百色市科学研究与技术发展计划 （20224128） 为这项工作提供了支持。作者感谢伦敦帝国理工学院的 Dev Sooranna 博士和 YMUN 编辑手稿。YYX 和 ZYW 对这项研究的贡献相同。

小药丸对小鼠绝经后骨质疏松症的治疗作用

作者声明不存在利益冲突。

据统计，骨质疏松症在美国每年导致 150 万例骨折，其中绝大多数发生在绝经后妇女身上 27。随着人口老龄化的增加，预计全球未来大部分髋部骨折将发生在亚洲，到 2050 年，全球髋部骨折总数将达到 820 万例28。预防骨折的成本几乎等于治疗骨折的成本，使用药物不可避免地会产生一些副作用29。因此，迫切需要一种更具成本效益的方法，在?...

世界卫生组织 （WHO） 将骨质疏松症 （OP） 定义为一种以骨量减少和骨组织微结构恶化为特征，导致骨脆性增加，从而增加骨折风险的疾病1。骨质疏松症的临床意义在于它可导致骨折，这与高死亡率、发病率和经济成本有关2。绝经后骨质疏松症 （PMOP） 是由绝经后女性雌激素水平降低引起的，导致破骨细胞活性增加，导致骨质流失和骨微结构破坏。这通常会导致骨质疏松症，对健康产生严重影响3。目前的 PMOP 疗法包括雌激素替代疗法、双膦酸盐和甲状旁腺激素，但它们可能具有不同程度的不良反应、长期依从性不足和高成本4。因此，负担得起的草药对于很大一部分人来说是一种可行的选择。
逍遥丸被列入中国药典5，其中含有八种草药成分，包括柴胡、当归、白芍、白术、茯苓、薄荷草、甘草和新鲜生姜。众所周知，所有这些草药都能有效解肝健脾、养血、调节月经周期，该混合物也用于治疗抑郁症6。然而，逍遥丸在骨质疏松症中的作用尚不清楚。
早期研究表明，炎症会导致骨质流失7 ，并且更年期可能会加速与此过程相关的骨密度下降。此外，骨质疏松症的发展与炎症之间存在密切关系。炎症因子白细胞介素-17 （IL-17） 是由 CD4+ T 淋巴细胞的一个亚群 Th17 细胞分泌的促炎因子。这些细胞与多种慢性炎症有关，它们在类风湿性关节炎骨破坏的发展中起着重要作用8。此外，IL-17 刺激核因子-κ B 配体受体激活剂 （RANKL），RANKL 调节破骨细胞生成，导致比骨形成更大的骨吸收9。IL-17 刺激其他破骨细胞因子的表达，如 TNFα、IL-1、IL-6 和 IL-8。它具有与其他炎症因子协同作用的能力，使其成为重要的炎症效应物10。
研究还表明，逍遥丸与炎症之间存在联系。Shi et al.11 和 Fang et al.12 最近证实，小药丸可以分别降低 IL-6 和 TNF-α 的水平。在另一项代谢相关脂肪性肝炎的研究中，据报道，逍遥丸可以上调丙酸的表达，进而抑制 TNF-α 的表达并发挥抗炎作用13。然而，目前尚不清楚逍遥丸是否可以通过 IL-17 介导炎症反应来调节 PMOM 的发展，这是本研究的目的。
本研究通过网络药理学和生物信息学分析预测了逍遥丸靶点与骨质疏松症相关基因的交集，并分析了蛋白质相互作用、GO 和 KEGG 的交叉基因。基于预测结果，可以观察到 IL-17 信号通路中的关键蛋白 Act1 和 IL-6 的表达14,15，以及骨转换标志物碱性磷酸酶 （ALP） 和 I 型胶原 （COL-1），以观察逍遥丸在 PMOM 小鼠模型中的治疗效果。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1ml Sampler</td>
			<td>Guangxi Beilunhe Medical Industrial Group Co.</td>
			<td>JYQ001</td>
			<td>For anesthesia in mice</td>
		</tr>
		<tr>
			<td>4%polyformaldehyde</td>
			<td>Beijing Solarbio Science &#38; Technology Co.,Ltd.</td>
			<td>P1110</td>
			<td>For tissue fixation</td>
		</tr>
		<tr>
			<td>6-0 absorbable suture (angled needle)</td>
			<td>Shanghai Pudong Jinhuan Medical Supplies Co.</td>
			<td>HZX-06</td>
			<td>For postoperative suturing</td>
		</tr>
		<tr>
			<td>Absorbent cotton ball</td>
			<td>Winner</td>
			<td>MIANQIU-500g</td>
			<td>For sterilization and hemostasis</td>
		</tr>
		<tr>
			<td>Adhesive slides</td>
			<td>Jiangsu Shitai Experimental Equipment Co.</td>
			<td>188105</td>
			<td>For tissue sectioning</td>
		</tr>
		<tr>
			<td>Amobarbital</td>
			<td>Sigma Aldrich (Shanghai) Trading Co.</td>
			<td>A-020-1ML</td>
			<td>For anesthesia in mice</td>
		</tr>
		<tr>
			<td>Bluing Solution</td>
			<td>Beijing Solarbio Science &#38; Technology Co.,Ltd.</td>
			<td>G1866</td>
			<td>Blue coloration of the nuclei of cells after the action of hematoxylin differentiation solution</td>
		</tr>
		<tr>
			<td>C57BL/6 mice</td>
			<td>Beijing Vital River Laboratory Animal Technology Co., Ltd.</td>
			<td>(SCXK 2033-0063)</td>
			<td>For use in animal experiments</td>
		</tr>
		<tr>
			<td>Carbon steel surgical blades</td>
			<td>Premier Medical Equipment Co.</td>
			<td>SP239</td>
			<td>For mouse surgery</td>
		</tr>
		<tr>
			<td>CIKS/TRAF3IP2 Rabbit pAb</td>
			<td>BIOSS ANTIBODIES</td>
			<td>bs-6202R</td>
			<td>Binds to Act1 in tissues</td>
		</tr>
		<tr>
			<td>Cole&#39;s Hematoxylin Solution (For Conventional Stain)</td>
