维生素D介导鸡BMSCs和BMMs共培养体系中破骨细胞的形成

doi:10.11843/j.issn.0366-6964.2019.07.021

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (7): 1509-1517.doi: 10.11843/j.issn.0366-6964.2019.07.021

维生素D介导鸡BMSCs和BMMs共培养体系中破骨细胞的形成

顾建红^1,2,3, 闵雯嫣^1,2,3, 周佳桦¹, 赵玉田^1,2,3, 卞建春^1,2,3, 刘学忠^1,2,3, 袁燕^1,2,3, 刘宗平^1,2,3*

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;
3. 江苏省人兽共患病学重点实验室, 扬州 225009

收稿日期:2019-02-28 出版日期:2019-07-23 发布日期:2019-07-23
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:顾建红(1981-),女,江苏如东人,副教授,博士,主要从事动物营养代谢病与中毒病研究,Tel:0514-87979042,E-mail:jhgu@yzu.edu.cn;闵雯嫣(1994-),女,江苏太仓人,硕士生,主要从事动物钙磷代谢紊乱性疾病的研究。二人为同等贡献作者
基金资助:
国家自然科学基金（31872534；31872533；31302154）；江苏省自然科学基金（BK20181452）；江苏高校优势学科建设工程资助项目（PAPD）；扬州大学大学生创新创业训练计划项目/学术科技创新基金项目（x20180609）

Vitamin D Regulates OC Formation in the Co-culture System of Chicken BMSCs and BMMs

GU Jianhong^1,2,3, MIN Wenyan^1,2,3, ZHOU Jiahua¹, ZHAO Yutian^1,2,3, BIAN Jianchun^1,2,3, LIU Xuezhong^1,2,3, YUAN Yan^1,2,3, LIU Zongping^1,2,3*

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;
3. Jiangsu Key Laboratory of Zoonosis, Yangzhou 225009, China

Received:2019-02-28 Online:2019-07-23 Published:2019-07-23

摘要/Abstract

摘要： 为探究1α，25-（OH）₂D₃对鸡骨髓间充质干细胞（BMSCs）和骨髓单核巨噬细胞（BMMs）体外共培养体系中破骨细胞（OC）形成的影响，笔者对1α，25-（OH）₂D₃处理后的细胞进行抗酒石酸酸性磷酸酶染色（TRAP）、骨吸收功能鉴定、OC标志基因mRNA和蛋白表达的检测。结果表明：1α，25-（OH）₂D₃能够上调BMSCs中核因子κB受体活化因子配体（RANKL）的表达，下调骨保护素（OPG）的表达。1α，25-（OH）₂D₃处理共培养细胞5 d，OC数目较对照组明显增多，骨吸收活性较对照组极显著增强，基质金属蛋白酶9（MMP-9）、Ⅱ型碳酸酐酶（CAⅡ）、组织蛋白酶K（CtsK）及TRAP等标志性蛋白的mRNA和蛋白表达极显著高于对照组（P<0.01），其中10^-8 mol·L^-1 1α，25-（OH）₂D₃效果最明显。结果表明，10^-8 mol·L^-1 1α，25-（OH）₂D₃能够介导鸡BMSCs和BMMs共培养体系中OC的形成，该OC具有骨吸收活性，为进一步研究OC在家禽骨骼及钙磷代谢紊乱性疾病中的作用机制奠定基础。

Abstract: This study aimed to investigate the effect of 1α, 25-(OH)₂D₃ on the formation of osteoclast (OC) in the co-culture system of chicken bone marrow mesenchymal stem cells (BMSCs) and bone marrow mononuclear macrophage cells (BMMs). In the study, tartrate-resistant acid phosphatase (TRAP), bone absorption function identification, and OC marker mRNA and protein expression detection were performed on cells treated with 1α, 25-(OH)₂D₃. The results showed that, 1α, 25-(OH)₂D₃ up-regulated the expression of receptor activator for nuclear factor-κB ligand (RANKL) and down-regulated the expression of osteoprotegerin (OPG) of BMSCs. Compared to the control group, the number of OC was increased in the co-culture system at day 5 treated by 1α, 25-(OH)₂D₃. Meanwhile, the bone resorption activity, the mRNA and protein expression of matrix metalloproteinase 9 (MMP-9), carbonic anhydrase Ⅱ (CAⅡ), cathepsin K and TRAP were significantly enhanced (P<0.01) by 1α, 25-(OH)₂D₃. Among them, 10^-8 mol·L^-11α, 25-(OH)₂D₃ had the most obvious effect. The results confirmed that 10^-8 mol·L^-1 1α, 25-(OH)₂D₃ could mediate the formation of OC, which showed bone resorption activity, in the co-culture system of BMSCs and BMMs from chicken. Our study lays a foundation for further research on the mechanism of poultry OC in bone, calcium and phosphorus metabolic diseases.

中图分类号:

S856.5

顾建红, 闵雯嫣, 周佳桦, 赵玉田, 卞建春, 刘学忠, 袁燕, 刘宗平. 维生素D介导鸡BMSCs和BMMs共培养体系中破骨细胞的形成[J]. 畜牧兽医学报, 2019, 50(7): 1509-1517.

GU Jianhong, MIN Wenyan, ZHOU Jiahua, ZHAO Yutian, BIAN Jianchun, LIU Xuezhong, YUAN Yan, LIU Zongping. Vitamin D Regulates OC Formation in the Co-culture System of Chicken BMSCs and BMMs[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(7): 1509-1517.

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维生素D介导鸡BMSCs和BMMs共培养体系中破骨细胞的形成

Vitamin D Regulates OC Formation in the Co-culture System of Chicken BMSCs and BMMs

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