摘要
目的研究重组人血管内皮抑索（YH-16，恩度）对鼠源性血管瘤内皮细胞（EOMA）的抑制作用及其机制，并与血管瘤常用药物曲安缩松、平阳霉素（PYM）、干扰素α-2a相比较，观察单用及联合用药对EOMA细胞的抑制作用。方法用MTT法检测不同浓度的YH-16对EOMA细胞作用24、48、72h的抑制率，筛选出YH-16的最适抑制浓度。比较YH-16、曲安缩松、PYM、干扰素α-2a四种药物单用以及YH-16、曲安缩松、平阳霉素两两联合作用对EOMA细胞的抑制率。采取流式细胞术测定YH-16作用EOMA细胞48h后，细胞凋亡率及Caspase-3的活性。结果①YH-16各浓度组在24h、48h及72hi个作用时段对EOMA细胞抑制作用显著（P〈0．01），并存在明显的剂量依赖关系；②选择200ptg／ml为YH-16的最适抑制浓度；③四种药物抑制作用由强到弱：PYM〉YH-16〉曲安缩松〉干扰素α-2a；④YH-16与曲安缩松或与PYM的联合效应均为拮抗效应，Q值均〈0．85；PYM与曲安缩松的联合效应为相加效应，0．85〈Q值〈1．15；⑤YH-16作用后细胞凋亡率随药物浓度增加而增高，Caspase-3表达在YH-16各浓度组均增强（P〈O．01）。结论YH-16对EOMA细胞有确切的抑制作用。该作用弱于PYM，而强于曲安缩松及干扰素α-2a，它主要是通过诱导细胞凋亡实现，且Caspase-3参与此过程。YH-16能拮抗曲安缩松及PYM抑制EOMA细胞增殖的作用，曲安缩松能增加PYM对EOMA的抑制增殖作用。
Objective To investigate the mechanism of inhibitory effects of rh-endostatin （YH- 16, endostar） on the proliferation of murine hemangioendothelioma cell line（EOMA） in vitro, compare its inhibitory effects with Triamcinolone Acetonide, Pingyangmycin （PYM）, interferon alpha 2a, and observe the combined inhibitory effects on the growth of EOMA cells in vitro. Methods The IR of EOMA cells managed by different concentration of YH-16 for 24 h, 48 h and 72 h were compared by MTT assay to get an optimal concentration. MTT assay was also used to detect the combined effect of any two of YH-16. Triamcinolone Acetonide or PYM. The changes of apoptosis and Caspase-3 protein expression were examined by flow cytometry （FCM） after YH-16 treatment for 48 h. Results YH-16 with different concentration had significant inhibitory effect on proliferation of EOMA cells at 24 h, 48 h and 72 h（P〈0.01）,and IR and the concentration had significant dose-effect relationship. The opti- mal concentration of YH-16 was 200 htg/ml. The inhibitory effects from the strongest to the weakest was as follows： PYM〉 YH-16 〉 Triamcinolone Acetonide〉 interferon alpha 2a. Q value of YH-16 combined with Triamcinolone Acetonide or PYM was lower than 0. 85, but Q value of Triamcinolone Acetonide combined with PYM was between 0. 85 and 1.15. The apoptosis rate of EOMA cells treated with YH-16 for 48h was also concentration-dependant. The expression of Caspase 3 increased in all groups with different concentration （P〈0. 01 ）. Conclusions YH-16 can effectively inhibit the growth of EOMA cells. Its inhibitory effect is stronger than that in Triamcinolone Acetonide and interferon alpha 2a, but weaker than PYM. The inhibitory mechanism of YH-16 is to induce apoptosis with the participation of Caspase-3. YH-16 can be an antagonism to Triamcinolone Acetonide and PYMs in the inhibition of EOMA proliferation, and the combination of Triamcinolone Acetonide and PYM has an effect of addition.
出处
《中华小儿外科杂志》
CSCD
北大核心
2008年第8期475-478,共4页
Chinese Journal of Pediatric Surgery
