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      A Method of Hepatocyte Extraction Conjugated with HPLC is Established for Screening Potential Active Components inChinese Medicines—Probing Herba Artemisiae Scopariae as an Exemplifying Approach

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          Abstract

          In order to establish an effective and quick method for screening potential bioactive compounds in Traditional Chinese Medicines (TCMs), hepatocytes were employed for extracting either bifendate, a clinical medicine for liver diseases, or chemicals in Herba Artemisiae Scopariae ( A. Scopariae), a commonly used traditional Chinese medicine for remedying liver diseases such as hepatitis induced by viruses, chemicals or alcohol. After hepatocyte extraction the compounds were analyzed by HPLC, therefore this method was referrred to as hepatocyte extraction conjugated with HPLC (HE-HPLC). In the first part of this study, HE-HPLC showed that bifendate was extracted by hepatocytes and detected by HPLC-DAD which indicated the feasibility of this method. Then in the second part of the study, the potential active components in the A. scopariae extract were studied using HE-HPLC. Six chemicals in the A. scopariae extract, which could bind to hepatocytes in vitro, were detected by HPLC-DAD and three were identified as 7-hydroxy-coumarin (7-OH-C), capillartemisin A and 7-methoxy-coumarin, respectively. In vitro assays showed that 7-OH-C protected HL-7702 hepatocytes from H 2O 2 injury. The results indicated that these compounds could be extracted by hepatocytes, could be detected by HPLC and more importantly were bioactive. It is suggested that HE-HPLC is a useful method for screening potent active components in Chinese medicines used to treat liver diseases.

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          Most cited references 15

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          Frequency and pattern of Chinese herbal medicine prescriptions for chronic hepatitis in Taiwan.

          Chinese herbal medicine (CHM) has been commonly used in treating liver diseases in Asian countries. To conduct a large-scale pharmacoepidemiological study and evaluate the frequency and pattern of CHM prescriptions in treating chronic hepatitis. We obtained the database of traditional Chinese medicine outpatient claims from the national health insurance in Taiwan for the whole 2002. Patients with chronic hepatitis were identified by the corresponding diagnosis of International Classification of Disease among claimed visiting files. Corresponding prescription files were analyzed, and association rule were applied to evaluate the co-prescription of CHM in treating chronic hepatitis. Among the 91,080 subjects treated by CHM for chronic hepatitis, the peak age was in the 40 s, followed by 30 s and 50 s. Male/female ratio was 2.07:1. Long-dan-xie-gan-tang and Saliva miltiorrhiza (Dan-shen) were the most commonly prescribed Chinese herbal formula and single herbal drug, respectively. The most common two-drug prescription was Jia-wei-xia-yao-san plus Saliva miltiorrhiza, and the most common three-drug prescription was Jia-wei-xia-yao-san plus Saliva miltiorrhiza and Artemisia capillaries (Yin-chen-hao). This study showed the utilization pattern of Chinese herbal drugs or formulae in treating chronic hepatitis. Further researches and clinical trials are needed to evaluate the efficacy of these Chinese herbs or its ingredients in treating chronic hepatitis.
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            Bifendate treatment attenuates hepatic steatosis in cholesterol/bile salt- and high-fat diet-induced hypercholesterolemia in mice.

            Effects of bifendate, a synthetic intermediate of schisandrin C (a dibenzocyclooctadiene derivative), on liver lipid contents were investigated in experimentally-induced hypercholesterolemia in mice. Hypercholesterolemia was induced by either chronic administration of cholesterol/bile salt or feeding a high-fat diet containing cholesterol and/or bile salt. Hepatic and serum total cholesterol levels were significantly increased (42-268% and 23-124%, respectively) in cholesterol or high-fat diet-treated mice, when compared with control animals receiving vehicle or normal diet. Hepatic triglyceride level was increased (up to 108%), but serum triglyceride level was significantly reduced by 23-63% in hypercholesterolemic mice. Daily administration of bifendate (0.03-1.0 g/kg, i.g.) for 4 days decreased hepatic levels of total cholesterol (9-37%) and triglyceride (10-37%) in hypercholesterolemic mice. Supplementing the high-fat diet with bifendate (0.25%, w/w) caused decreases in hepatic total cholesterol (25-56%) and triglyceride (22-44%) levels following 7 or 14 days of experiment, respectively, when compared with animals fed with high-fat diet not supplemented with bifendate. While fenofibrate treatment decreased both hepatic and serum lipid levels in hypercholesterolemic mice, bifendate treatment did not reduce serum lipid levels. Bifendate and fenofibrate caused an increase (10-41% and 59-98%, respectively) in hepatic index of hypercholesterolemic mice. The results indicate that bifendate treatment can invariably decrease hepatic (but not serum) lipid levels in various mouse models of hypercholesterolemia.
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              Antitumor-activities of coumarin, 7-hydroxy-coumarin and its glucuronide in several human tumor cell lines.

              Coumarin is found in many medicinal plants and therefore also used in phytomedicine for the treatment of venous diseases. The metabolic pathways of coumarin in the human body lead to the intermediate 7-hydroxy-coumarin and consequent glucuronidation in the intestine and liver. The antitumor activities of coumarin (C) and its known metabolite 7-hydroxy-coumarin (7-OH-C) were tested in several human tumor cell lines. C as well as 7-OH-C inhibited cell proliferation of a gastric carcinoma cell line, a colon-carcinoma cell line (Caco-2), a hepatoma-derived cell line (HepG2) and a lymphoblastic cell line (CCRF CEM) in a concentration-dependent way, the IC50-values were 1.59-3.57 mM for C and 0.68-2.69 mM for 7-OH-C. The glucuronide of 7-OH-C was ineffective in this respect.
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                Author and article information

                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                06 February 2012
                February 2012
                : 17
                : 2
                : 1468-1482
                Affiliations
                [1 ]National Standard Laboratory of Pharmacology for Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing 210029, China; Email: blues-xr@ 123456163.com (X.-R.W.); qzhu40@ 123456yahoo.com.cn (Q.Z.)
                [2 ]Jiangsu Key Laboratory for TCM Formulae Research, Nanjing 210046, China; Email: huayongqing@ 123456126.com
                [3 ]School of Chinese Pharmacy, Shengyang Pharmaceutical University, Shenyang 110016, China; Email: umhyu@ 123456yahoo.com.cn
                [4 ]Department of Clinical Pharmacology, Affiliated Nanjing First Hospital of Nanjing Medical University, Nanjing 210006, China
                Author notes
                [* ] Authors to whom correspondence should be addressed; Email: hongmin72@ 123456hotmail.com (M.H.); fanhongwei178@ 123456sina.com (H.-W.F.); Tel.: +86-25-8679-8184 (M.H.); Fax: +86-25-8679-8188 (M.H.); Tel.: +86-25-5227-1448 (H.-W.F.); Fax: +86-25-5227-1444 (H.-W.F.).
                Article
                molecules-17-01468
                10.3390/molecules17021468
                6268900
                22310168
                © 2012 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/3.0/).

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