Decreased Zn2+ Influx Underlies the Protective Role of Hypoxia in Rat Nucleus Pulposus Cells
Decreased Zn2+ Influx Underlies the Protective Role of Hypoxia in Rat Nucleus Pulposus Cells
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Zn2 流入减少是缺氧对大鼠髓核细胞的保护作用的基础
DOI：
10.1007/s12011-015-0335-2
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发表时间：
2015-04
影响因子：
3.9
通讯作者：
Dong Jian
中科院分区：
文献类型：
--
作者：
Gu Hui-Jie;Wu Xu-Hua;Li Xi-Lei;Dong Jian
Zn2+是金属蛋白酶的重要组成部分，是金属蛋白酶在软骨中的活性所必需的；然而，Zn2+对髓核（NP）细胞的影响尚未被广泛研究。本文的目的是研究细胞内Zn2+浓度（[Zn2+]i）在缺氧诱导下对NP细胞金属蛋白酶（MMPs）和细胞外基质（ECM）产生的调节作用。将SD大鼠NP细胞培养成单层或海藻酸盐微球。[Zn2+]i采用氟ozin -3 AM染色法测定。采用阿利新蓝染色、免疫化学、1,9-二甲基亚甲基蓝（DMMB）测定和实时荧光定量PCR检测II型胶原蛋白、蛋白多糖、COL2A1、MMP-13和ADAMTS-5 mRNA表达。采用免疫化学和Western blotting检测软骨细胞中主要的Zn2+转运蛋白ZIP8。缺氧可显著抑制白细胞介素(IL)-1β-和zncl2诱导的[Zn2+]i升高。缺氧不能逆转IL-1β和ZnCl2在单层培养中引起的ECM表达下降，但显著减弱IL-1β和ZnCl2处理后藻酸珠培养中蛋白聚糖、糖胺聚糖（GAG）和COL2A1 mRNA表达的下降。然而，ZnCl2抑制了缺氧的保护作用。细胞内Zn2+螯合剂和缺氧均可阻止mmp - 13mrna表达的增加。IL-1β和ZnCl2处理可提高NP细胞中ZIP8的表达，而缺氧可抑制ZIP8的表达。综上所述，缺氧对ECM和MMP-13表达的保护作用与Zn2+内流减少有关。因此，细胞内Zn2+浓度的变化可能与椎间盘退变有关。
Zn2+ is an essential component of metalloproteinases, and is required for their activity in cartilage; however, the effect of Zn2+ on nucleus pulposus (NP) cells has not been widely investigated. The aim of this paper was to investigate the effect of intracellular Zn2+ concentration ([Zn2+]i) in hypoxia-induced regulation of metalloproteinases (MMPs) and extracellular matrix (ECM) production in NP cells. NP cells from Sprague-Dawley (SD) rats were cultured as monolayers or in alginate beads. [Zn2+]i was assayed by FluoZin-3 AM staining. Alcian Blue staining, immunochemistry, 1,9-dimethylmethylene blue (DMMB) assay, and real-time PCR were used to assay collagen II, proteoglycan, and COL2A1, MMP-13, and ADAMTS-5 mRNA expression. ZIP8, a main Zn2+ transporter in chondrocytes, was assayed by immunochemistry and in Western blotting. Interleukin (IL)-1β- and ZnCl2-induced increases of [Zn2+]i were significantly inhibited by hypoxia. Hypoxia did not reverse a decline of ECM expression caused by IL-1β and ZnCl2 in monolayer cultures, but did significantly attenuate the decreases of proteoglycan, glycosaminoglycan (GAG), and COL2A1 mRNA expression following IL-1β and ZnCl2 treatment in alginate bead cultures. However, ZnCl2 inhibited the protective effect of hypoxia. Both an intracellular Zn2+ chelator and hypoxia prevented the increase in MMP-13 mRNA expression. IL-1β and ZnCl2 treatment increased ZIP8 expression in NP cells, and hypoxia inhibited ZIP8 expression. In conclusion, decrease of Zn2+ influx mediates the protective role of hypoxia on ECM and MMP-13 expression. Consequently, changes in intracellular Zn2+ concentration maybe involved in intervertebral disc degeneration.
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影响因子：
4
作者：
Sauer, GR;Smith, DM;Wuthier, RE
通讯作者：
Wuthier, RE
影响因子：
--
作者：
B. Thoms;K. Dudek;J. Lafont;C. L. Murphy
通讯作者：
B. Thoms;K. Dudek;J. Lafont;C. L. Murphy
DOI：
10.1017/cbo9781139047920.006
发表时间：
2012
期刊：
--
影响因子：
--
作者：
D. Cummins
通讯作者：
D. Cummins
影响因子：
8.2
作者：
Gerber, Philipp A.;Bellomo, Elisa A.;Rutter, Guy A.
通讯作者：
Rutter, Guy A.
影响因子：
4.5
作者：
Yang, Shu-Hua;Hu, Ming-Hsiao;Lin, Feng-Huei
通讯作者：
Lin, Feng-Huei