角膜干细胞端粒酶表达的实验研究-眼科学专业论文.docxVIP

!!!型垫=!：；兰l!堕堕兰堕坠堕坠塑匿堕———————————竖』兰l兰堡—!二角膜干细胞端粒酶表达的实验研究 !!!型垫=!：；兰l!堕堕兰堕坠堕坠塑匿堕———————————竖』兰l兰堡—!二 角膜干细胞端粒酶表达的实验研究 华中科技大学同济医学院附属协和医院眼科 研究生 陈宏 指导教师胡燕华教授 摘 要 目的检测角膜上皮各区域表层及基底层细胞端粒酶活性，探索 将端粒酶作为角膜缘干细胞标志物的可能性。方法采用TRAP方法 分别对8只新西兰大白兔角膜缘及中央透明角膜上皮细胞端粒酶活性 进行定性和定量分析。其中4只结膜下注射5--Fu 20mg／眼，4只未 注射任何药物：、f结果8只兔眼透明角膜上皮细胞平均生物发光值为 34912，端粒酶表达均为阴性；角膜缘上皮细胞平均生物发光值165575 端粒酶表达均为阳性，二者差异有显著性意义(P0．01)。5-Fu注射 组角膜缘上皮细胞平均生物发光值为185396，未注射5一Fu组角膜缘 上皮细胞平均生物发光值为145754，二者差异也有显著性意义 (P0．01)。阳性对照组生物发光值为210832，阴性对照组生物发光 值34793 0结论 角膜缘上皮细胞端粒酶表达阳性而透明角膜上皮细 胞表达阴性，角膜缘上皮基底层细胞的端粒酶活性显著高于表层L皮， 证明角膜缘存在具有极强增殖能力的干细胞，端粒酶的活性与干细胞 的分裂周期有关。端粒酶极有可能成为角膜干细胞的标志物。 关键词角膜缘干细胞端粒酶 兰主型垫查兰堕塑墨兰堕堕垦塑塑墨堕 兰主型垫查兰堕塑墨兰堕堕垦塑塑墨堕 竖主兰些堡苎 Detection of telomerase activity in corneal limbal stem cells Postgraduate Chen Hong Director Pr．Hu Yanhua ABS’I’RAC’I’ objective To investigate the possibility of using telomerase as a marker of corneallimbal stem cells． Method 8 rabbits corneal limbal tissues and 8 central corneal epithelial tissues were qualitativelly analyzed the telomerase activity by telomere repeat amplification protcol(TRAP)assay based on PCR；8 rabbits corneal tissues were quantitativelly analyzed by measuring light value，4 rabbits were injected 5-Fu subconjunctiva． Results 8 corneal limbal tissues were positive for telomerase activity and 8 central corneal tissues were negative．Telomerase activity of corneal limbal cells(1ight value 165575)was significantly higher than that of central corneal epithelial cells(1ight value 349 1 2)by telomerase quantitative examine technical(P0．01)．5-Fu injection group limbal cells show higher telomerase activity(1ight value 1 85396)than the group without injection(1ight value 145754)． Conclusions Positive telomerase activity was detected in the corneal limbal tissues．It suggests that there are stem cells in comeal limbus． Telomerase activity can be used as a marker of corneal limbal stem cells． Key words corneal limbus， stem cells，telomerase 2 }中科技大学同济医学院附属协和医院 }中科技大学同济医学院附属协和医院 博十学位论文 第一部分 角膜干细