康柏西普对豚鼠形觉剥夺性近视的干预作用及相关机制分析

目的　探讨抗新生血管生长因子融合蛋白康柏西普（Conbercept）球结膜下注射对豚鼠形觉剥夺性近视的干预作用及其可能的作用机制。方法　普通级3周龄断乳花色豚鼠48只随机分为空白对照组、单纯手术组、生理盐水组、Conbercept组，每组各12只。其中空白对照组豚鼠不作处理，单纯手术组、生理盐水组、Conbercept组使用眼睑缝合法遮盖右眼，生理盐水组、Conbercept组分别在缝合时、缝合后7 d右眼球结膜下注射0.05 mL生理盐水、0.5 mg（0.05 mL）Conbercept。记录各组眼睑缝合前及缝合后7 d、14 d时的眼轴长度、屈光度，并取后极部巩膜组织采用RT-PCR、Western blot方法检测各组豚鼠巩膜中基质金属蛋白酶2（MMP-2）、基质金属蛋白酶抑制剂-2（TIMP-2）、转化生长因子（TGF）-β1、TGF-β2 mRNA和蛋白表达情况。结果　眼睑缝合后7 d、14 d，单纯手术组较空白对照组豚鼠右眼眼轴长度、屈光度均增加，MMP-2、TGF-β2 mRNA和蛋白相对表达量均增加，TGF-β1、TIMP-2相对表达量均减少，差异均具有统计学意义（均为P<0.05）；单纯手术组与生理盐水组各指标相比差异均无统计学意义（均为P>0.05)。眼睑缝合后7 d，与生理盐水组相比，Conbercept组眼轴长度、屈光度及TIMP-2、TGF-β1 mRNA和蛋白相对表达量差异均无统计学意义（均为P>0.05），MMP-2、TGF-β2 mRNA和蛋白相对表达量均减少，差异均有统计学意义（均为P<0.05）；眼睑缝合后14 d时，与生理盐水组相比，Conbercept组眼轴长度、屈光度及MMP-2、TGF-β2、TGF-β1 mRNA和蛋白相对表达量差异均有统计学意义（均为P<0.05）。结论　0.5 mg Conbercept球结膜下注射可延缓豚鼠形觉剥夺性近视形成，且可能是通过调节巩膜中的MMP-2、TGF-β1、TGF-β2表达而发挥作用。

Effects of Conbercept on the form-deprivation myopia in guinea pigs and its related factors

Objective　To investigate the effect of subconjunctival injection of anti-angiogenic growth factor fusion protein, Conbercept, on the form-deprivation myopia in guinea pigs and its underlying mechanisms.Methods　All of 48 healthy guinea pigs (male or female) of 3 weeks old were taken. The guinea pigs were randomly divided into control group, unilateral lid fusion group, normal saline solution group and Conbercept group. Guinea pigs in the control group left untreated, and the unilateral lid fusion group, normal saline solution group and Conbercept group were treated to cover the right eye with eyelid suture. The right eye were performed with subconjunctival injection of 0.05 mL normal saline solution in the normal saline solution group, while 0.5 mg (0.05 mL) Conbercept for the Conbercept group at the time of eyelid suture and 7days after eyelid suture. Then the diopter and axial length were recorded before and 7 days and 14 days after injection, and the sclera was collected at the posterior pole for RT-PCR and Western blot to detect mRNA and protein expression levels of matrix metalloproteinases-2 (MMP-2), tissue inhibitor of matrix metalloproteinases-2 (TIMP-2), and transforming growth factor-β (TGF-β).Results　Seven days and 14 days after eyelid suture, when compared with the control group, the length of the eye axis and diopter of the right eye of guinea pigs in the unilateral lid fusion group were increased; the relative expression levels of MMP-2, TGF-β2 mRNA and protein were increased, and the relative expression levels of TGF-β1 and TIMP-2 were decreased, and the differences were statistically significant (all P<0.05). There were no statistically significant differences in all variables between the unilateral lid fusion group and normal saline solution group (all P>0.05). Seven days after eyelid suture, compared with the normal saline solution group, there were no statistically significant differences in the axial length, diopter, and relative expression of TIMP-2, TGF-β1mRNA and protein in the Conbercept group (all P>0.05), but the relative expression levels of MMP-2 and TGF-β2mRNA and protein decreased, and the differences were statistically significant (both P<0.05). At 14 days after eyelid suture, compared with normal saline solution group, the axial length, diopter and relative expression levels of MMP-2, TGF-β2, TGF-β1mRNA and protein in the Conberpt group were statistically significant (all P<0. 05). Conclusion Subconjunctival injection of 0.5 mg Conbercept can delay the formation of form-deprivation myopia in guinea pigs, which is achieved possibly by regulating the expression of MMP-2, TGF-β1 and TGF-β2 in the sclera. Conclusion　Subconjunctival injection of 0.5 mg conbercept can delay the formation of form-deprivation-myopia in guinea pigs,possibly by regulating the expression of MMP-2,TGF-β1 and TGF-β2 in the sclera.