低水平镉暴露对破骨细胞分化的影响

doi:10.11843/j.issn.0366-6964.2019.03.022

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (3): 663-669.doi: 10.11843/j.issn.0366-6964.2019.03.022

低水平镉暴露对破骨细胞分化的影响

贺双江^1,2, 宋瑞龙^1,2, 曹莹^1,2, 刘庆羊^1,2, 张闯^1,2, 刘宗平^1,2*

1. 扬州大学兽医学院, 扬州 225009;
2. 江苏高校动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009

收稿日期:2018-08-20 出版日期:2019-03-23 发布日期:2019-03-23
通讯作者: 刘宗平,主要从事动物营养代谢病与中毒病研究,E-mail:liuzongping@yzu.edu.cn
作者简介:贺双江(1993-),男,山东东营人,硕士生,主要从事动物营养代谢病与中毒病研究,E-mail:1831791326@qq.com
基金资助:
国家重点研发计划支持项目（2016YFD0501208）；国家自然科学基金（31502128；31702304；31702204）；江苏省自然科学基金青年基金（BK20150447）；江苏高校优势学科建设工程资助项目

Effects of Low Level Cadmium Exposure on Osteoclast Differentiation

HE Shuangjiang^1,2, SONG Ruilong^1,2, CAO Ying^1,2, LIU Qingyang^1,2, ZHANG Chuang^1,2, LIU Zongping^1,2*

1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China

Received:2018-08-20 Online:2019-03-23 Published:2019-03-23

摘要/Abstract

摘要：

为研究低水平镉（Cd）暴露对破骨细胞（osteoclast，OC）分化的影响，试验以RAW264.7细胞（单核巨噬细胞系）为材料，在巨噬细胞集落刺激因子（M-CSF）和核因子κB受体活化因子配体（RANKL）存在的条件下，用不同浓度Cd处理4 d；利用CCK-8法检测破骨细胞及其前体细胞活性变化，抗酒石酸酸性磷酸酶（TRAP）染色试验观察破骨细胞生成，激光共聚焦显微镜观察破骨细胞形态变化，蛋白免疫印迹（Western blot）技术和荧光定量聚合酶链式反应（qRT-PCR）检测破骨细胞标志性蛋白及其mRNA水平。结果显示，随着Cd浓度升高，细胞活力受到明显的抑制（P<0.01），并呈浓度-效应关系；与对照组相比，破骨细胞产生的数目和面积均显著或极显著下降（P<0.05或P<0.01）；2、5 μmol·L^-1 Cd处理组破骨细胞封闭带的形成均受到抑制；2和5 μmol·L^-1 Cd处理组破骨细胞特异性蛋白及其mRNA表达量均显著或极显著下降（P<0.05或P<0.01），并呈剂量依赖性。结果表明，低微摩尔水平镉暴露能够抑制破骨细胞的分化。

Abstract:

In order to study the effect of low level cadmium (Cd) exposure on osteoclast (OC) differentiation, RAW264.7 cell (mononuclear macrophage lineage) was used as materials. In the presence of macrophage colony stimulating factor (M-CSF) and receptor-activated nuclear factor κB ligand (RANKL), treatment of different concentrations of Cd were conducted for 4 days. CCK-8 assay was used to detect changes in the viability of osteoclasts and their precursor cells. Tartrate-resistant acid phosphatase (TRAP) staining assay was used to observe the osteoclastogenesis. Laser scanning confocal microscopy was used to observe morphological changes of osteoclasts. The expression of osteoclast marker proteins and mRNA expression were detected by Western blot and qRT-PCR. The results showed that with the increase of Cd concentration, cell viability was significantly inhibited (P<0.01),which showed a concentration-effect relationship. Compared with the control group, the number and area of osteoclast production were significantly or extremely significantly decreased (P<0.05 or P<0.01). In the 2 and 5 μmol·L^-1 Cd treatment groups, the formation of osteoclasts' sealing zone was inhibited. The osteoclast-specific protein and mRNA expression levels of 2 and 5 μmol·L^-1 Cd groups were significantly or extremely significantly decreased (P<0.05 or P<0.01), which showed the dose effect. The above results indicate that low micromolar cadmium exposure can inhibit the differentiation of osteoclasts.

中图分类号:

S856.9

贺双江, 宋瑞龙, 曹莹, 刘庆羊, 张闯, 刘宗平. 低水平镉暴露对破骨细胞分化的影响[J]. 畜牧兽医学报, 2019, 50(3): 663-669.

HE Shuangjiang, SONG Ruilong, CAO Ying, LIU Qingyang, ZHANG Chuang, LIU Zongping. Effects of Low Level Cadmium Exposure on Osteoclast Differentiation[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(3): 663-669.

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低水平镉暴露对破骨细胞分化的影响

Effects of Low Level Cadmium Exposure on Osteoclast Differentiation

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