Abstract: 【Abstract】 Objective To study the effect of hydroxycamptothecin(HCPT) on fibroblasts，which were isolated from the subconjunctiva 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 trabeculectomy 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 Outcome 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/l 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. Conclusion HCPT can result in the apoptosis of fibroblasts in vitro and has dose-dependent. This apoptosis may be achieved through caspase-3, -9 routes. (Ophthalmol CHN, 2012, 21: 273-277)

Key words: hydroxycamptothecin, glaucoma, fibroblasts, apoptosis, caspase-9