甾体类激素对梅花鹿鹿茸间充质干细胞骨保护素表达的影响

邢宝瑞; 赵海平; 李光玉; 孙红梅; 马泽芳; 谭展清; 胡肖; 李晨莹; 吴建华; 刘振

doi:10.16829/j.slxb.150758

兽类学报 >

2023 , Vol. 43 >Issue 4: 443 - 449

DOI: https://doi.org/10.16829/j.slxb.150758

研究论文

甾体类激素对梅花鹿鹿茸间充质干细胞骨保护素表达的影响

邢宝瑞 ,
赵海平 ,
李光玉 ,
孙红梅 ,
马泽芳 ,
谭展清 ,
胡肖 ,
李晨莹 ,
吴建华 ,
刘振

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1 青岛农业大学动物科技学院, 青岛 266109;
2 中国农业科学院特产研究所, 长春 130022;
3 山东省安丘市人民医院神经外二科, 潍坊 262100

邢宝瑞(1997-)，男，硕士研究生，主要从事梅花鹿与鹿茸干细胞成骨机制研究.

收稿日期: 2022-11-22

修回日期: 2023-04-12

网络出版日期: 2023-08-02

基金资助

青岛农业大学高层次人才引进项目(1121009)；山东省现代农业产业技术体系特种经济动物产业技术体系(SDAIT-21-01)

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Effect of steroid hormones on osteoprotegerin expression in antler mesenchymal stem cells of the sika deer

XING Baorui ,
ZHAO Haiping ,
LI Guangyu ,
SUN Hongmei ,
MA Zefang ,
TAN Zhanqing ,
HU Xiao ,
LI Chenying ,
WU Jianhua ,
LIU Zhen

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1 Qingdao Agricultural University, Qingdao 266109, China;
2 Institute of Special Animal and Plant Science of Chinese Academy of Agricultural Science, Changchun 130022, China;
3 Department of Neurosurgery, Anqiu People's Hospital, Weifang 262100, China

Received date: 2022-11-22

Revised date: 2023-04-12

Online published: 2023-08-02

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摘要

骨保护素(Osteoprotegerin,OPG)表达量的上调可通过抑制骨质流失的作用来改善骨骼的动态平衡，在前期OPG的表达检测过程中，OPG在鹿茸间充质干细胞(Antler mesenchymal stem cells,RMCs)成骨过程中呈上升趋势，但在RMCs成骨后较对照诱导组并没有显著差异。由于梅花鹿外周血甾体类激素水平[雌二醇(Estradiol,E₂)、甲状旁腺素1-34(Parathyroid hormone 1-34,PTH 1-34)]与鹿茸骨化密切相关，尚不清楚RMCs成骨的分子机制，据此推测，RMCs表达的OPG可能受甾体类激素的调控。本研究旨在探索甾体类激素(E₂和PTH 1-34)对OPG表达水平的影响。本研究采用CCK-8细胞增殖实验、qRT-PCR、Western Blot、成骨诱导、茜素红染色等方法探索了甾体类激素对RMCs增殖、OPG表达和成骨的影响。结果表明，RMCs在成骨分化过程中，PTH 1-34对RMCs增殖的影响无显著差异，可显著促进OPG蛋白的表达水平抑制破骨，通过上调成骨转录因子SP7和BGN的mRNA表达来促进成骨；E₂显著抑制RMCs的增殖，可通过下调OPG蛋白和SP7基因的表达来抑制RMCs成骨。综上，PTH 1-34可上调OPG蛋白的表达，对RMCs成骨具有促进作用；E₂可下调OPG蛋白的表达，对RMCs成骨具有抑制作用。

关键词： 甾体类激素; 鹿茸; 间充质干细胞; 骨保护素; 成骨诱导

本文引用格式

邢宝瑞 , 赵海平 , 李光玉 , 孙红梅 , 马泽芳 , 谭展清 , 胡肖 , 李晨莹 , 吴建华 , 刘振 . 甾体类激素对梅花鹿鹿茸间充质干细胞骨保护素表达的影响[J]. 兽类学报, 2023 , 43(4) : 443 -449 . DOI: 10.16829/j.slxb.150758

