骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞成骨分化潜能的影响

doi:10.11843/j.issn.0366-6964.2022.04.028

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (4): 1280-1288.doi: 10.11843/j.issn.0366-6964.2022.04.028

骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞成骨分化潜能的影响

龙亚丽¹, 田启会^2*

1. 甘肃农业大学动物医学院, 兰州 730070;
2. 甘肃畜牧工程职业技术学院, 武威 733006

收稿日期:2021-07-29 出版日期:2022-04-23 发布日期:2022-04-25
通讯作者: 田启会,主要从事从事兽医临床教学及研究,E-mail:gsxmtqh@163.com
作者简介:龙亚丽(1992-),甘肃陇南人,兽医硕士,主要从事动物生殖生理研究,E-mail:252218024@qq.com
基金资助:
甘肃省自然科学基金项目(20 JR10RA785)；2021年武威市第一批科技计划项目(cBMSCs应用于犬类骨折修复的基础研究)；甘肃省教育厅：优秀研究生“创新之星”项目(2021CXZX-392)

Effects of Total Flavonoids of Drynariae Rhizoma on Osteogenic Differentiation Potential of Canine Bone Marrow Mesenchymal Stem Cells in Hypoxic Environment

LONG Yali¹, TIAN Qihui^2*

1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
2. Gansu Polytechnic College of Animal Husbandry & Engineering, Wuwei 733006, China

Received:2021-07-29 Online:2022-04-23 Published:2022-04-25

摘要/Abstract

摘要： 拟研究骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞(BMSCs)成骨分化潜能的影响。运用骨碎补总黄酮(TFDR)干预低氧浓度(10%)环境中犬BMSCs 4周后诱导成骨分化，倒置显微镜观察茜素红染色BMSCs钙结节形成，比色法检测碱性磷酸酶(ALP)活性水平，流式细胞术检测细胞线粒体膜电位，激光共聚焦显微镜和RT-PCR检测成骨分化关键基因Runx2、Osterix的表达。结果显示：与对照组相比，10%氧浓度抑制了犬BMSCs钙结节形成和ALP活性(P<0.01)，线粒体膜电位降低(P<0.05)，Runx2、Osterix表达降低(P<0.05或P<0.01)；与低氧组相比，TFDR能显著促进低浓度氧中BMSCs钙结节产生和ALP活性(P<0.05)，线粒体膜电位升高(P<0.05)，促进Runx2、Osterix表达升高(P<0.05或P<0.01)。骨碎补总黄酮能够促进低氧浓度下犬BMSCs向成骨方向分化。

关键词: 缺氧环境, 犬骨髓间充质干细胞, 骨碎补总黄酮, 成骨分化

Abstract: This study aims at studying the effects of the total flavonoids of Drynaria Rhizoma (TFDR) on osteogenic differentiation potential of canine bone marrow mesenchymal stem cells (BMSCs) in hypoxic. TFDR was used to intervene canine BMSCs in a low oxygen concentration (10%) environment for 4 weeks to induce osteogenic differentiation. Observation of calcium nodule formation in BMSCs stained with Alizarin Red by an inverted microscope. The alkaline phosphatase (ALP) activity level was detected by colorimetric method. The cell mitochondrial membrane potential was detected by flow cytometry. The expression of Runx2 and Osterix, the key genes for osteogenic differentiation were detected by RT-PCR and confocal laser microscope. Results were as follows: Compared with the control group, 10% oxygen concentration inhibited the formation of calcium nodules and ALP activity in canine BMSCs (P<0.01), Mitochondrial membrane potential level dropped (P<0.05), and the expression of Runx2 and Osterix decreased (P<0.05 or P<0.01). Compared with the hypoxic group, TFDR can significantly promoted the formation of calcium nodules and ALP activity of BMSCs in Hypoxic (P<0.05), the level of cell mitochondrial membrane potential increased (P<0.05), and Runx2 and Osterix expression increased (P<0.05 or P<0.01). TFDR can promote the osteogenic differentiation of canine BMSCs in hypoxic.

Key words: hypoxic environment, canine bone marrow mesenchymal stem cells, total flavonoids of Drynariae Rhizoma, osteogenic differentiation

中图分类号:

S853.74

龙亚丽, 田启会. 骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞成骨分化潜能的影响[J]. 畜牧兽医学报, 2022, 53(4): 1280-1288.

LONG Yali, TIAN Qihui. Effects of Total Flavonoids of Drynariae Rhizoma on Osteogenic Differentiation Potential of Canine Bone Marrow Mesenchymal Stem Cells in Hypoxic Environment[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1280-1288.

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骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞成骨分化潜能的影响

Effects of Total Flavonoids of Drynariae Rhizoma on Osteogenic Differentiation Potential of Canine Bone Marrow Mesenchymal Stem Cells in Hypoxic Environment

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