锌对大鼠生长板软骨细胞ZnT-7表达的影响

Effects of Zinc on the Expression of ZnT-7 in Growth Plate Chondrocytes

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12h。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;5μmol/L(TPEN)组与对照组相比略有增高,而10μmol/L、20μmol/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 chondrucytes. Methods Growth plate chondrocytes were isolated from rib cartilage of Wistar rat. The cells were treated ~dth zinc chelating agent N,N,N＇,N＇-tetrakis （2-pyridylmethyl）cthylene-diamine （TPEN） of different concentration （0,5,10 and 20 μmol/L） for 12 hours. The expression of rat growth plate chondrocytes specificity collagen type Ⅱ was detected by immunohistochemistry. The legalization of ZnT-7 was eheeked 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 levd of ZnT-7 was slightly higher in the cells treated with 5 μmol/L TPEN than the control group,while it was lower in the cells Ireated 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.