Pathomwichaiwat Thanika, Ochareon Pannee, Soonthornchareonnon Noppamas, Ali Zulfiqar, Khan Ikhlas A, Prathanturarug Sompop
Department of Pharmaceutical Botany, Faculty of Pharmacy, Mahidol University, 447 Sri-Ayuthaya Road, Rajathevi, Bangkok 10400, Thailand.
Department of Anatomy, Faculty of Dentistry, Mahidol University, 6 Yothi Road, Rajathevi, Bangkok 10400, Thailand.
J Ethnopharmacol. 2015 Feb 3;160:52-60. doi: 10.1016/j.jep.2014.11.026. Epub 2014 Nov 24.
The stem of Cissus quadrangularis L. (CQ) is used in traditional medicine to treat bone fractures and swelling. Anti-osteoporotic activity of CQ hexane extract has been reported, but the active compounds in this extract remain unknown. Thus, we aimed to identify the active compounds in CQ hexane extract using bioassay-guided isolation.
The CQ hexane extract was fractionated sequentially with benzene, dichloromethane, ethyl acetate, and methanol. The examination of CQ extract and its fractions was guided by bioassays for alkaline phosphatase (ALP) activity during the differentiation of MC3T3-E1 osteoblastic cells. The cells were treated with or without the CQ extract and its fractions for a period of time, and then the stimulatory effect of the alkaline phosphatase enzyme, a bone differentiation marker, was investigated. The compounds obtained were structurally elucidated using spectroscopic techniques and re-evaluated for activity during bone differentiation.
A total of 29 compounds were isolated, viz., triterpenes, fatty acid methyl esters, glycerolipids, steroids, phytols, and cerebrosides. Four new dammarane-type triterpenes were isolated for the first time from nature, and this report is the first to identify this group of compounds from the Vitaceae family. Seven compounds, viz., glycerolipids and squalene, stimulated ALP activity at a dose of 10μg/mL. Moreover, the synergistic effect of these compounds on bone formation was demonstrated.
This report describes, for the first time, the isolation of active compounds from CQ hexane extract; these active compounds will be useful for the quality control of extracts from this plant used to treat osteoporosis.
四角金虎尾（CQ）的茎在传统医学中用于治疗骨折和肿胀。已报道CQ己烷提取物具有抗骨质疏松活性，但该提取物中的活性化合物仍不清楚。因此，我们旨在通过生物测定导向分离法鉴定CQ己烷提取物中的活性化合物。
CQ己烷提取物依次用苯、二氯甲烷、乙酸乙酯和甲醇进行分离。在MC3T3-E1成骨细胞分化过程中，通过碱性磷酸酶（ALP）活性的生物测定来指导对CQ提取物及其馏分的检测。将细胞用CQ提取物及其馏分处理或不处理一段时间，然后研究骨分化标志物碱性磷酸酶的刺激作用。使用光谱技术对获得的化合物进行结构解析，并在骨分化过程中重新评估其活性。
共分离出29种化合物，即三萜类、脂肪酸甲酯、甘油脂、甾体、叶绿醇和脑苷脂。首次从自然界中分离出四种新的达玛烷型三萜，本报告也是首次从葡萄科中鉴定出这一组化合物。七种化合物，即甘油脂和角鲨烯，在10μg/mL的剂量下刺激ALP活性。此外，还证明了这些化合物对骨形成的协同作用。
本报告首次描述了从CQ己烷提取物中分离出活性化合物；这些活性化合物将有助于对用于治疗骨质疏松症的该植物提取物进行质量控制。
