锌对大鼠生长板软骨细胞ZnT-7表达的影响

摘要

目的 研究锌对大鼠生长板软骨细胞锌转移体-7(ZnT-7)表达的影响及其规律.方法 原代培养Wistar大鼠肋生长板软骨细胞,不同浓度的锌螯合剂TPEN分别处理培养生长板软骨细胞12 h.免疫组化检测生长板软骨细胞特异性Ⅱ型胶原表达.免疫荧光技术对ZnT-7进行定位研究,实时RT-PCR和Western blot检测znT-7表达.结果 Ⅱ型胶原免疫组化检测阳性.ZnT-7在生长板软骨细胞中定位于高尔基体上;Western blot和RT-PCR结果显示;5 μmol/L(TPEN)组与对照组相比略有增高,而10 μmol/L、20 μmol/L组与对照组相比逐渐降低.结论 ZnT-7定位于高尔基体上,同时在缺锌的条件下其对锌离子稳定有一定的代偿作用.%Objective To study the effects of zinc on the expression of zinc transporter-7( ZnT-7) in the proliferation of the rat growth plate chondrocytes. Methods Growth plate chondrocytes were isolated from rih cartilage of Wistar rat. The cell3 were treated with zinc chelating agent N, N, N', N'-tetrakis (2-pyridylmethyl )ethylene-diamine (TPEN) of different concentration (0,5,10 and 20 μmol/L) for 12 horns. The expression of rat growth plate chondrocytes specificity collagen type Ⅱ was detected by immunohisloehemistiy. The localization of ZnT-7 was checked by immunofluorescent staining. Real-time RT-PCR and Western blot were used to measure the expression level of ZNT-7 in the cell. Results The result of the immunofluorescence showed that ZnT-7 located in the Golgi apparatus. The expression level of ZnT-7 was slightly higher in the cells treated with 5 μ-mol/L TPEN than the control group, while it was lower in the cells treated with 10 or 20 μmol/L TPEN than the control group. Conclusion ZnT-7 locates in Golgi apparatus and maintains the zinc ion stabilization in the condition of the zinc depletion.