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目的 检测晶状体诱导基因Pax6和Six3在晶状体摘除术后再生过程中的表达,探讨这两种基因与晶状体蛋白表达及细胞纤维化的关系.设计实验研究.研究对象48～53期非洲爪蟾蝌蚪.方法 实验组:选取发育正常的蝌蚪对其单侧的眼施以晶状体摘除手术;对照组:未做手术的另一侧正常眼.采用免疫荧光、蛋白印迹、qRT-PCR检测再生晶状体的眼睛中Pax6、Six3的表达情况.主要指标Pax6、Six3蛋白的表达水平.结果 与对照组相比,晶状体被摘除后随着时间推移逐渐再生.术后0～3天Pax6与Six3表达上升;术后3～14天,Pax6的表达先升高后降低,Six3表达下降,晶状体囊形成;术后14～21天,Pax6的表达持续降低,Six3的表达升高;术后21～30天,Pax6与Six3的表达下降,晶状体结构日趋完整.结论 晶状体再生的关键事件是部分角膜细胞去分化和晶状体蛋白表达.再生早期在Pax6和Six3的共同作用下外角膜内层细胞转分化形成晶状体囊,这两种基因也与后期晶状体蛋白表达及细胞纤维化的调控有密切关联.“,”Objective To detect the protein expression of the lens induction genes Pax6 and Six3 in lens regeneration after lentectomy,discussing the relation of these protein to crystallin expression and cell fibrosis.Design Experimental study.Participants Xenopus laevis tadpole at stage 48～53.Methods By immunofluorescence,Western blot and qRT-PCR were carried out to compare the expression of Pax6 and Six3 in lens regenerated eye and normal eye.Main Outcome Measures The expression of Pax6 and Six3 protein.Results Lens in the damaged eye gradually regenerates after lentectomy.Compared with the nomal eyes,the expression of Pax6 and Six3 is upregulated in 0～3 days after lentectomy.It was observed that Pax6 expression increased first and then gradually decreased while Six3 expression was found to decrease during 3～14 days after the lentectomy.Pax6 expression downregulated sequentially,but Six3 expression was found upregulated significantly during 14～21 days after lentectomy.Pax6 and Six3 expression downregulated during the interval of 21～30 days after the lentectomy.After this period,the lens regeneration process was found fundamentally completed.Conclusion In lens regeneration mechanism,the key event is the part of the cornea cell dedifferentiation and the expression of crystalline.Inner cell of outer cornea proliferation differentiation capsule by Pax6 and Six3 in initiation of regeneration,crystalline expression and cell fibrosis are closely associated with these genes in later period.