葛根素缓解镉对大鼠股骨胫骨中成骨细胞分化的抑制作用

doi:10.11843/j.issn.0366-6964.2022.02.029

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (2): 628-636.doi: 10.11843/j.issn.0366-6964.2022.02.029

葛根素缓解镉对大鼠股骨胫骨中成骨细胞分化的抑制作用

张雪晴^1,2, 仝锡帅^1,2,3, 王果帅^1,2, 卞建春^1,2,3, 刘学忠^1,2, 袁燕^1,2, 邹辉^1,2, 沈小兰⁴, 刘宗平^1,2,3*, 顾建红^1,2*

1. 扬州大学兽医学院, 扬州 225009;
2. 江苏省动物重要疫病与人兽共患病防控协同创新中心, 扬州 225009;
3. 扬州大学农业科技发展研究院(国际联合实验室), 扬州 225009;
4. 兴化市合陈畜牧兽医站, 泰州 225733

收稿日期:2021-05-28 出版日期:2022-02-23 发布日期:2022-03-02
通讯作者: 顾建红,主要从事动物营养代谢病与中毒病研究,E-mail:jhgu@yzu.edu.cn;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:张雪晴(1997-),女,甘肃武威人,硕士生,主要从事动物中毒病及营养代谢病研究,E-mail:xueqingz0224@foxmail.com;仝锡帅(1988-),男,山东曹县人,博士,主要从事动物营养代谢病与中毒病研究,E-mail:xstong@yzu.edu.cn
基金资助:
国家自然科学基金（31872534；31872533；32102732）；江苏省自然科学基金（BK20181452；BK20210806）；江苏高校优势学科建设工程（PAPD）

Puerarin Alleviates the Inhibitory Effect of Cadmium on the Femoral and Tibial Osteoblast Differentiation of Rat

ZHANG Xueqing^1,2, TONG Xishuai^1,2,3, WANG Guoshuai^1,2, BIAN Jianchun^1,2,3, LIU Xuezhong^1,2, YUAN yan^1,2, ZOU Hui^1,2, SHEN Xiaolan⁴, LIU Zongping^1,2,3*, GU Jianhong^1,2*

1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
2. Jiangsu Collaborative Innovation Center for Prevention and Control of Important Animal Diseases and Zoonoses, Yangzhou 225009, China;
3. Institute of Agricultural Science and Technology Development(Joint International Research Laboratory of Agriculture & Agri-Product Safety), Yangzhou University, Yangzhou 225009, China;
4. Hechen Animal Husbandry and Veterinary Station of Xinghua, Taizhou 225733, China

Received:2021-05-28 Online:2022-02-23 Published:2022-03-02

摘要/Abstract

摘要： 旨在探究葛根素对镉（cadmium，Cd）抑制大鼠股骨胫骨中成骨细胞（osteoblast，OB）分化的缓解效应。选取40只3周龄SD雄性大鼠，随机分为4组，分别为对照组（CON）、镉组（Cd）、葛根素组（Pur）和镉+葛根素组（Cd+Pur），每组10只。其中，对照组和镉组大鼠灌服超纯水，连续5周；含葛根素组（葛根素组和镉+葛根素组）大鼠每日灌服葛根素溶液（200 mg·kg^-1BW），连续5周；从第5周开始，对照组与葛根素组大鼠每日腹腔注射生理盐水，含镉组（镉组和镉+葛根素组）大鼠每日腹腔注射醋酸镉（2.5 mg·kg^-1BW），连续1周。通过原子吸收光谱法检测大鼠股骨胫骨中Cd、钙（Ca）与磷（P）的含量，qRT-PCR方法检测OB标志基因（RUNX2、OCN、ALP和OSX）的水平，Western blot检测Wnt/β-catenin信号通路相关蛋白（β-catenin、Wnt3A、LEF1和TCF1）的表达。结果显示，与对照组相比，镉组股骨和胫骨中镉含量极显著上调（P<0.01），OB标志基因均下调，Wnt/β-catenin信号通路相关蛋白极显著下调（P<0.01），但Ca和P含量无显著变化（P>0.05）；葛根素组股骨胫骨中镉、Ca与P含量无显著变化（P>0.05），OB标志基因（RUNX2、ALP和OCN）极显著上调（P<0.01），Wnt/β-catenin信号通路相关蛋白极显著上调（P<0.01）；与镉组相比，镉+葛根素组股骨胫骨中镉含量极显著下调（P<0.01），OB标志基因极显著上调（P<0.01），Wnt/β-catenin信号通路相关蛋白极显著上调（P<0.01），但骨中Ca与P含量无显著变化（P>0.05）。结果表明，葛根素对镉抑制大鼠OB分化具有一定的缓解效应，该过程涉及Wnt/β-catenin信号通路。