Abstract

The upregulation of Osteoprotegerin (OPG) expression could improve the dynamic bone homeostasis by inhibiting bone loss. During the pre-OPG expression assay, OPG showed an upward trend during osteogenesis of Antler mesenchymal stem cells (RMCs), but there was no significant difference in There was no significant difference between RMCs and control group after osteogenesis. Since peripheral blood levels of steroid hormones (Estradiol, E₂; Parathyroid hormone 1-34, PTH 1-34) are closely related to antler osteogenesis, and the molecular mechanism of osteogenesis in RMCs is unknown, it is speculated that the OPG expressed by RMCs may be regulated by steroid hormones. The aim of this study was to explore the effects of steroid hormones (E₂ and PTH 1-34) on OPG expression levels. The experiment explored the effects of steroid hormones on the proliferation, OPG expression and osteogenesis of RMCs using CCK-8 cell proliferation assay, qRT-PCR, Western Blot, osteogenesis induction, and alizarin red staining. The results showed that there was no significant difference in the effects of PTH 1-34 on the proliferation of RMCs during osteogenic differentiation, which could significantly promote the expression level of OPG protein to inhibit osteolysis and promote osteogenesis by upregulating the mRNA expression of osteogenic transcription factors SP7 and BGN; E₂ significantly inhibited the proliferation of RMCs and could inhibit osteogenesis of RMCs by downregulating the expression of OPG and SP7. In conclusion, PTH 1-34 could upregulate the expression of OPG protein to promote osteogenesis of RMCs; E₂ could downregulate the expression of OPG protein to inhibit the osteogenesis of RMCs.

Key words： Steroid hormone; Deer antler; Mesenchymal stem cells; Osteoprotegerin; Osteogenesis induction

