去整合素抑制兔晶状体上皮细胞增殖的实验研究

Experimental study on Kistrin inhibiting proliferation of lens epithelial cells in rabbit

目的探讨去整合素（Kistrin）对兔眼后发性白内障发生过程中晶状体上皮细胞（lens epithelial cells,LEC）增殖的抑制作用。方法雄性新西兰大白兔36只,随机分为对照组：A组,实验组：B、C、D组,每组各9只兔9眼（右眼）。4组均行透明晶状体囊外摘出术,术毕A组兔眼前房注入林格液0.2mL,B、C、D组分别注入40μg.L-1、80μg.L-1和160μg.L-1Kistrin溶液0.2mL。术后第14天采用免疫组织化学法对4组兔后发性白内障发生过程中LEC增殖细胞核抗原（proliferating cell nuclearantigen,PCNA）的表达进行检测,计算增殖指数（proliferative index,PI）值,PI=PCNA阳性细胞数/50×100%;通过HE染色及透射电镜观察角膜改变情况。结果（1）术后第14天PI值A组：0.320±0.020、B组：0.175±0.030、C组：0.065±0.020、D组：0.190±0.050,B、C、D组较A组PI值均明显降低（均为P〈0.01）;C组较B、D组PI值均明显降低（均为P〈0.01）,B组与D组PI值比较差异无统计学意义（P〉0.05）;（2）术后第14天赤道部PI值A组：0.390±0.060、B组：0.230±0.040、C组：0.110±0.030、D组：0.240±0.080;中央部PI值A组：0.250±0.050、B组：0.120±0.030、C组：0.060±0.020、D组：0.140±0.040,4组赤道部较中央部PCNA阳性表达均明显升高（均为P〈0.01）。（3）实验组C组与A组比较：角膜内皮细胞超微结构无明显改变。结论40~160μg.L-1的Kistrin灌注液可有效地抑制兔眼后发性白内障的发生;赤道部LEC增殖能力强于中央部;80μg.L-1可作为进一步研究Kistrin抑制后发性白内障形成的较适宜浓度。

Objective To explore the inhibition of Kistrin on proliferation of lens epithelial cells（LEC）in the process of after-cataract in rabbits.Methods Thirty-six male New Zealand rabbits were randomly divided into control group（group A）and experimental groups（group B,group C and group D）,9 rabbits（9 eyes,right eye）in each group.All rabbits in each group were treated with extracapsular lens extraction surgery.After the surgery,0.2 mL Ringer＇s solution was injected into anterior chamber of rabbits in group A,40 μg·L-1,80 μg·L-1 and 160 μg·L-1 Kistrin solution for 0.2 mL were injected into chamber of rabbits in group B,group C and group D,respectively.The expression of proliferating cell nuclear antigen（PCNA）was detected in the process of after-cataract for rabbits of 4 groups by immunohistochemical methods at the 14th day after surgery.The proliferative index（PI）value was calculated（PI=positive cell number of PCNA/50×100%）.Furthermore,corneal and retinal changes were observed by HE staining and transmission electron microscopy.Results PI values were 0.320±0.020,0.175±0.030,0.065±0.020 and 0.190±0.050 in group A,group B,group C and group D at the 14th day after surgery,respectively.The PI values of group B,group C and group D were obviously lower than that of group A（All P0.01）;The PI value of group C was significantly lower than those of group B and group D（Both P0.01）;The PI value of group B had no statistical difference with that of group D（P0.05）.The PI values at ambitus were 0.390±0.060,0.230±0.040,0.110±0.030 and 0.240±0.080 in group A,group B,group C and group D at the 14th day after surgery,respectively;The PI values at central part were 0.250±0.050,0.120±0.030,0.060±0.020 and 0.140±0.040 in group A,group B,group C and group D at the 14th day after surgery,respectively;The positive expression of PCNA at ambitus was significantly higher than that at central part（P0.01）.The ultrastructure of corneal endothelial cells in group C had no significant difference with group A.Conclusions After-cataract of rabbit eyes can be effectively inhibited by 40 μg·L-1 to 160 μg·L-1 Kistrin;The proliferative effect of LEC in ambitus is stronger than that in central part;And 80 μg·L-1 can be a suitable concentration for Kistrin in studying its inhibitive effects on after-cataract.