摘要
目的探讨体外培养晶状体上皮细胞(LECs)的衰老情况以及衰老LECs对凋亡诱导因素的敏感性。方法体外培养兔LECs并传代。对第1、4、7代LECs进行研究,β-半乳糖苷酶组织化学染色检测衰老细胞,分析细胞衰老百分数的差异;原位末端标记法(TUNEL)检测培养过程中细胞的凋亡状况;10μmol/L喜树碱诱导细胞凋亡,比较各代细胞对喜树碱诱导凋亡敏感性的差异。结果LECs在体外培养过程中表现为有限生长的特性,经过增生、传代后衰老细胞增加。各代LECs在不受干预的情况下不发生凋亡。第1、4、7代LECs对喜树碱凋亡诱导因素的敏感性差异有统计学意义。结论LECs在体外培养过程中表现为有限生长的特性,增生的细胞以衰老为结局,衰老的LECs在不受外界干扰的情况下不发生凋亡,但对凋亡诱导因素的敏感性增加。
Objective Cataract is a senescent disease. It is also related to apoptosis of lens epithelial cells（ LECs）. The different defense abilities to apoptotic induction factor of senescent and unsenescent LECs in vitro was investigated in this paper. Methods LECs from 8 New Zealand white rabbits were cultured and passaged in DMEM containing 20% fetal bovine serum. Senescent cells were identified in the first,forth and seventh passage of confluence cells by x-gal staining. 10μmol/L camptothecin was used to induce cell apoptosis in different generations of lens epithelial cells to distinguish different sensitivities to apoptotic induction factor between senescent and unsenescent cells. Cell apoptosis was detected by TUNEL （ Roche）. Results The volume of senescent cells was enhanced,and multiple nuclei and vacule of cells were seen by x-gal staining. Apoptotic change was found in the camptothecin-treated cells. The senescent rate of the first, forth and seventh generation of LECs was （0.98±0.39）%,（24.98 ±3.55）%and（51.75 ±8.32）% respectively（F--215.22,P 〈0.01） and no apoptosis cell was found in simple culture group. In camptothecin-treated group, the apoptosis rate of LECs was （ 23. 87±3. 45 ） % , （ 32.08 ± 4.75 ） % and （ 38.29± 4. 01 ） % in different generations of cells （ F = 105.49, P 〈 0.01 ）, Senescent lens epithelial cells did not display spontaneous apoptosis. However,these cells were prone to die through apoptosis induced by camptothecin. Conclusion The passaged culture of LECs promote cellular senescence but do not induce cellular apoptosis. However, the vulnerabilities of senescence cells to apoptotic induction factor is higher than normal LECs.
出处
《眼科研究》
CSCD
北大核心
2008年第2期100-103,共4页
Chinese Ophthalmic Research
关键词
晶状体上皮细胞
衰老
凋亡
喜树碱
lens epithelial cells
cell senescence
cell apoptosis
camptothecin
作者简介
通讯作者：郭海科（Email：guohaike@medmail．com．cn）
