自家血治疗实验性兔角膜溃疡后对角膜中基质金属蛋白酶2、基质金属蛋白酶组织抑制剂-2的表达影响

The effects of autologous blood on the expression of matrix metalloproteinase 2 and tissue inhibitor of metalloproteinase-2 levels in rabbits with experimental corneal ulcer

目的:观察自家血治疗实验性兔角膜溃疡的疗效,并探讨自家血对溃疡角膜中基质金属蛋白酶(Matrix Metalloproteinase,MMP)2及基质金属蛋白酶组织抑制剂-2(Tissue Inhibitor of Metalloproteinase-2,TIMP-2)表达的影响和作用。方法:将36只实验兔随机分为A、B、C、D四组,每组9只。B、C、D组通过角膜环钻在兔角膜中央处钻取一个直径5 mm、深度0.1 mm的角膜溃疡灶,建立动物模型。自术后第1天开始,B、C、D组采用盐酸左氧氟沙星滴眼液点眼每日4次,作为抗感染基础治疗;B组采用自家血点眼每日2次;C组采用速高捷眼用凝胶点眼每日3次;D组采用生理盐水点眼每日3次。术后第3、7、14天行角膜炎症和溃疡评分,并于术后第14天从各组随机抽取三只角膜应用免疫印迹法检测角膜中MMP2、TIMP-2的表达。结果:角膜评分显示,术后第3天B、C、D三组间的角膜炎症评分和溃疡评分差异均无统计学意义(均为P>0.05);B组和C组的角膜炎症和溃疡评分在术后第7天起各时间点均小于D组(均为P<0.05);术后第7天B组的角膜炎症和溃疡评分低于C组(均为P>0.05);术后第14天B组的角膜炎症和溃疡评分低于C组(均为P<0.01)。免疫印迹法结果显示,术后第14天,MMP2的表达,B、C组均明显低于D组(均为P<0.01),B组明显低于C组(P<0.01)。TIMP-2的表达,B、C组均明显高于D组(均为P<0.01),B组明显高于C组(P<0.01)。结论:自家血治疗实验性兔角膜溃疡的疗效显著。自家血点眼治疗实验性兔角膜溃疡是通过提高角膜中TIMP-2的表达,抑制MMP2的表达,从而减轻角膜溃疡中基质层的损伤程度,缩短病程,促进其愈合。

Objective:To investigate the efficacy of autologous blood on experimental corneal ulcer in rabbits,and explore the effects of autologous blood on the expression levels of MMP2 and TIMP-2 in ulcerated cornea.Methods:A total of 36 experimental rabbits were randomly divided into four groups,namely group A,group B,group C and group D,with 9 rabbits in each group.In group B,group C,and group D,a corneal ulcer with a diameter of 5 mm and a depth of 0.1 mm was drilled at the center of the cornea of each experimental rabbit by a corneal trephine to establish animal models.From the first day after operation,groups B,group C and group D were treated with levofoxacin hydrochloride eye drops 4 times a day as the basic anti-infection treatment.In addition,group B was instilled with autologous blood,twice a day;group C was instilled with the Sugaojie ophthalmic gel(速高捷眼用凝胶),three times a day,and group D was treated with normal saline,three times a day.The corneal infammation and ulceration scores were performed on the 3rd,7th and 14th day after operation.Three corneas were randomly selected from each group on the 14th day after operation,and the expression levels of MMP2 and TIMP-2 in the corneas were detected by Western blot.Results:The corneal score showed that there was no significant difference in corneal infammation score and ulceration score among groups B,group C and group D on the third day after operation(all P>0.05).The scores of corneal infammation and ulceration in group B and group C were lower than those in group D at each time point from the 7th day after operation(all P<0.05).On the 7th day after operation,the scores of corneal infammation and ulceration in group B were lower than those in group C(all P>0.05).On the 14th day after operation,the scores of corneal infammation and ulceration in group B were lower than those in group C(all P<0.01).The result of Western blot showed that the expression of MMP2 in group B and group C were significantly lower than that in group D(all P<0.01),and that in group B was significantly lower than that in group C(P<0.01).The expression of TIMP-2 in group B and group C were significantly higher than that in group D(all P<0.01),and that in group B was significantly higher than that in group C(P<0.01).Conclusion:The effect of autologous blood on experimental corneal ulcer in rabbits is significant.In the treatment of experimental rabbit corneal ulcer by autologous blood spotting,it can increase the expression of TIMP-2 and inhibit the expression of MMP2 in the cornea,there by reducing the damage degree of stromal layer in the corneal ulcer,shortening the course of the disease,and promoting its healing.