Ethanol extract of Cyathulae Radix inhibits osteoclast differentiation and bone loss

IF 4 2区 医学 Q1 INTEGRATIVE & COMPLEMENTARY MEDICINE
Liying SHI , Liuyi REN , Jinping LI , Xin LIU , Jingjing LU , Lujuan JIA , Baoping XIE , Siyuan TANG , Wei LIU , Jie ZHANG
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引用次数: 0

Abstract

Cyathulae Radix, a traditional Chinese medicine and a common vegetable, boasts a history spanning millennia. It enhances bone density, boosts metabolism, and effectively alleviates osteoporosis-induced pain. Despite its historical use, the molecular mechanisms behind Cyathulae Radix's impact on osteoporosis remain unexplored. In this study, we investigated the effects and mechanisms of Cyathulae Radix ethanol extract (CEE) in inhibiting osteoporosis and osteoclastogenesis. Eight-week-old female mice underwent ovariectomy and were treated with CEE for eight weeks. Micro-computed tomography (micro-CT) assessed histomorphometric parameters, bone tissue staining observed distal femur histomorphology, and three-point bending tests evaluated tibia mechanical properties. Enzyme-linked immunosorbent assay (ELISA) measured serum estradiol (E2), receptor activator for nuclear factor B ligand (RANKL), and osteoprotegerin (OPG) levels. Osteoclastogenesis-related markers were analyzed via Western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, CEE effects on RANKL-induced osteoclast formation and bone resorption were investigated in vitro using tartrate-resistant acid phosphatase (TRAP) staining, qRT-PCR, and WB assay. Compared with the ovariectomy (OVX) group, CEE treatment enhanced trabecular bone density, maximal load-bearing capacity, and various histomorphometric parameters. Serum E2 and OPG levels significantly increased, while Receptor activator of nuclear factor-κB (RANK) decreased in the CEE group. CEE downregulated matrix metallopeptidase 9 (MMP-9), Cathepsin K (CTSK), and TRAP gene and protein expression. In bone marrow macrophages (BMMs), CEE reduced mature osteoclasts, bone resorption pit areas, and MMP-9, CTSK, and TRAP expression during osteoclast differentiation. Compared with DMSO treatment, CEE markedly inhibited RANK, TNF receptor associated factor 6 (TRAF6), Proto-oncogene c-Fos (c-Fos), Nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) expressions, and Extracellular regulated protein kinases (ERK), c-Jun N-terminal kinase (JNK), NF-kappa B-p65 (p65) phosphorylation in osteoclasts. In conclusion, CEE significantly inhibits OVX-induced osteoporosis and RANKL-induced osteoclastogenesis, potentially through modulating the Estrogen Receptor (ER)/RANK/NFATc1 signaling pathway.

蛇床子乙醇提取物可抑制破骨细胞分化和骨质流失
川芎是一种传统中药,也是一种常见的蔬菜,具有上千年的历史。它能增强骨密度,促进新陈代谢,有效缓解骨质疏松症引起的疼痛。尽管 Cyathulae Radix 的使用历史悠久,但其对骨质疏松症产生影响的分子机制仍有待探索。在这项研究中,我们探讨了蛇床子乙醇提取物(CEE)在抑制骨质疏松症和破骨细胞生成方面的作用和机制。八周大的雌性小鼠被切除卵巢,并接受为期八周的 CEE 治疗。显微计算机断层扫描(micro-CT)评估组织形态参数,骨组织染色观察股骨远端组织形态,三点弯曲试验评估胫骨机械性能。酶联免疫吸附试验(ELISA)测定了血清雌二醇(E2)、核因子B配体受体激活剂(RANKL)和骨保护素(OPG)的水平。通过 Western 印迹(WB)和定量实时聚合酶链反应(qRT-PCR)分析了破骨细胞生成相关标记物。此外,还使用耐酒石酸磷酸酶(TRAP)染色、qRT-PCR和WB检测法在体外研究了CEE对RANKL诱导的破骨细胞形成和骨吸收的影响。与卵巢切除(OVX)组相比,CEE治疗提高了骨小梁密度、最大承重能力和各种组织形态学参数。CEE组的血清E2和OPG水平明显升高,而核因子κB受体激活剂(RANK)水平下降。CEE 下调了基质金属肽酶 9(MMP-9)、酪蛋白酶 K(CTSK)和 TRAP 基因和蛋白的表达。在骨髓巨噬细胞(BMMs)中,CEE减少了成熟破骨细胞、骨吸收坑面积以及破骨细胞分化过程中MMP-9、CTSK和TRAP的表达。与 DMSO 处理相比,CEE 能明显抑制破骨细胞中 RANK、TNF 受体相关因子 6(TRAF6)、原癌基因 c-Fos(c-Fos)、活化 T 细胞胞浆核因子 1(NFATc1)的表达,以及细胞外调节蛋白激酶(ERK)、c-Jun N 端激酶(JNK)、NF-kappa B-p65(p65)的磷酸化。总之,CEE能明显抑制OVX诱导的骨质疏松症和RANKL诱导的破骨细胞生成,这可能是通过调节雌激素受体(ER)/RANK/NFATc1信号通路实现的。
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来源期刊
Chinese Journal of Natural Medicines
Chinese Journal of Natural Medicines INTEGRATIVE & COMPLEMENTARY MEDICINE-PHARMACOLOGY & PHARMACY
CiteScore
7.50
自引率
4.30%
发文量
2235
期刊介绍: The Chinese Journal of Natural Medicines (CJNM), founded and sponsored in May 2003 by China Pharmaceutical University and the Chinese Pharmaceutical Association, is devoted to communication among pharmaceutical and medical scientists interested in the advancement of Traditional Chinese Medicines (TCM). CJNM publishes articles relating to a broad spectrum of bioactive natural products, leading compounds and medicines derived from Traditional Chinese Medicines (TCM). Topics covered by the journal are: Resources of Traditional Chinese Medicines; Interaction and complexity of prescription; Natural Products Chemistry (including structure modification, semi-and total synthesis, bio-transformation); Pharmacology of natural products and prescription (including pharmacokinetics and toxicology); Pharmaceutics and Analytical Methods of natural products.
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