			<td>Beijing Solarbio Science &#38; Technology Co.,Ltd.</td>
			<td>G1140</td>
			<td>For staining paraffin sections</td>
		</tr>
		<tr>
			<td>Collagen Type I Polyclonal antibody</td>
			<td>proteintech</td>
			<td>14695-1-AP</td>
			<td>Binds to COL-1 in tissues</td>
		</tr>
		<tr>
			<td>DAB Substrate kit,20x</td>
			<td>Beijing Solarbio Science &#38; Technology Co.,Ltd.</td>
			<td>DA1010</td>
			<td>For tissue color development</td>
		</tr>
		<tr>
			<td>Disposable surgical sheet 50*60CM</td>
			<td>Nanchang Xuhui Medical Equipment Co.</td>
			<td>SP4529777</td>
			<td>For mouse surgery</td>
		</tr>
		<tr>
			<td>EDTA decalcification solution (pH 7.2)</td>
			<td>Beijing Solarbio Science &#38; Technology Co.,Ltd.</td>
			<td>E1171-500ml</td>
			<td>For tissue decalcification</td>
		</tr>
		<tr>
			<td>Enhanced Endogenous Peroxidase Bloching Buffer</td>
			<td>Beyotime Biotechnology</td>
			<td>P0100B</td>
			<td>Sequestration of tissue or cellular endogenous peroxidases</td>
		</tr>
		<tr>
			<td>Environmentally friendly dewaxing clear liquid</td>
			<td>Servicebio</td>
			<td>G1128-1L</td>
			<td>For dewaxing paraffin sections</td>
		</tr>
		<tr>
			<td>Ethyl Alcohol</td>
			<td>CHRON CHEMICALS</td>
			<td>64-17-5 (CAS)</td>
			<td>For dehydration of paraffin sections</td>
		</tr>
		<tr>
			<td>General Purpose Antibody Diluent</td>
			<td>Epizyme Biotech</td>
			<td>PS119L</td>
			<td>For antibody dilution</td>
		</tr>
		<tr>
			<td>Hematoxylin Differentiation Solution</td>
			<td>Servicebio</td>
			<td>G1039-500ML</td>
			<td>For differentiation after hematoxylin staining and removal of excessively bound and non-specifically adsorbed dye from tissues</td>
		</tr>
		<tr>
			<td>Hematoxylin Eosin (HE) Staining Kit</td>
			<td>Beijing Solarbio Science &#38; Technology Co.,Ltd.</td>
			<td>G1120-3*100ml</td>
			<td>For tissue staining</td>
		</tr>
		<tr>
			<td>High quality stainless steel surgical knife handle</td>
			<td>Premier Medical Equipment Co.</td>
			<td>SP0088</td>
			<td>For mouse surgery</td>
		</tr>
		<tr>
			<td>HRP-conjugated Affinipure Goat Anti-Rabbit IgG(H+L)</td>
			<td>proteintech</td>
			<td>SA00001-2</td>
			<td>Binds to primary antibody and amplifies signal</td>
		</tr>
		<tr>
			<td>IL-17A Polyclonal antibody</td>
			<td>proteintech</td>
			<td>13082-1-AP</td>
			<td>Binds to IL-17 in tissues</td>
		</tr>
		<tr>
			<td>IL-6 Polyclonal antibody</td>
			<td>proteintech</td>
			<td>21865-1-AP</td>
			<td>Binds to IL-6 in tissues</td>
		</tr>
		<tr>
			<td>Immunohistochemistry pen</td>
			<td>Beijing Zhongshan Jinqiao Biotechnology Co.</td>
			<td>ZLI-9305 (YA0310)</td>
			<td>For drawing circles in immunohistochemistry</td>
		</tr>
		<tr>
			<td>Medical surgical suture Non-absorbent (ball) 5-0 3.5m</td>
			<td>Yangzhou Yuanlikang Medical Equipment Co.</td>
			<td>FHX-5-2</td>
			<td>For postoperative suturing</td>
		</tr>
		<tr>
			<td>Medical Suture Needles Angle Needles 4*10 3/8</td>
			<td>Chaohu Binxiong Medical Equipment Co.</td>
			<td>FHZ612-4</td>
			<td>For postoperative suturing</td>
		</tr>
