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      In Vivo Screening for Anti-Osteoporotic Fraction from Extract of Herbal Formula Xianlinggubao in Ovariectomized Mice

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          Abstract

          Background and Objectives

          Traditional Chinese Medicine (TCM) Fufang or formula Xianlinggubao (XLGB) is a prescribed TCM drug in China registered for prevention and treatment of osteoporosis. Fufang in TCM is comprised of a group of herbal compounds contributing in group to the treatment efficacy. The present study aims to identify the bioactive fraction(s) in XLGB extract that account(s) dominantly for its osteogenic effects.

          Methods

          The extract of XLGB formula was separated into three fractions using chromatography, i.e., XLGB-A, XLGB-B and XLGB-C. They were administrated to 4-month old ovariectomized (OVX) mice for 6 weeks to determine which bioactive fraction(s) were more effective for preventing OVX-induced bone loss evaluated by microCT, biomechanical testing and biochemical markers. The main peaks of the key fraction were identified using reference compounds isolated from the fraction. In addition, the effects of the composite compounds in XLGB-B on osteoblasts’ proliferation and mineralization were evaluated in UMR 106 cells.

          Results

          XLGB-B with a yield of 13.0% from herbal Fufang XLGB was identified as the most potential one among the three fractions for prevention of OVX-induced bone loss confirmed with bone mass, bone microarchitecture, bone strength and bone turnover markers. Nine compounds in HPLC fingerprint were identified in the XLGB-B fraction, including phenylpropanoids from Herba Epimedii, terpenes from Radix Dipsaci and coumarins from Fructus Psoraleae. In addition, the identified compounds effectively promoted proliferation and/or mineralization of osteoblast-like UMR 106 cells in vitro.

          Conclusion

          XLGB-B with defined phytochemical structures was screened as the key fraction that demonstrated preventive effects on OVX-induced bone loss in mice. The present study laid down a foundation towards a new generation of herbal Fufang characterized with “less herbal materials for achieving equal treatment efficacy” in development strategy of TCM for prevention of OVX-induced osteoporosis.

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          Most cited references 37

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          Bone histomorphometry: standardization of nomenclature, symbols, and units. Report of the ASBMR Histomorphometry Nomenclature Committee.

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            Cardiovascular effects of Danshen.

             Tsung Cheng (2007)
            Danshen is one of the most versatile Chinese herbal drugs that have been used for hundred of years in the treatment of numerous ailments. Because of its properties of improving microcirculation, causing coronary vasodilatation, suppressing the formation of thromboxane, inhibiting platelet adhesion and aggregation, and protecting against myocardial ischemia, it is widely used either alone or in combination with other herbal ingredients for patients with coronary artery disease and other cardiovascular diseases, in both China and other countries including the United States. This article provides an overview of its history, pharmacology, pharmacokinetics, clinical applications, side effects, interactions with Western drugs, and future prospects in the management of cardiovascular diseases.
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              Rehmannia glutinosa: review of botany, chemistry and pharmacology.

               Zheng Jia,  Mao Li,  Ru Zhang (2008)
              Rehmannia glutinosa, a widely used traditional Chinese herb, belongs to the family of Scrophulariaceae, and is taken to nourish Yin and invigorate the kidney in traditional Chinese medicine (TCM) and has a very high medicinal value. In recent decades, a great number of chemical and pharmacological studies have been done on Rehmannia glutinosa. More than 70 compounds including iridoids, saccharides, amino acid, inorganic ions, as well as other trace elements have been found in the herb. Studies show that Rehmannia glutinosa and its active principles possess wide pharmacological actions on the blood system, immune system, endocrine system, cardiovascular system and the nervous system. Currently, the effective monomeric compounds or active parts have been screened for the pharmacological activity of Rehmannia glutinosa and the highest quality scientific data is delivered to support the further application and exploitation for new drug development.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                19 February 2015
                2015
                : 10
                : 2
                Affiliations
                [1 ]Translational Medicine R&D Center, Institute of Biomedical Engineering and Health Tec, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518000, China
                [2 ]Musculoskeletal Research Laboratory, Department of Orthopaedics & Traumatology, The Chinese University of Hong Kong, Hong Kong SAR, China
                [3 ]Institute of Traditional Chinese Medicine & Natural Products, College of Pharmacy, Jinan University, Guangzhou, 510632, China
                Shanghai University of Traditional Chinese Medicine, CHINA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: XLW YD ZHY XSY GZ LQ. Performed the experiments: XLW YXH BSG MCT FJT YD LZZ XHX NW GZ. Analyzed the data: XLW YXH BSG MCT FJT YD ZHY LZZ XHX NW XSY GZ LQ. Contributed reagents/materials/analysis tools: XLW YXH BSG MCT FJT YD ZHY LZZ XHH NW GZ. Wrote the paper: XLW YXH BSG MCT FJT YD ZHY LZZ XHX NW XSY GZ LQ.

                ‡ These authors contributed equally to this work.

                Article
                PONE-D-14-37412
                10.1371/journal.pone.0118184
                4335011
                25695519

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                Page count
                Figures: 3, Tables: 6, Pages: 17
                Product
                Funding
                The work was supported by Ministry of Science and Technology of the People’s Republic of China (2011ZX09201-201-01, Prof. Ling Qin), National Natural Science Foundation of China (81220108028, Prof. Xinsheng Yao) and Ministry of Education of the People's Republic of China (B13038, Prof. Xinsheng Yao). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
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                All relevant data are within the paper and its Supporting Information files.

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