参考文献

An Y, Zhao J F, Nie F F, Wu Y, Xia Y C, Li D. 2019. Parathyroid hormone (PTH) promotes ADSC osteogenesis by regulating SIK2 and Wnt4. Biochemical and Biophysical Research Communications, 516 (2):551-557.
Arceo-Mendoza R M, Camacho P M. 2021. Postmenopausal osteoporosis:latest guidelines. Endocrinology and Metabolism Clinics of North America, 50 (2):167-178.
Bhatnagar A, Kekatpure A L. 2022. Postmenopausal osteoporosis:a literature review. Cureus, 14 (9):e29367.
Bielli A, Freitas-de-Melo A, Genovese P, Villagrán M, Tasende C, Ungerfeld R. 2021. Androgen and oestrogen receptors in the growing antlers velvet of adult and yearling pampas deer (Ozotoceros bezoarticus) males. Anatomia, Histologia, Embryologia, 50 (6):947-955.
Bubenik G A, Reyes E, Schams D, Lobos A, Bartos L, Koerner F. 2002. Effect of antiandrogen cyproterone acetate on the development of the antler cycle in southern Pudu (Pudu puda). The Journal of Experimental Zoology, 292 (4):393-401.
Kim J M, Lin C, Stavre Z, Greenblatt M B, Shim J H. 2018. Osteoblast-osteoclast interactions. Connective Tissue Research, 59 (2):99-107.
Gao Z G. 1999. Study on the relationship between antler growth rate and testosterone and estradiol in the deer. Journal of Economic Zoology, 3 (3):27-30. (in Chinese)
Gaspar-López E, Landete-Castillejos T, Estevez J A, Ceacero F, Gallego L, García A J. 2010. Biometrics, testosterone, cortisol and antler growth cycle in Iberian red deer stags (Cervus elaphus hispanicus). Reproduction in Domestic Animals, 45 (2):243-249.
Guo B, Wang S T, Duan C C, Li D D, Tian X C, Wang Q Y, Yue Z P. 2013. Effects of PTHrP on chondrocytes of sika deer antler. Cell and Tissue Research, 354 (2):451-460.
Guo K T, Wang T C, Luo E J, Leng X Y, Yao B J. 2021. Use of network pharmacology and molecular docking technology to analyze the mechanism of action of velvet antler in the treatment of postmenopausal osteoporosis. Evidence-Based Complementary and Alternative Medicine, 2021:7144529.
Jia J. 2014. Study on the mechanism of estrogen-regulated osteoprotegerin protein expression. Ph. D thesis. Tianjin:Tianjin Medical University. (in Chinese)
Li C Y, Zhao H P, Liu Z, McMahon C. 2014. Deer antler - a novel model for studying organ regeneration in mammals. The International Journal of Biochemistry & Cell Biology, 56:111-122.
Li X F, Cui L K, Chen W H, Fang Y, Shen G B, Li Z, Zhang B B, Wu L G. 2021. QiangGuYin modulates the OPG/RANKL/RANK pathway by increasing secretin levels during treatment of primary type I osteoporosis. Evidence-Based Complementary and Alternative Medicine:ECAM, 2021:7114139.
Lincoln G A, Tyler N J. 1994. Role of gonadal hormones in the regulation of the seasonal antler cycle in female reindeer, Rangifer tarandus. Journal of Reproduction and Fertility, 101 (1):129-138.
Luo F C, Li Z Q, Ling Y. 1999. Determination of estradiol and testosterone in deer antler. Journal of Nanjing Military Medical College, 21 (1):18-19. (in Chinese)
Pereira R J, Duarte J M, Negrão J A. 2005. Seasonal changes in fecal testosterone concentrations and their relationship to the reproductive behavior, antler cycle and grouping patterns in free-ranging male pampas deer (Ozotoceros bezoarticus bezoarticus). Theriogenology, 63 (8):2113-2125.
Remmers S J A, de Wildt B W M, Vis M A M, Spaander E S R, de Vries R B M, Ito K, Hofmann S. 2021. Osteoblast-osteoclast co-cultures:a systematic review and map of available literature. PLoS ONE, 16 (11):e0257724.
Shoback D, Rosen C J, Black D M, Cheung A M, Murad M H, Eastell R. 2020. Pharmacological management of osteoporosis in postmenopausal women:an endocrine society guideline update. The Journal of Clinical Endocrinology & Metabolism, 105 (3):587-594.
Sun H M, Xing X M, Cong B, Li C Y, Yang F H. 2007. Research on in vitro culture technology of antler stem cells. Journal of Economic Zoology, 11 (1):7-9, 14. (in Chinese)
Tao D Y, Zhao J X, Deng G I, Jiao J F. 2015. Relationship between velvet antler ossification and PTH and androgen serum levels in Tarim red deer (Cervus elaphus). Journal of Experimental Zoology. Part A, Ecological Genetics and Physiology, 323 (10):696-703.
Udagawa N, Koide M, Nakamura M, Nakamichi Y, Yamashita T, Uehara S, Kobayashi Y, Furuya Y, Yasuda H, Fukuda C, Tsuda E. 2021. Osteoclast differentiation by RANKL and OPG signaling pathways. Journal of Bone and Mineral Metabolism, 39 (1):19-26.
Wang D T, Berg D, Ba H X, Sun H M, Wang Z, Li C Y. 2019. Deer antler stem cells are a novel type of cells that sustain full regeneration of a mammalian organ-deer antler. Cell Death & Disease, 10(6):443.
Yamaguchi A, Sakamoto K, Minamizato T, Katsube K, Nakanishi S. 2008. Regulation of osteoblast differentiation mediated by BMP, notch, and CCN3/NOV. Japanese Dental Science Review, 44 (1):48-56.
Yao B J, Zhang M, Liu M X, Liu Y X, Hu Y Z, Zhao Y. 2018. Transcriptomic characterization elucidates a signaling network that controls antler growth. Genome, 61 (12):829-841.
Yao Z, Getting S J, Locke I C. 2021. Regulation of TNF-induced osteoclast differentiation. Cells, 11 (1):132.
Yasuda H. 2021. Discovery of the RANKL/RANK/OPG system. Journal of Bone and Mineral Metabolism, 39 (1):2-11.
Zhao H P, Cui X Z, Zhang W, Chen G X, Yang W Y, Wang Q W, Li C Y. 2018. Effects of antiandrogen and progesterone on the growth and development of second-crop antlers in six-year-old Merganser deer. Chinese Journal of Traditional Chinese Medicine, 43 (13):2713-2719. (in Chinese)
孙红梅, 邢秀梅, 丛波, 李春义, 杨福合. 2007. 鹿茸干细胞体外培养技术的研究. 经济动物学报, 11 (1):7-9, 14.
罗方成, 李志强, 凌云. 1999. 鹿茸中雌二醇和睾酮含量的测定. 南京军医学院学报, 21 (1):18-19.
赵海平, 崔学哲, 张伟, 陈广信, 杨万云, 王权威, 李春义. 2018. 抗雄性激素和孕酮对 6 岁龄梅花鹿二茬茸生长发育的影响. 中国中药杂志, 43 (13):2713-2719.
高志光. 1999. 梅花鹿鹿茸生长速度与睾酮、雌二醇关系的研究. 经济动物学报, 3 (3):27-30.
贾军. 2014. 雌激素调控骨保护素蛋白表达机制的研究. 天津:天津医科大学博士学位论文.

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