An alternative and simplified approach to identification and test for minimum content of TCM herbal drugs.

Following a decision of the European Pharmacopoeia (Ph. Eur.) Commission, the Traditional Chinese Medicines (TCM) Working Party started a pilot phase to examine the suitability of a high-performance thin-layer chromatography (HPTLC) minimum content test as an alternative to the classical assay in TCM monographs. This approach was evaluated with two TCM herbal drugs: Fritillaria thunbergii bulbs (FTB) and Corydalis rhizome (CYR). Firstly, the existing HPTLC methods were optimised for both drugs. The new methods were applied to the evaluation of multiple samples, and acceptance criteria for the identification, following Ph. Eur. chapter 2.8.25. High-performance thin-layer chromatography of herbal drugs and herbal drug preparations, were set. The HPTLC test for minimum content of markers was then developed and validated. In this test, the intensity of the marker zone in the fingerprint of the sample is compared to the corresponding zone in the reference solution, which has a concentration giving an intensity equivalent to the acceptance criterion. This test gives a pass or fail result rather than a content and can be performed visually (on the images) or by software (using peak profiles from images; PPI). Reproducibility of the HPTLC methods was evaluated in a collaborative trial including six laboratories. In summary, results for FTB from five laboratories were in agreement. The remaining laboratory did not pass the identification of the samples. For CYR, all laboratories presented the same results for identification. In the test for minimum content, one borderline sample passed in four laboratories and failed in two. All laboratories reached similar conclusions for the other seven samples. The HPTLC methods proposed offer a simplified approach to evaluating identity and minimum content of TCM drugs in a single analysis.

Tripterygium glycoside fraction n2: Alleviation of DSS-induced colitis by modulating immune homeostasis in mice.

BACKGROUND: The Tripterygium glycosides (TG) is the main active extractive of Tripterygium wilfordii Hook F and is widely used in clinical practice to treat inflammatory diseases (including inflammatory bowel disease). However, due to its severe toxicity, TG is restricted to the treatment of many diseases. Therefore, it is necessary to study a new method to obtain the attenuated and synergistic extracts from TG. PURPOSE: Tripterygium glycosides-n2 (TG-n2) was obtained from TG by a new preparation method. In this study, we aimed to investigate the difference in the chemical compositions between TG and TG-n2, further explored its toxicity and therapeutic effects on DSS-induced colitis in mice. METHODS: The major chemical compositions of TG and TG-n2 were analyzed by ultra-performance liquid chromatography (UPLC). Subsequently, acute toxicity test was applied to evaluate the toxicity difference between TG and TG-n2. Dextran sulfate sodium (DSS)-induced acute colitis model was used to explore the therapeutic effect of TG and TG-n2 and their potential mechanisms of action. RESULTS: We found that the chemical compositions of TG-n2 is different from TG. The main difference is the ratio of triptriolide (T11) / triptolide (T9). Acute toxicity test proved that TG-n2 was less toxic than TG. Base on this, further studies showed that TG-n2 has a similar therapeutic effect as compared to TG on attenuating the symptoms of colitis, such as diarrhea, bloody stools, body weight loss, colonic atrophy, histopathological changes, inhibiting cytokines secretion and reducing absolute lymph number. In addition, TG and TG-n2 can increase the apoptosis of T lymphocyte in vivo. Further investigated showed that TG and TG-n2 could increase the expressions of Bax and p62 on CD3-positive T cells. CONCLUSION: This study showed that oral administration of TG-n2 is safer than TG. Moreover, the attenuated TG-n2 has the similar therapeutic effect on treating experimental colitis in mice when compared to TG. Its mechanism may be related to activating the expression of Bax in T cells and inducing T cells autophagy to regulate the survival of T lymphocytes in colitis mice, thus reducing inflammation in colon.

Effects of triterpenes from Ganoderma lucidum on protein expression profile of HeLa cells.