		<tr>
			<td>Neutral Balsam</td>
			<td>Beijing Solarbio Science &#38; Technology Co.,Ltd.</td>
			<td>G8590</td>
			<td>As a slice sealer</td>
		</tr>
		<tr>
			<td>PBS(1X)</td>
			<td>Shenzhen Mohong Technology Co.,Ltd</td>
			<td>B0015</td>
			<td>Buffer for slide washing and partial solution dilution</td>
		</tr>
		<tr>
			<td>Protein Free Rapid Blocking Buffer (1X)</td>
			<td>Epizyme Biotech</td>
			<td>PS108P</td>
			<td>Avoiding non-specific binding of proteins</td>
		</tr>
		<tr>
			<td>Rabbit Anti-Bone Alkaline Phosphatase antibody</td>
			<td>BIOSS ANTIBODIES</td>
			<td>bs-6292R</td>
			<td>Binds to alkaline phosphatase in tissues</td>
		</tr>
		<tr>
			<td>Saline</td>
			<td>Affiliated Hospital Youjiang Medical University For Nationalities</td>
			<td>LHN500</td>
			<td>For animals by gavage</td>
		</tr>
		<tr>
			<td>Shaving/Electric clippers</td>
			<td>HANGZHOU HUAYUAN PET PRODUCTS CO., LTD.</td>
			<td>DTJ-002</td>
			<td>For shaving mice</td>
		</tr>
		<tr>
			<td>Stainless Steel Medical Needle Holder 14cm Coarse Needle</td>
			<td>Premier Medical Equipment Co.</td>
			<td>SP784</td>
			<td>For mouse surgery</td>
		</tr>
		<tr>
			<td>Stainless Steel Ophthalmic Forceps 10.5cm Curved (No Hook)</td>
			<td>Zhuoyouyue</td>
			<td>YKNWW-10.5</td>
			<td>For mouse surgery</td>
		</tr>
		<tr>
			<td>Stainless Steel Ophthalmic Scissors/Surgical Scissors 10CM Straight Tip</td>
			<td>Premier Medical Equipment Co.</td>
			<td>ZYJD-10-ZJ</td>
			<td>For mouse surgery</td>
		</tr>
		<tr>
			<td>Stainless Steel Tip Gastric Needle 12 Gauge 55mm Elbow</td>
			<td></td>
			<td>GWJ-12-55W</td>
			<td>For use in mice by gavage</td>
		</tr>
		<tr>
			<td>Tris-EDTA Antigen Repair Fluid (50x)</td>
			<td>proteintech</td>
			<td>PR30002</td>
			<td>For antigen repair of paraffin sections</td>
		</tr>
		<tr>
			<td>Wooden dissecting board 25*16cm</td>
			<td></td>
			<td>JP16*24</td>
			<td>For mouse surgery</td>
		</tr>
		<tr>
			<td>Xiaoyao pills</td>
			<td>Jiuzhitang Co.,Ltd.</td>
			<td>YPG-041</td>
			<td>For animal drug delivery</td>
		</tr>
		<tr>
			<td>Others</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>R 4.3.1</td>
			<td></td>
			<td></td>
			<td>Data processing</td>
		</tr>
		<tr>
			<td>Cytoscape3.9.1</td>
			<td>National Resource for Network Biology</td>
			<td></td>
			<td>Building a regulatory network for traditional Chinese medicine</td>
		</tr>
		<tr>
			<td>ImageJ 1.54f</td>
			<td>National Institutes of Health</td>
			<td></td>
			<td>Image processing for immunohistochemistry results</td>
		</tr>
		<tr>
			<td>Adobe Photoshop 24.0.0</td>
			<td>Adobe</td>
			<td></td>
			<td>For image combination</td>
		</tr>
		<tr>
			<td>GraphpadPrism 9.5</td>
			<td>GraphPad Software</td>
			<td></td>
			<td>Statistical analysis of data</td>
		</tr>
		<tr>
			<td>cellsens Dimension</td>
			<td>OLYMPUS</td>
			<td></td>
			<td>For slicing and photographing</td>
		</tr>
		<tr>
			<td>OLYMPUS BX53</td>
			<td>OLYMPUS</td>
			<td></td>
			<td>For HE staining and immunohistochemical section photography</td>
		</tr>
	</tbody>
</table>

