| 摘要: | 骨質疏鬆症是一種骨骼疾病，雖然沒有明顯的臨床症狀，但會增加患者骨折、住院，甚至死亡的風險，因此研究骨質疏鬆症的預防和治療就顯得格外重要。目前骨質疏鬆症的臨床治療藥物都因其副作用而有所限制，而過去曾從天然物中發現具有骨重塑潛力活性的成分，值得進一步開發。LM是一種多年生草本植物，在民間被用作抗發炎劑。本研究旨在開發LM以了解其對骨質疏鬆症的影響。首先，以95%乙醇萃取LM (LME)，再以不同濃度之甲醇進行分層萃取，透過全球天然物社會分子網路 (GNPS) 辨識LME分層萃取物的成分組成，其包括異戊烯醇脂質 (prenol lipids)、固醇類 (steroids)、香豆素 (coumarins) 和黃酮類 (flavonoids) 等。為了評估LME分層萃取物的骨重塑活性，體外試驗以NFκ-B配體受體活化劑 (RANKL) 誘導THP-1細胞進行，結果顯示，60%甲醇 (含) 以下之LME分層萃取物沒有細胞毒性；且LME和 60%甲醇分層萃取物 (LME-6) 顯著降低蝕骨細胞之抗酒石酸酸性磷酸? (TRAP) 活性。在體內試驗，60%甲醇 (含) 以下之LME分層萃取物不影響斑馬魚幼魚之存活。為了解LME及其化合物對糖皮質素誘導骨質疏鬆症 (GIOP) 之作用，本研究完成GIOP斑馬魚模式建立，LME能顯著改善GIOP斑馬魚模式之脊椎發育率。LME-6透過高效液相層析儀 (HPLC) 分離、純化，並以核磁共振儀 (NMR) 確定化合物1、2、3、4之結構，而在3之預處理下可顯著增加GIOP斑馬魚模式之脊椎發育率。本篇研究發現LM具骨重塑活性，並可為糖皮質素誘導骨質疏鬆症之預防，提供植物新藥的開發策略。
Osteoporosis is a skeletal disease. Although it has no obvious clinical symptoms, it makes fractures more likely, increasing the patient's risk of hospitalization and even death. Therefore, study on the prevention and treatment of osteoporosis is particularly important. Currently, most clinical treatments for osteoporosis are limited by their side effects. For this reason, there is potential for development in finding ingredients from natural products that have activity on bone remodeling. LM, a perennial herb, is used as an anti-inflammatory agent in folk medicine. This study aims to develop LM to figure out the effect on osteoporosis. First, LM was extracted with 95% ethanol (LME) and fractionated with various concentrations of methanol. The components of the fractions of LME were identified through the Global Natural Molecular Network of Society (GNPS), including prenol lipids, steroids, coumarins and flavonoids, etc. To evaluate the bone remodeling activity of the fractions of LME, in vitro experiments were conducted using NFκ-B ligand receptor activator (RANKL) to induce THP-1 cells. The results showed that 60% methanol and less fractions of LME had no cytotoxicity; and LME and 60% methanol fraction (LME-6) significantly reduced the tartrate-resistant acid phosphatase (TRAP) activity of osteoclasts. In vivo tests, 60% methanol and less fractions of LME did not affect the survival of zebrafish larvae. To understand the effect of LME and its compounds on glucocorticoid-induced osteoporosis (GIOP), this study completed the establishment of the GIOP zebrafish model. LME could significantly improve the vertebra growth rate on the GIOP zebrafish model. LME-6 was separated and purified by high-performance liquid chromatography (HPLC), and the structures of compounds 1, 2, 3, and 4 were determined by nuclear magnetic resonance (NMR). The pretreatment of 3 could significantly increase the vertebra growth rate on GIOP zebrafish model. This study found that LM had bone remodeling activity and can provide a strategy for the development of new plant drugs for the prevention of glucocorticoid-induced osteoporosis. |
