HCPT联合Ver诱导兔结膜成纤维细胞的凋亡实验研究  

Experimental research of HCPT and Ver to induce apoptosis of rabbit's conjunctival fibroblast

作　　者：毕燕龙 荣翱[2] 胡桃舫[2] 李锦文[2] 

机构地区：[1]上海铁路中心医院眼科,上海市200072 [2]同济大学附属同济医院眼科

出　　处：《眼科新进展》2002年第1期18-20,共3页Recent Advances in Ophthalmology

基　　金：铁道部科研基金项目(编号 :TDB)~~

摘　　要：目的　观察羟基喜树碱 (hydroxycam ptothecin,H CPT)、维拉帕米 (verapam il,Ver)单独或联合应用诱导兔体外培养成纤维细胞 (fibroblast ,FB)凋亡的作用。方法　采用兔结膜 FB体外原代培养及 TU NEL染色法观察 HCPT单独或联合 Ver应用时 ,对兔结膜 FB凋亡指数 (apoptotic index,A I)、形态的影响 ,并进行相关统计学分析。结果　 H CPT 1m g· L- 1作用 12 h后 ,细胞开始出现凋亡的形态学改变 ,5～ 3 0 m g· L - 1的 HCPT诱导 FB的 A I高峰均出现在作用后 72 h左右 ,且随着浓度增加 ,凋亡细胞数增多。5m g· L- 1H CPT联合 5m g· L- 1Ver诱导 FB的 A I(48h为 2 4 .5± 3 .4 )于 12～ 4 8h略低于 10 m g· L- 1组 H CPT的 A I (48h为 2 4 .6± 3 .9) ,当作用时间达 72 h时 ,则前者的 A I (3 0 .4± 3 .4 )大于后者的 AI (2 5.2± 5.6) (χ2 =6.74 ,P<0 .0 1)。结论　 H CPT可诱导兔 FB凋亡 ,当联合 V er应用时 ,诱导凋亡作用增强。Objective To observe the induction of apoptosis of fibroblast in vitro by hydroxycamptothecin (HCPT) and/or verapamil (Ver) .Methods Fibroblasts were obtained and cultured directly from rabbit's conjunctiva. The third to fifth generations of fibroblasts were used , cultured together with HCPT and Ver using different concentrations and different time durations. We applied TUNEL analyzing methods to observe apoptotic index (AI) and shape changes of fibroblasts. Results Morphologic changes of apoptosis began to appear after 1mg·L -1 ·12h -1 HCPT , the fastigium of AI of 5~30mg·L -1 HCPT were all evident after about 72h. And with increase of concentrations of HCPT, the AI also showed some trends of elevation. When HCPT and Ver were used together, the function of AI elevation was greater than HCPT in the same concentration ( P <0.01).Conclusion HCPT can induce apoptosis to fibroblasts, this function can be increased when combined with Ver.

关 键 词：成纤维细胞 羟基喜树碱 维拉帕米 凋亡 

分 类 号：Q939.836[生物学—微生物学] R77[医药卫生—眼科]