关键词: 葛根素, 镉, 成骨细胞分化, Wnt/β-catenin信号通路

Abstract: The aim of this study was to investigate the alleviation effect of puerarin on the differentiation of osteoblasts inhibited by cadmium (Cd) in rat femur and tibia. Forty 3-week-old SD male rats were randomly divided into 4 groups with 10 rats per group:control group (CON), cadmium group (Cd), puerarin group (Pur) and cadmium plus puerarin group (Cd + Pur). The rats in the control group and the cadmium group were gavaged with ultrapure water for 5 consecutive weeks. The rats related to puerarin group (puerarin group and cadmium plus puerarin group) were gavaged with puerarin solution (200 mg·kg^-1BW) daily for 5 consecutive weeks. From the fifth week, rats in control group and puerarin group were intraperitoneally injected with normal saline, and the cadmium containing groups (cadmium group and cadmium plus puerarin group) were intraperitoneally injected with cadmium acetate (2.5 mg·kg^-1BW) daily for 1 consecutive week. The contents of cadmium, calcium (Ca) and phosphorus (P) of the femur and tibia in rats were detected by atomic absorption spectrometry. The level of osteoblastic marker genes (RUNX2, OCN, ALP and OSX) were detected by qRT-PCR. The expression of Wnt/β-catenin signaling pathway-related proteins (β-catenin, Wnt3A, LEF1 and TCF1) were detected by Western blot. The results showed that the content of Cd of the femur and tibia in cadmium group was significantly up-regulated (P<0.01), the level of osteoblastic marker genes were down-regulated, the expression of Wnt/β-catenin signaling pathway-related proteins were extremely significantly down-regulated (P<0.01) compared with the control group, but the content of Ca and P were not significantly changed (P>0.05). The content of Cd, Ca and P of the femur and tibia in puerarin group was not significantly changed (P>0.05), the level of osteoblastic marker genes (RUNX2, ALP and OCN) were extremely significantly up-regulated (P<0.01), and the expression of Wnt/β-catenin signaling pathway-related proteins were extremely significantly up-regulated (P<0.01). The content of Cd of the femur and tibia in the cadmium plus puerarin group was extremely significantly down-regulated (P<0.01), the level of osteoblastic marker gene was extremely significantly up-regulated (P<0.01), and the Wnt/β-catenin signaling pathway-related proteins were extremely significantly up-regulated (P<0.01) compared with the cadmium group, but the content of Ca and P in the femur and tibia had no significant changes (P>0.05). The results showed that puerarin had an alleviation effect on the differentiation of osteoblasts inhibited by cadmium in rats, which was involved in Wnt/β-catenin signaling pathway.

Key words: puerarin, cadmium, osteoblastic differentiation, Wnt/β-catenin signal pathway

中图分类号:

张雪晴, 仝锡帅, 王果帅, 卞建春, 刘学忠, 袁燕, 邹辉, 沈小兰, 刘宗平, 顾建红. 葛根素缓解镉对大鼠股骨胫骨中成骨细胞分化的抑制作用[J]. 畜牧兽医学报, 2022, 53(2): 628-636.

ZHANG Xueqing, TONG Xishuai, WANG Guoshuai, BIAN Jianchun, LIU Xuezhong, YUAN yan, ZOU Hui, SHEN Xiaolan, LIU Zongping, GU Jianhong. Puerarin Alleviates the Inhibitory Effect of Cadmium on the Femoral and Tibial Osteoblast Differentiation of Rat[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 628-636.

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葛根素缓解镉对大鼠股骨胫骨中成骨细胞分化的抑制作用

Puerarin Alleviates the Inhibitory Effect of Cadmium on the Femoral and Tibial Osteoblast Differentiation of Rat

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