( 23.5 ± 10. 8 ) and ( 28.2 ± 19. 1 )d respectively,Compared with Group B, Group C and D had a statistically significant prolongation of survival time ( q = 10. 24,13.47 ;P < 0. 05 ). Though treated with 0. 1% dexamethnsone eye drops ( Group E) prolonged transplant survival time as compared with inK5 eye drops, but the difference was not statistically significant ( q = 2. 54,1.49;P > 0. 05 ). The occurrence of CNV in Group A was ( 3. 1 ± 0. 8 ) d, Group B ( 2. 6 ± 0. 5 ) d, Group C (6. 4 ± 0. 5 ) d, Group D (7. 8 ± 0. 7 ) d and Group E (5.3 ± 1. 0) d. Significant difference ( q = 31.58,51.21,19. 98 ;P < 0. 05 ) was found between groups C, D, E and Group A. There were also significant difference between groups C, D, E and Group B ( q = 43.87,67. 14,24. 53 ;P < 0. 05 ). The CNV areas of Group C and Group D were also smaller than Group B ( q = 30. 76, 62. 14 ;P < 0. 05 ). The results was similar compared with Group E ( q = 15.20,25.64;P < 0. 05 ). Fewer inflammatory cells and CNV were found in the cornea of the groups treated with mK5 eye drops. Conclusion Topical application of mK5 eye drops can prevent corneal graft rejection and corneal neovascularization in rats.