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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 study the effect of different concentrations of nickel on cultured human umbilical vein endothelial cells (HUVEC). Methods: Ni (Ⅱ) solution containing 1000, 500, 250, 125, 62.5μmol / L and 31.25μmol / L Ni (Ⅱ) was prepared by two nickel compounds (NiCl2 · 6H2O, NiSO4 · 6H2O) (HUVEC) cell lines were treated with TC50 for 72 h. MTT assay was used to determine the cell proliferation activity. The TC50 of NiCl2 · 6H2O and NiSO4 · 6H2O were determined. The HUVEC cell lines were treated with TC50 Ni (Ⅱ) 48h, 72h, 96h, 120h). The content of lactate dehydrogenase (LDH) in the culture supernatant and cell lysate was detected by 2,4-dinitrophenylhydrazine method at 72h, and the activity of Ni Ⅱ), the effect of Ni (Ⅱ) on the release rate of LDH was evaluated. HE staining was used to observe the cell morphology of the two nickel compounds after 72 hours. The apoptosis was detected by Annexin V-FITC / PI double labeling flow cytometry . Results: The TC50 of HUVEC cells was 125μmol / L, and the cytotoxicity increased with the increase of concentration. The effect of the same concentration of Ni (Ⅱ) on the cell activity increased with the increase of the action time. Two nickel compounds (P> 0.05). The effect of NiSO4 · 6H2O on cell morphology and apoptosis rate was significantly greater than that of NiCl2 · 6H2O (P <0.05). CONCLUSION: The effect of Ni (Ⅱ) on the cytotoxicity of HUVEC is time-and concentration-dependent. The effect of NiSO · 46H2O on HUVEC cell morphology and apoptosis is greater than that of NiCl · 26H2O.