平阳霉素对血管瘤内皮细胞的作用

Effect of pingyangmycin on endothelial cells in infant hemangioma

目的探讨平阳霉素治疗血管瘤的作用机制。方法取2～18个月患儿增殖期血管瘤标本,原代培养血管瘤内皮细胞(HEC)。将不同浓度的平阳霉素(PYM)作用于3～5代生长旺盛的HEC,以MTT法筛选有效浓度。以有效浓度的PYM作用于HEC,光镜下观察不同时间的HEC形态,MTT检测细胞活性,流式细胞术检测细胞周期,Real-timePCR检测Shh的表达。结果PYM作用后,光镜下可见HEC发生弥散性坏死。MTT检测,吸光度降低;流式细胞术检测,S期细胞比例增多;Real-timePCR检测,Shh的表达明显降低。结论PYM可有效抑制HEC的生长和增殖,可能是通过抑制HECDNA的合成及抑制其分泌Shh,从而抑制内皮细胞的增殖、迁移及血管生成。

Objective To observe the mechanism of action of pingyangmycin （PYM） on hemangiomas endothelial cell （HEC） in proliferating hemangiomas in infant skin. Methods Samples came from 15 cases of proliferating hemangioma from infant skin （infants age 2 to 18 months, with 9 females and 6 males） and the hemangioma endothelial cells were primary cultured. Pingyangmycin of different density was added in the 3 rd- 5 th generation of endothelial cells, and the effective concentration（100 μg/ml） of pingyangmycin was selected by MTT assay. Pingyangmycin was added in the experimental group, PBS was added to the negative control group, and M199 medium was as control positive group. EC morphology was observed by inverted microscopy at 12, 24, 36 h, after addition of reagents. The cytcactivity and cell cycle were evaluated by MTT assay and flow cytometry. The expression of Shh was detected by real- time fluorescent quantitative polymerase chain reaction. Results Many HECs became necrotic after the addition of PYM, and there was much cell debris. The absorptance was gegraded by MTT assay, and FLC showed the percentage of S stage was increased. The expression of Shh was obviously diminished. Conclusion PYM can actively suppress the proliferation of hemangioma endothelial cells, and it may suppress DNA synthesis.Simultaneously, it may inhibit cellular secretion of Shh, and suppress HEC proliferation, migration and angiogenesis.