studies on the anti-inflammatory effect of xiaoyao pills in the treatment of postmenopausal osteoporosis in mice

骨质疏松症是老年人和绝经后妇女常见的代谢性疾病，早期无明显症状。在这种情况的后期，患者容易发生骨折，这会严重影响他们的健康和生活质量。全球预期寿命的增加使骨质疏松症成为全球关注的问题。逍遥丹之前 用于治疗抑郁症。此外，该药物似乎具有雌激素样活性，这会影响早期成骨细胞特异性标志物 ALP 和骨细胞外基质主要成分 COL-1 的表达。评估小药丸对小鼠绝经后骨质疏松症 （PMOM） 的影响。通过中医系统药理学 （TCMSP） 数据库获取逍遥丸中各草药成分的靶点信息。利用来自 GeneCards 、 OMIM 、 PharmGkb 、 TTD 、 DrugBank 等网站的信息，通过 Cytoscape 和 String 网络构建草药复合物的调控网络，以评估蛋白质相互作用。对小鼠进行卵巢切除，并用高剂量和低剂量的逍遥丸治疗，并将其与对照组进行比较。通过骨组织的免疫细胞化学评估他们的症状。结果表明，逍遥丸具有通过 IL-17 信号通路缓解去卵巢小鼠 PMOM 症状的能力。这种药物有可能成为治疗骨质疏松症的新型治疗剂。

The World Health Organization (WHO) defines osteoporosis (OP) as a disease characterized by a decrease in bone mass and deterioration of the microarchitecture of bone tissue, leading to an increase in bone brittleness and, thus an increased risk of fracture1. The clinical significance of osteoporosis is that it can lead to fractures, which are associated with high mortality, morbidity, and economic costs2. Postmenopausal osteoporosis (PMOP) is caused by a decrease in estrogen levels in women after menopause, which leads to an increase in osteoclast activity, resulting in bone loss and destruction of bone microstructure. This often causes osteoporosis with a serious impact on health3. Current therapies for PMOP include estrogen replacement therapy, bisphosphonates, and parathyroid hormone, but they can have varying degrees of adverse effects, insufficient long-term compliance, and high costs4. Therefore, affordable herbal medicine is a viable alternative for a large proportion of the population.
Xiaoyao pills are included in the Chinese Pharmacopoeia5, and these contain eight herbal components, including Chai Hu, Angelica sinensis, White paeonia lactiflora, Atractylodes macrocephala, Poria cocos, Menthae Herba, licorice, and fresh ginger. All these herbs are known to be effective in detoxifying the liver and strengthening the spleen, nourishing the blood, and regulating the menstrual cycle, and the mixture has also been used for treating depression6. However, the role of Xiaoyao pills in osteoporosis is unclear.
Early studies have suggested that inflammation can lead to bone loss7, and that the decline in bone density associated with this process may be accelerated by menopause. In addition, there is a strong relationship between the development of osteoporosis and inflammation. The inflammatory factor, interleukin-17 (IL-17), is a pro-inflammatory factor secreted by Th17 cells, a subset of CD4+ T lymphocytes. These cells are associated with several chronic inflammatory conditions, and they play an important role in the development of bone destruction in rheumatoid arthritis8. Additionally, IL-17 stimulates nuclear factor-&#954; B ligand receptor activator (RANKL), which regulates osteoclastogenesis, leading to greater bone resorption than bone formation9. IL-17 stimulates the expression of other osteoclastogenic cytokines such as TNF&#945;, IL-1, IL-6, and IL-8. It has the ability to synergize with other inflammatory factors, making it an important inflammatory effector10.
Studies have also shown a&#160;link between Xiaoyao pills and inflammation. Shi et al.11 and Fang et al.12 have recently confirmed that Xiaoyao pills can reduce the levels of IL-6 and TNF-&#945;, respectively. In another study of metabolism-associated steatohepatitis, it was reported that Xiaoyao pills could upregulate the expression of propionic acid, which in turn inhibits the expression of TNF-&#945; and exerts an anti-inflammatory effect13. However, at present, it is not known whether Xiaoyao pills can regulate the development of PMOM by mediating an inflammatory response through IL-17, which was the aim of this study.
This study predicted the intersection of the targets of Xiaoyao pills and osteoporosis-related genes through network pharmacology and bioinformatics analysis and analyzed the intersecting genes for protein interactions, GO, and KEGG. Based on the predicted results, the expression of Act1 and IL-6, which are key proteins in the IL-17 signaling pathway, can be observed14,15, as well as the bone turnover markers alkaline phosphatase (ALP) and collagen type I (COL-1), to observe the therapeutic efficacy of the Xiaoyao pills in the PMOM mice model.

