摘要
目的:研究不同浓度的镍对体外培养人脐静脉内皮细胞(HUVEC)的影响。方法:用两种镍化合物(NiCl2·6H2O、NiSO4·6H2O)分别制成含Ni(Ⅱ)浓度为1000、500、250、125、62.5μmol/L及31.25μmol/L的溶液,作用于体外培养的HUVEC细胞株,72h后MTT法测试细胞增殖活性,确定NiCl2·6H2O与NiSO4·6H2O的TC50,并分别以TC50的Ni(Ⅱ)作用培养的HUVEC细胞株,MTT法检测不同时间点(24h、48h、72h、96h、120h)的细胞相对增殖活性;2,4-二硝基苯肼法检测作用72h时培养上清液及细胞裂解液中乳酸脱氢酶(LDH)的含量,计算Ni(Ⅱ)化合物作用下细胞LDH的释放率,评价Ni(Ⅱ)对其影响;HE常规染色观察两种镍化合物作用72h后的细胞形态;AnnexinV-FITC/PI双标记流式细胞仪检测细胞凋亡。结果:两种化合物作用HUVEC细胞TC50为125μmol/L,随着浓度的不断增加,细胞毒性增强;同一浓度的Ni(Ⅱ)对细胞活性的影响随着作用时间增加而增强;两种镍化合物对细胞LDH释放率的影响无明显差异(P>0.05);NiSO4·6H2O对细胞形态及凋亡率的影响显著大于NiCl2·6H2O(P<0.05)。结论:Ni(Ⅱ)对HUVEC细胞毒性影响呈时间、浓度依赖性,NiSO·46H2O对HUVEC细胞形态及凋亡影响大于NiCl·26H2O。
Objective：To investigate the effects of nickel on cultured human umbilical vein endothelial cells（HUVEC） in different concentrations. Methods：Ni（Ⅱ） concentration of 1000 μmol/L,500 μmol/L,250 μmol/L,125 μmol/L,62.5 μmol/L and 31.25 μmol/LwasmadefromtwokindsofNi（Ⅱ） compounds（NiCl2o6H2O,NiSO4o6H2O） .HUVECcelllineculturedinvitrowithNi（Ⅱ） concentration,and then the cell proliferation rate was detected by MTT assay after 72 h,TC50 of NiCl2o6H2O and NiSO4o6H2O was measured on HUVEC. The HUVEC was cultured with Ni（Ⅱ） compounds concentration of TC50,and the cell proliferation rate was detected by MTT assay at different cultured time intervals（24 h,48 h,72 h,96 h and 120 h,respectively）;The lactate dehydrogenase（LDH） contents of the medium supernatants and cell lysate was determined with 2,4-dinitrophenylhydrazine assay after cultured for 72 h,then the LDH release rate was calculated and the Ni（Ⅱ） and Ni（Ⅱ） compounds effects was judged;The cell morphology was observed after cultured for 72 h in medium containing NiCl2o6H2O and NiSO4o6H2O by conventional HE staining,respectively;Analyzed the Ni（Ⅱ） -induced Apoptosis in HUVEC by flow cytometry using Annexin V and Propidium Iodide double staining. Results：The Ni（Ⅱ） IC50 of the two compounds NiCl2o6H2O and NiSO4o6H2O on HUVEC（human umbilical vein endothelial） cells were both of 125 μmol/L. Enhanced cytotoxicity of HUVEC was observed as the Ni（Ⅱ） concentration increased;At the fix concentration,the cytotoxic effects of cell activity increased with the increasing incubated time;No significant difference was found between the LDH release rate of HUVEC cells for the two nickel compounds（P0.05）;However,NiSO4o6H2O caused greater cytotoxic effects on the HUVEC morphology and the apoptosis rate than NiCl2o6H2O（P0.01） .（P0.01） . Conclusion：The HUVEC cytotoxicity of Ni（Ⅱ） s is in a time-and concentration-dependent manner,and the cytotoxic effects HUVEC cell morphology and apoptosi
出处
《现代生物医学进展》
CAS
2010年第2期224-228,F0003,共6页
Progress in Modern Biomedicine
基金
国家973项目<面向生物医学应用的纳米材料
器件和系统研究>(2009CB930000)
国家十五攻关项目<先天性心脏病介入治疗器械的研制>(2001BA706B09)
滚动项目<生物陶瓷封堵器>(2004BA706B11)
深圳重大产业技术攻关计划项目<心血管疾病介入治疗生物材料关键技术及新产品研发>(ZD200904300071A)
