Wang Ying, Wang Xueru, Wang Kaijin, Qin Weiwei, Li Ning
Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, School of Pharmacy, Anhui Medical University, Hefei 230032, China.
Cells. 2024 Dec 8;13(23):2028. doi: 10.3390/cells13232028.
Postmenopausal osteoporosis (PMOP) is a bone disease characterized by bone thinning and an increased risk of fractures due to estrogen deficiency. Current PMOP therapies often result in adverse side effects. The traditional medicinal plant is commonly used to strengthen bones and support kidney function, but its role in treating PMOP is not well understood. This study aims to investigate the therapeutic effects of the total extract of (Eocc) on PMOP and to explore the underlying mechanisms. The major components of the extract were identified using HPLC. Transcriptomics was employed to predict potential targets. An osteogenic differentiation model of MC3T3-E1 cells was used in vitro. The osteogenic potential of the Eocc was assessed through CCK-8 cell viability assays, alkaline phosphatase (ALP) staining, Alizarin Red staining, Western blotting, and qPCR. MCF-7 and HEK-293 cells were utilized to evaluate the estrogen-like activity of Eocc. Apoptosis rates were detected by flow cytometry. In vivo, a bilateral ovariectomized mouse model of PMOP was used to further validate the in vitro findings through histopathological analysis and WB results. The results demonstrated that the Eocc promoted the proliferation of MC3T3-E1 cells, increased ALP activity, and stimulated the formation of osteogenic mineralized nodules. It also upregulated the expression of osteogenic markers (Runx2, OCN, OPN, and BSP) at both the protein and mRNA levels. The Eocc induced the activation of ERα both in vitro and in vivo, initiating the Src/PI3K/AKT signaling pathway, leading to the phosphorylation of GSK3β and subsequent osteogenesis. The activation of this pathway also stimulated the phosphorylation of mTOR and p70S6K while downregulating cleaved caspase-3 and caspase-9. Additionally, the Eocc reduced apoptosis during osteogenic differentiation and promoted cell proliferation. These findings suggest that the Eocc facilitates osteoblast proliferation and differentiation, improving bone integrity in PMOP mice, and may represent a promising therapeutic candidate for managing PMOP.
绝经后骨质疏松症（PMOP）是一种以骨质变薄和因雌激素缺乏导致骨折风险增加为特征的骨疾病。目前的PMOP治疗方法常常会产生不良副作用。传统药用植物常用于强健骨骼和支持肾功能，但其在治疗PMOP中的作用尚未得到充分了解。本研究旨在探讨[植物名称]总提取物（Eocc）对PMOP的治疗效果，并探索其潜在机制。使用高效液相色谱法（HPLC）鉴定提取物的主要成分。采用转录组学预测潜在靶点。体外使用MC3T3-E1细胞的成骨分化模型。通过CCK-8细胞活力测定、碱性磷酸酶（ALP）染色、茜素红染色、蛋白质印迹法和定量聚合酶链反应（qPCR）评估Eocc的成骨潜力。利用MCF-7和HEK-293细胞评估Eocc的雌激素样活性。通过流式细胞术检测凋亡率。在体内，使用双侧卵巢切除的PMOP小鼠模型，通过组织病理学分析和蛋白质印迹结果进一步验证体外研究结果。结果表明，Eocc促进MC3T3-E1细胞增殖，增加ALP活性，并刺激成骨矿化结节的形成。它还在蛋白质和mRNA水平上调成骨标志物（Runx2、骨钙素（OCN）、骨桥蛋白（OPN）和骨涎蛋白（BSP））的表达。Eocc在体外和体内均诱导雌激素受体α（ERα）的激活，启动Src/磷脂酰肌醇-3激酶（PI3K）/蛋白激酶B（AKT）信号通路，导致糖原合成酶激酶3β（GSK3β）磷酸化及随后的成骨作用。该信号通路的激活还刺激哺乳动物雷帕霉素靶蛋白（mTOR）和核糖体蛋白S6激酶（p70S6K）的磷酸化，同时下调裂解的半胱天冬酶-3（caspase-3）和半胱天冬酶-9。此外，Eocc减少成骨分化过程中的细胞凋亡并促进细胞增殖。这些发现表明，Eocc促进成骨细胞增殖和分化，改善PMOP小鼠的骨完整性，可能是治疗PMOP的一种有前景的候选药物。
