Effect and Mechanism of Decreasing Intracellular Zn2+ Influx in the Hypoxia Protection of Nucleus Pulposus Cells
- VernacularTitle:细胞内 Zn2＋内流减少在低氧保护髓核细胞中的作用及其机制
- Author:
Xiaofan YIN
1
;
Jun XU
;
Huijie GU
;
Xuhua WU
;
Jianxing LIU
;
Jiong CHEN
;
ZONGYang-ming
Author Information
1. 上海市闵行区中心医院
- Keywords:
Zinc;
Hypoxia;
Nucleus pulposus cells;
Metabolism
- From:
Chinese Journal of Clinical Medicine
2015;(5):607-612
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect and mechanism of intracellular Zn2+ concentration ([Zn2+ ]i) in hypoxia‐induced regulation of metalloproteinases (MMPs) and extracellular matrix (ECM) expression in nucleus pulposus (NP) cells .Methods:NP cells from SD rats received plate culture at first and then three‐dimensional culture with sodium alginate gel .[Zn2+ ]i was assayed by FluoZin‐3 AM staining .Proteoglycan was assayed by Alcian blue staining .Glycosaminoglycan was detected by 1 ,9‐dimethylmethylene blue (DMMB) assay .And real‐time PCR were used to assay the mRNA expression of α1 type II collagen (COL2A1) ,matrix metalloproteinase 13 (MMP‐13) and a disintegrin and metalloproteinase with a thrombospondin motif 5 (ADAMTS‐5) .The expression of ZRT ,IRT‐like protein 8(ZIP8) was assayed by immunohistochemistry and Western blotting .Results:Interleukin (IL)‐1βand ZnCl2 could significantly increase the [Zn2+ ]i of NP cells ,however ,the effect could be inhibited by hypoxia .Hypoxia did significantly attenuate the decrease of proteoglycan ,glycosaminoglycan ,and COL2A1 mRNA ,which was induced by IL‐1βand ZnCl2 treatment ,in sodium alginate three‐dimensional culture . However ,ZnCl2 inhibited the protective effect of hypoxia .Both an intracellular Zn2+chelator and hypoxia could inhibit the increase of MMP‐13 mRNA expression .IL‐1βand ZnCl2 treatment promoted the increase of ZIP8 expression in NP cells ,however ,hypoxia inhibited ZIP8 expression .Conclusions:Hypoxia may regulate the Zn2+ influx in NP cells . Zn2+ mediates the regulation effect of hypoxia on ECM and MMP‐13 .Perhaps the changes of [Zn2+ ]i are involved in the process of intervertebral disc degeneration .
