羟基喜树碱对青光眼滤过术后球结膜下成纤维细胞凋亡的影响及其机制研究  

The effects of hydroxycamptothecin on the apoptosis of fibroblasts isolated from subconjunctival scar in failed trabeculectomy and its mechanism

作　　者：汤伟[1] 钱朝旭[1] 袁志兰[1] 

机构地区：[1]南京医科大学第一附属医院(江苏省人民医院)眼科,210029

出　　处：《眼科》2012年第4期273-277,共5页Ophthalmology in China

摘　　要：目的观察羟基喜树碱(hydroxycamptothecin,HCPT)对青光眼滤过术失败的球结膜下瘢痕组织成纤维细胞凋亡的影响,并探讨其可能的作用机制。设计实验研究。研究对象球结膜下瘢痕组织成纤维细胞。方法体外培养青光眼滤过手术失败的球结膜下瘢痕组织成纤维细胞并鉴定。应用不同浓度的HCPT作用于成纤维细胞。采用流式细胞仪检测细胞凋亡。采用RT-PCR方法检测HCPT作用下,有或无广谱caspase阻断剂(Z-VAD-FMK)时,凋亡蛋白caspase-3、caspase-8、caspase-9 mRNA量的变化。主要指标凋亡蛋白caspase-3、caspase-8、caspase-9 mRNA量的变化。结果流式细胞检测显示,0.06、0.25、1.0、4.0 mg/l的HCPT作用于成纤维细胞24小时后,其凋亡率分别为0.87%、4.97%、5.76%、33.1%(P<0.05)。RT-PCR法显示,HCPT作用时caspase-3和caspase-9 mRNA的量较空白组明显增多,而caspase-8 mRNA的量较空白组减少;加入Z-VAD-FMK后,caspase-3、caspase-8、caspase-9 mRNA的量均减少。结论 HCPT在体外可诱导青光眼滤过手术球结膜下瘢痕组织成纤维细胞发生凋亡,且具有浓度依赖性,该凋亡可能通过caspase-3和caspase-9途径来实现。Objective To study the effect of hydroxycamptothecin（HCPT） on fibroblasts,which were isolated from the subconjunc- tiva scar in failed trabeculectomy and illustrate its possible mechanisms. Design Experimental study. Participants Fibroblasts cultured from the subconjunctival scar in failed trabeculectomy. Methods Fibroblasts cultured from the subconjunctival scar in failed tra- beculectomy were identified. Cell apoptosis was detected by flow cytometry （FCM）. Treated with different concentrations of HCPT, with or without caspase inhibitor Z-VAD-FMK, the expression levels of caspase-3, -8, -9 mRNA were observed with RT-PCR. Main Out- come Measures The expression levels of caspase-3, -8, -9 mRNA. Results FCM showed that the apoptosis rate of fiboblasts after 24 hours culture under 0.06, 0.25, 1.0, 4.0 mg/1 HCPT treatment was 0.87%, 4.97%, 5.76%, and 33.1%, respectively （P〈0.05）. By RT-PCR, the higher doses of HCPT were, the higher expression levels of caspase-3, and -9 mRNA were, while the expression level of caspase-8 was lower than the control group. When Z-VAD-FMK existed, it could decrease the levels of caspase-3, -8, -9 mRNA. Con- clusion HCPT can result in the apoptosis of fibroblasts in vitro and has dose-dependent. This apoptosis may be achieved through cas- pase-3, -9 routes. （Ophthalmol CHN, 2012, 21： 273-277）

关 键 词：羟基喜树碱 青光眼 成纤维细胞 凋亡 CASPASE-9 

分 类 号：R779.6[医药卫生—眼科]