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 H2O2 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.
Highlights
Traditional Chinese medicines (TCMs) are one of the major clinical medical treatments in China and have been demonstrated to be effective by their use in clinical practice for thousands of years.scientists are constantly attempting to determine which chemical(s) contribute to the effects of traditional Chinese medicine (TCM)
Bifendate was detected at 6.3 min in the desorption eluate which indicated that bifendate could bind to hepatocytes. (D) Washing eluate after four washes and eight washes
The results all above showed that (1) bifendate, a clinically effective compound used to treat liver diseases, was detected by hepatocyte extraction conjugated with HPLC (HE-HPLC), (2) at least one compound extracted in A. scopariae by HE-HPLC could protect hepatocytes from injury to a certain extent in vitro
Summary
Traditional Chinese medicines (TCMs) are one of the major clinical medical treatments in China and have been demonstrated to be effective by their use in clinical practice for thousands of years.scientists are constantly attempting to determine which chemical(s) contribute to the effects of TCMs. Traditional Chinese medicines (TCMs) are one of the major clinical medical treatments in China and have been demonstrated to be effective by their use in clinical practice for thousands of years. The extraction and purification of compounds from TCMs is time-consuming and aimless. The screening of bioactive compounds carried out in animal models are laborious and inappropriate for the direct screening of bioactive components in TCMs. Modern pharmacological studies have shown that drug action is usually demonstrated by the drug combining with receptors, enzymes or channels on cell membranes. The interaction of compounds with cells has been successfully used as a basis for the hypothesis of bioactive components in Chinese medicines [1,2,3,4]. In order to identify a quick and specific method to screen bioactive components in a complicated system of TCMs used to prevent liver injury, rat primary hepatocytes, bifendate and
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