| The Radix Boehmeriae, dry root of Boehmeria nivea (L.) Gaud, has been widely used as a Chinese herb in gynecology. The previous studies showed that Radix Boehmeriae has a variety of pharmacological activities.The present research included two parts, anti-HBV activity screening and the chemical constituents study of the Radix Boehmeriae. It is useful to identify the characteristics components and active components. We can make use of them to establish convenient, fast and accurate qualitative and quantitative methods in order to develop an objective, specific, scientific quality standard for the quality control of the medicinal herb, to ensure its safe and effective clinical use of the Chinese medicine.Firstly, by using the HepG2.2.15 cell model system which was commonly used at home and abroad to explore the anti-hepatitis B virus (HBV) effect of Radix Boehmeriae extract and the methods were mature and reliable. The result showed that HBeAg and HBsAg were not inhibited by five Radix Boehmeriae extract, whereas the HBV-DNA replication was inhibited by three samples, Radix Boehmeriae-water elution, Radix Boehmeriae-water insoluble and Radix Boehmeriae-40% ethanol elution. The SI were over 2, respectively 2.5,7.2,10.6, that meant these parts effective low toxic. The anti-HBV effect of Radix Boehmeriae extract was not caused by its cytotoxicity to cells but inhibition of DNA replication. Guidance by the above results,it is easily to focus on separation of the active ingredient and to establish quality standards for the active ingredient in the future work, and it also lay basic foundation for the further development of anti-HBV drugs.Secondly, in present study, the compounds of Radix Boehmeriae were chemically studied systemically. As aresult,29 compounds were isolated from the ethanol extract of Radix Boehmeriae with modern chromatography methods, which included 7 triterpenes,5 anthraquinone,2 flavones,3 steroidals,2 phenolic compounds,2 sesquiterpene,1 organic acid and 1 inorganic were identified respectively based on spectroscopic and other chemical evidences. Among them, two were new compounds; and another thirteen ones were isolated from the plant firstly.These compounds were identified as follows: Triterpenes: 3β,11α,19α-trihydroxyurs-12-en-28-oic acid(1), hederagenin(2), oleanolic acid(3),ursolic acid(4), pomolic acid (5),3α,19α-dihydroxyurs-12-en-28-oic acid (6)19a-hydroxy-12-en-3-oxo-28-acid(7)Anthraquinone: polydatin (8), emodin-8-O-β-glucopyranoside (9), physcion (10), emodin (11) 1,2-dihydroxy-6-methoxy-3-methylanthracene-9,10-dione-8-O-βglucoside (12)Flavones: catechin(13), epicatechin(14)Sterols: Daucosterol (15),Stigmasterol-3-O-β-D-glucopyranoside (16),β-sitosterol(17)Phenolic compounds:: ethyl3,4-dihydroxybenzoate (18), Ethyl p-hydroxybenzoate(19)Sesquiterpene 3-oxo-α-ionol (C6Epimer) (20,21)Organic Acid: 9,10, 11-Trihydroxy-(12Z)-12-octadecenoic acid(22)Inorganic: potassium nitrate (23)TLC was studied for the discrimination of sample on the basis of above studies. And the HPLC method for determination of the content of pomolic acid in the Radix Boehmeriae was established. The sample was separated on the column of Restek C18 (4.6mm×250mm,5μm), which eluted with methanol and water(0.5% phosphoric acid).The ratio of methanol and water was 70:30 with detected wavelength at 210 nm, flow rate at 1 ml/min, column temperature at25℃. The result showed that method was simple, accurate and could be used for the quality control of the herb. |
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