三种不同β受体阻滞剂治疗小鼠血管瘤的实验研究  

Experimental study of three different β-receptor blockers in treatment on murine hemangioendothelioma

作　　者：徐仙赟[1] 谢琼珺[1] 彭威[1,2] 宁慧婷[1,2] 温俊程 王涛[1] 马靓雯 郝彦斌[3] 刘潜[1] 

机构地区：[1]江西省脉管性疾病临床研究中心(培育),赣州341000 [2]江西省赣南医学院研究生院,赣州341000 [3]江西省赣南医学院基础医学院,赣州341000

出　　处：《中国免疫学杂志》2016年第8期1118-1122,共5页Chinese Journal of Immunology

基　　金：江西省赣鄱英才"555工程"领军人才项目(赣才字[2012]1号);人力资源与社会保障部2011年度留学人员科技启动项目(赣人社字[2011]474号);江西省自然科学基金项目(No.2010GZY0016);江西省青年自然科学基金项目(No.20142 BAB215026);江西省教育厅教学改革项目(No.JXJG-14-13-11);江西省卫生与计划生育委员会中医药科研项目(No.2014A149);赣南医学院自然科学研究计划(No.YB201312);2015年赣南医学院本科生创新项目(ZR2014015;ZR2014016和ZR2014017)资助;江西省教育厅科学技术项目(No.150963)

摘　　要：目的:比较三种不同β受体阻滞剂对小鼠血管瘤(EOMA细胞)细胞体内外的影响,初步探讨普萘洛尔对小鼠血管瘤的治疗作用及可能机制,为β受体阻滞剂治疗婴幼儿型血管瘤提供参考。方法:体外培养EOMA细胞,用不同浓度美托洛尔、普萘洛尔、布托沙明作用EOMA细胞,作用24 h。应用MTT法检测细胞存活率、吖啶橙染色检测细胞凋亡情况,观察美托洛尔、普萘洛尔、布托沙明对EOMA细胞体外增殖和凋亡的影响。体内实验将EOMA细胞移植裸鼠,待肿瘤体积生长至100 mm3,将动物随机分为4组,分别为对照组、美托洛尔组、布托沙明组和普萘洛尔组,药物组按2 mg/(kg·d)灌胃给药,对照组给予等体积的生理盐水(NS),每两天测量肿瘤体积大小。实验结束时收集血清,酶联免疫吸附实验检测血清中肿瘤坏死因子-α(TNF-α)和血管内皮细胞生长因子(VEGF)的表达水平。结果:体外实验显示,普萘洛尔作用24 h后,EOMA细胞存活率随剂量增加逐渐下降,50μmol/L时有显著差异(P<0.05),药物增加至800μmol/L,细胞存活率接近10%。吖啶橙染色显示50μmol/L时凋亡细胞显著增多。美托洛尔、布托沙明作用24 h后,100μmol/L时细胞存活率与对照组相比有显著差异(P<0.05),800μmol/L时细胞存活率下降至20%,明显高于相同浓度的普萘洛尔组,存在显著性差异(P<0.05)。吖啶橙染色亦出现相似的规律。体内实验显示,实验结束时美托洛尔、布托沙明小鼠和普萘洛尔药物组的肿瘤体积分别为(1 642.8±89.3)、(1 529.3±119.1)和(752.7±46.5)mm3,与对照组小鼠的肿瘤体积(2 023.3±123.07)mm3相比明显减少(P<0.001)。美托洛尔、布托沙明小鼠和普萘洛尔药物组血清中VEGF分别水平为(606.5±105.8)pg/ml、(534.3±243.2)pg/ml和(420.1±123.7)pg/ml,明显低于PBS对照组的[(825.8±145.7)pg/ml,(P<0.05)],TNF-α结果依次是(301.3±62.3)pg/ml、(305.1±53.8)pg/ml和(288.8±59.5)pg/ml,明显低于正常对照组[(444Objective： To comparison of three different beta blockers on murine hemangioma（ EOMA cells） cells in vitro and in vivo effects. Preliminary study on the therapeutic effect of propranolol on vascular tumor in mice and possible mechanisms,provide a reference for beta blockers in the treatment of infantile hemangioma. Methods： Comparative study on the effects of three kinds of differentβ-receptor blockers---metoprolol,propranolol and butoxamine,on the proliferation and apoptosis of Mouse Hemangioendothelioma 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 h intervention. MTT assay and acridine orange staining assay were conducted respectively to detect cell viability and apoptosis level. EOMA cells were transplanted into nude mice in vivo. Tumor volume growth to 100 mm^3,animals were randomly divided into 4 groups respectively,the control group,metoprolol group,Bhutto Samin group and propranolol group,drug group according to 2 mg /（ kg·d） oral gavage,control group were given an equal volume of saline（ NS）,every two days measurement tumor volume size. Serum levels of tumor necrosis factor alpha（ TNF-α） and vascular endothelial growth factor（ VEGF） were detected by enzyme linked immunosorbent assay（ ELISA） in the end of the experiment. Results： For propranolol,after 24 h treatment,significant differences of cell viability and apoptosis were noted（ P〈0. 05） at the concentration of 50 μmol / L,while continuing to increase to 800 μmol / L,the cell survival rate decreased sharply to close to 10%. Acridine orange staining at the 50 μmol / L group after 24 h revealed many apoptotic cells. For metoprolol and butoxa mine,significant differences of cell viability and apoptosis were noted（ P〈0. 05） at the concentration of 100 μmol / L,while continuing to increase to 800 μmol / L,the cell survival rate decreased sharply to clo

关 键 词：小鼠血管瘤 Β受体阻滞剂 普萘洛尔 凋亡 TNF-Α 

分 类 号：R392.7[医药卫生—免疫学]