作者聚焦：将传统中医与现代药理学和基因组学相结合以评估小鼠绝经后骨质疏松症 

逍遥丸治疗小鼠绝经后骨质疏松症的抗炎作用研究

逍遥丸的活性成分和作用靶点 通过检索 TCMSP 数据库并根据口服生物利用度 （OB） ≥ 30% 和类药性质 （DL） ≥ 0.18 的标准进行筛选，发现小药丸中存在 125 种活性成分。其中，成分 6、4、9、13、2、6、83 和 2 分别来自白芍、白术、薄荷、柴胡、当归、茯苓、甘草和生姜。一些活性成分如表 1 所示。此外，从 TCMSP 数据库中共获得 879 个有?...

绝经后骨质疏松症已成为一个全球性的公共卫生问题。本研究的目的是探讨中药小药丸对这种情况的治疗效果和相关机制。

Osteoporosis is a common metabolic disease of elderly and postmenopausal women, with no obvious symptoms during its early stages. In the latter stages of ...

studies-on-anti-inflammatory-effect-xiaoyao-pills-treatment

Research

JoVE Journal

Biology

Studies on the Anti-Inflammatory Effect of Xiaoyao Pills in The Treatment of Postmenopausal Osteoporosis in Mice

291 次观看.  Youjiang Medical University for Nationalities. 本研究的范围是中药逍遥丸对绝经后骨质疏松症小鼠的治疗效果。在这项研究中，我们想确认逍遥丸是否可以发挥治疗效果以及具体机制。在这项研究中，我们将传统中医与现代药理学和基因组学以及关节炎生物信息学相结合，以断言我们所看到的分析将向您展示您的同行。也过程与单一药理学或基因组学相比。分析我们的方案更全面，可能为...

视频: 作者聚焦：将传统中医与现代药理学和基因组学相结合以评估小鼠绝经后骨质疏松症 

Osteoporosis is a common metabolic disease of elderly and postmenopausal women, with no obvious symptoms during its early stages. In the latter stages of this condition, the patients are prone to fractures, and this can seriously affect their health and quality of life. The worldwide increase in life expectancy has made osteoporosis a global concern. The Xiaoyao pills were previously 
used in the treatment of depression. In addition, the drug appeared to have estrogen-like activity, which affected the expression of ALP, an early osteoblast-specific marker, and COL-1, a major component of bone extracellular matrix. Xiaoyao pills were assessed for their effects on postmenopausal osteoporosis (PMOM) in mice. The target information of each herbal component of Xiaoyao pills was accessed through the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Information from GeneCards, OMIM, PharmGkb, TTD, DrugBank, and other websites was used to construct the regulatory network of the herbal complex through Cytoscape and String network to assess the protein interactions. Mice were ovariectomized, and treated with high and low doses of Xiaoyao pills and these were compared to controls. Their symptoms were assessed by immunocytochemistry of bone tissues. The results suggested that Xiaoyao pills had the ability to alleviate the symptoms of PMOM in ovariectomized mice through the IL-17 signaling pathway. This drug has the potential to become a novel therapeutic agent for the treatment of osteoporosis.

Postmenopausal osteoporosis has become a global public health problem. The aim of this study is to explore the therapeutic effects and related mechanisms of the traditional Chinese medicine Xiaoyao pills on this condition.