杜仲总黄酮对小儿血管瘤内皮细胞凋亡的影响

Efects of Eucommia ulmoides oliver flavones on apoptotic of pediatric hemangioma endothelial cells

目的研究杜仲总黄酮对小儿血管瘤内皮细胞凋亡的影响。方法体外培养小儿血管瘤内皮细胞,实验分为空白组(正常培养小儿血管瘤内皮细胞);杜仲总黄酮高剂量组(50μg/mL杜仲总黄酮处理小儿血管瘤内皮细胞);杜仲总黄酮中剂量组(5μg/mL杜仲总黄酮处理小儿血管瘤内皮细胞);杜仲总黄酮低剂量组(0.5μg/mL杜仲总黄酮处理小儿血管瘤内皮细胞),继续培养72 h后,MTT法检测细胞活力;TUNEL法检测细胞凋亡;Western blot法检测小儿血管瘤内皮细胞凋亡相关蛋白Bax、Bcl-2和caspase-9表达水平。结果与空白组相比,杜仲总黄酮中、高剂量组能够抑制小儿血管瘤内皮细胞增值,差异有统计学意义(P<0.05);杜仲总黄酮中、高剂量组细胞凋亡升高,差异有统计学意义(P<0.05);杜仲总黄酮高剂量组Bax蛋白表达上升,差异有高度统计学意义(P<0.01);杜仲总黄酮中、高剂量组caspase-9蛋白表达上升,差异有统计学意义(P<0.05);杜仲总黄酮低、中、高剂量组Bcl-2蛋白表达降低,差异有统计学意义(P<0.05)。结论杜仲总黄酮能够通过抑制Bcl-2蛋白表达、促进Bax和caspase-9蛋白表达,从而诱导小儿血管瘤内皮细胞发生凋亡,有望成为是一种有效的小儿血管瘤治疗剂。

Objective To study the influence of Eucommia ulmoides oliver flavones（EUOF） on apoptosis of infantile hemangioma endothelial cell. Methods Infantile hemangioma endothelial cells were cultured in vitro. The cells was divided into blank groups（normal cultivate infantile hemangioma endothelial cells）, high dose group of EUOF（50 μg/mL EUOF treatment infantile hemangioma endothelial cells）, the middle dose group（5 μg/mL EUOF treatment infantile hemangioma endothelial cells）, the low dose group（0.5 μg/mL EUOF treatment children hemangioma endothelial cells）,then cultured for 72 hours, MTT method detected cell vitality. The TUNEL method detected apoptosis of cells; Western blot method detected expression levels of apoptosis related proteins Bax, Bcl-2 and caspase-9 in infantile hemangioma endothelial cell. Results Compared to the blank group, infantile hemangioma endothelial cell proliferation could inhibit in EUOF middle dose group, high dose group, the differences were statistically significant（P〈0.05）. The rate of apoptosis in high dosages group and middle dosages group rose, the differences were statistically significant（P〈0.05）; the expression of Bax in high dose group rose, differences were statistically significant（P〈0.01）. The expression of caspase-9 potein in the high dose group middle dose group and low dose group rose, the differences were statistically significant（P〈0.05）. The expression of bcl-2 in high dose group, middle dose group decreased, the differences were statistically significant（P〈0.05）. Conclusion EUOF can inhibit the the expression of Bcl-2 protein and promote the expression of Bax and caspase-9 in infantile hemangioma endothelial cells, that is expected to be an effective therapeutic agent in the disease of hemangioma.