摘要
目的：通过比较同为非选择性β受体阻滞剂的普萘洛尔和噻吗洛尔对小鼠血管瘤（EO-MA细胞）细胞体外增殖及凋亡的调控作用，初步探讨普萘洛尔和噻吗洛尔对小鼠血管瘤的治疗作用及可能机制，为临床应用β受体阻滞剂治疗婴幼儿血管瘤提供参考。方法体外培养EOMA细胞，取对数生长期EOMA细胞，随机分为药物组和对照组，用不同浓度普萘洛尔和噻吗洛尔作用EOMA细胞，分别干预24 h、48 h、72 h。通过连续光谱多功能酶标仪在570 nm和630 nm波长处测定以上质量浓度下光吸收值。应用四甲基偶氮唑盐比色法（MTT）检测细胞存活率、吖啶橙染色检测细胞凋亡情况，观察普萘洛尔和噻吗洛尔对EOMA细胞体外增殖和凋亡的影响。结果普萘洛尔作用24 h后，随剂量增加，EOMA细胞存活率逐渐下降，与对照组相比，至药物浓度为50μmol/L时有显著差异（P＜0.05），吖啶橙染色亦显示药物浓度为50μmol/L时凋亡细胞显著增多。48 h组和72 h组变化趋势与24 h组相似。噻吗洛尔作用24 h、48 h及72 h后，不同浓度和作用时间组之间，EOMA细胞存活率及凋亡率均无显著差异（P＞0.05）。结论普萘洛尔在体外可有效抑制小鼠血管瘤EOMA细胞的增殖，并促进其凋亡；而同为非选择性β受体阻滞剂的噻吗洛尔在体外对EOMA细胞的增殖和凋亡无显著调控作用。提示噻吗洛尔对小鼠血管瘤的作用可能与β受体通路以外的途径有关，其次，其治疗血管瘤的临床效果可能还需要多中心、大样本研究来验证。
Objetive To primarily study the possible treatment effects and its mechanism of timolol and propranolol on a Mouse Hemangioendothelioma Model （EOMA ）in vitro. Methods Comparative study on the effects of non-selective β-blocker propranolol and timolol,on the proliferation and apoptosis of Mouse He-mangioendothelioma Endothelial Cell （EOMA cells ）was conducted in vitro. EOMA cells were cultured in vitro,randomly divided into different groups,propranolol and timolol were added into the medium respectively, after 24,48,and 72 hours intervention,MTT assay and Acridine orange staining assay were conducted respec-tively to detect cell viability and apoptosis level. Results For propranolol,after 24 h treatment,significant differences of cell viability and apoptosis were noted （P＆lt;0.05）at the concentration of50 μmol/L,while con-tinuing to increase to 50 μmol/L,the cell survival rate decreased sharply to close to 0. Acridine orange stai-ning at the 50 μmol/L group after 24 h revealed many apoptotic cells.Groups of longer treatment to 48 h and 72 h showed a similar trend to the result of 24 h group in both MTT assay and Acridine orange staining. However,at the same time,compared with propranolol,timolol did not appear significantly inhibition on EOMA cell via-bility and induction on apoptosis in the experiment （P＆gt;0.05 ）. Conclusion Propranolol can effectively in-hibit the proliferation and induce the apoptosis of EOMA cells in vitro;While,timolol,also β-blocker,failed to play a similar role when added into the medium of EOMA cells in our study,showed no significant treatment potential,indicating that timolol’s treatment on EOMA cells may be related to pathways other than β-blocker way,furthermore,it is recommended to conduct further clinic study before its widely utilization in the treatment of infant hemangioma.
出处
《临床小儿外科杂志》
CAS
2014年第2期117-121,共5页
Journal of Clinical Pediatric Surgery
基金
本课题受国家自然科学基金项目（编号：81060267）、江西省自然科学基金项目（编号：2010GZY0016）、江西省教育厅青年基金项目（编号：GJJ11208）、赣州市科技项目及2012年赣南医学院本科生创新项目资助
