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为进一步调控角膜基质细胞周期、抑制角膜基质细胞增殖提供基础和靶点 ,检测原位人角膜基质细胞和体外培养的人角膜基质细胞是否表达细胞周期蛋白 (cyclin)—细胞周期蛋白依赖性激酶 (cyclin dependentkinase,CDK)—细胞周期蛋白依赖性激酶抑制因子 (cyclin dependent kinase inhibitor,CKI)网络成员。方法 :制作人角膜冰冻切片 ,并体外培养人角膜基质细胞 ,分别用免疫组织化学和免疫细胞化学的方法检测其中 cyclin D、E,cdk2、4,p2 1WAF1的表达。结果 :体外培养的人角膜基质细胞中有 cyclin D、E,cdk2、 4,p2 1WAF1蛋白水平的阳性表达 ,而原位人角膜基质细胞中无阳性表达。结论 :体外培养的人角膜基质细胞有别于静止状态的角膜基质细胞 ,与在体激活的角膜基质细胞相似 ,cyclin—CDK—CKI成员在其中有阳性表达的结果 ,为进一步通过人为下调 cyclin/ CDK水平、上调 CKI水平以抑制基质细胞增殖奠定了基础
To provide a basis and target for further regulation of corneal stromal cell cycle and inhibition of corneal stromal cell proliferation, whether human corneal stromal cells in situ and cultured human corneal stromal cells express cyclin-cyclin-dependent kinase cyclin dependent kinase (CDK) - a member of the cyclin dependent kinase inhibitor (CKI) network. Methods: Human cornea frozen sections were made and human corneal stromal cells were cultured in vitro. The expressions of cyclin D, E, cdk2, 4 and p2 1 WAF1 were detected by immunohistochemistry and immunocytochemistry respectively. Results: There was positive expression of cyclin D, E, cdk2, 4 and p2 1WAF1 protein in cultured human corneal stromal cells, but no positive expression in in situ human corneal stromal cells. CONCLUSION: Human corneal stromal cells cultured in vitro are different from resting corneal stromal cells. Similar to in vivo activated corneal stromal cells, cyclin-CDK-CKI members are positively expressed in human corneal stromal cells. CDK levels, upregulation of CKI levels to inhibit stromal cell proliferation laid the foundation