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

doi:10.16829/j.slxb.150758

Abstract

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

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

CLC Number:

Q346

XING Baorui, ZHAO Haiping, LI Guangyu, SUN Hongmei, MA Zefang, TAN Zhanqing, HU Xiao, LI Chenying, WU Jianhua, LIU Zhen. Effect of steroid hormones on osteoprotegerin expression in antler mesenchymal stem cells of the sika deer[J]. ACTA THERIOLOGICA SINICA, 2023, 43(4): 443-449.

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.

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