To elucidate the cytotoxicity mechanism of Ganoderma triterpenes, a chemoproteomic study using five purified ganoderic acids, ganoderic acid F (GAF), ganoderic acid K (GAK), ganoderic B (GAB), ganoderic acid D (GAD) and ganoderic acid AM1 (GAAM1) was conducted. GAF, GAK, GAB, GAD and GAAM1 treatment for 48 h inhibited the proliferation of HeLa human cervical carcinoma cells with IC(50) values of 19.5+/-0.6 microM, 15.1+/-0.5 microM, 20.3+/-0.4 microM, 17.3+/-0.3 microM, 19.8+/-0.7 microM, respectively. The protein expression profiles of HeLa cells treated with each ganoderic acid at dose of 15 microM for 48 h were checked using two-dimensional electrophoresis (2-DE). The possible target-related proteins of ganoderic acids, i.e. proteins with same change tendency in all five ganoderic acids-treated groups compared with control, were identified using MALDI-TOF MS/MS. Twelve proteins including human interleukin-17E, eukaryotic translation initiation factor 5A (eIF5A), peroxiredoxin 2, ubiquilin 2, Cu/Zn-superoxide dismutase, 14-3-3 beta/alpha, TPM4-ALK fusion oncoprotein type 2, PP2A subunit A PR65-alpha isoform, nucleobindin-1, heterogeneous nuclear ribonucleoprotein K, reticulocalbin 1 and chain A of DJ-1 protein were identified. Ganoderic acids might exert their cytotoxicity by altering proteins involved in cell proliferation and/or cell death, carcinogenosis, oxidative stress, calcium signaling and ER stress.

Microbial transformation of dehydrocostuslactone by Mucor polymorphosporus.

Biotransformation of dehydrocostuslactone (1) by Mucor polymorphosporus yielded four compounds, and their structures were identified as 11alpha,13-dihydrodehydrocostuslactone (2), 3alpha-hydroxy-11alpha,13-dihydrodehydrocostuslactone (3), 3beta-hydroxy-4beta,15,11alpha,13-tetrahydrodehydrocostuslactone (4) and 2beta-hydroxyl-11alpha,13-dihydrodehydrocostuslactone (5), respectively, on the basis of their spectral data. Among them, compound 5 is a new compound.

Antiproliferative activity of the extract of Gleditsia sinensis fruit on human solid tumour cell lines.

BACKGROUND: The fruit extract of Gleditsia sinensis Lam. (GSE) is a traditional herbal medicine that is saponin-rich. However, its activity on solid tumour cell lines has never been demonstrated. METHODS: The activity of GSE was demonstrated in four cancer cell lines (breast cancer MCF-7, MDA-MB231, hepatoblastoma HepG2 and oesophageal squamous carcinoma cell line SLMT-1) using MTT assay, anchorage-independent clonogenicity assay, DNA laddering and in situ cell death detection. RESULTS: The mean MTT(50) (the mean concentration of GSE to reduce MTT activity by 50%) ranged from 16 to 20 microg/ml of GSE. An anchorage-independent clonogenicity assay showed that all of the four solid tumour cell lines gradually lost their regeneration potential after treatment with GSE, DNA fragmentation and TUNEL analysis demonstrated that the action of GSE is both dose- and time course-dependent. CONCLUSIONS: Our results suggest that GSE has a cytotoxic activity and can induce apoptosis in human solid tumour cell lines.

A new phenolic glycoside and a new trans-clerodane diterpene from Conyza blinii.

A new phenolic glycoside, 4-propionyl-2,6-dimethoxyphenyl beta-D-glucopyranoside (1) and a new trans-clerodane diterpene named 19-deacetylconyzalactone (2), were isolated from the aerial parts of Conyza blinii.

[Non-anthraquinone constituents from Rheum sublanceolatum C. Y. Cheng et Kao].

OBJECTIVE: Study on the non-anthraquinone constituents from rhizoma and radix of Rheum sublanceolatum. METHOD: The constituents were isolated through column chromatography and identified on the basis of their physiochemical and spectral data. RESULT: Six non-anthraquinone constituents were isolated and identified as n-octacosanic acid, sitosterol, daucosterol, 2-methyl-5-carboxymethyl-7-hydroxychromone, piceatannol and 6-hydroxymusizin-8-O-beta-D-glucopyranoside. CONCLUSION: All these compounds were firstly isolated from R. sublanceolatum.

[Studies on liposoluble constituents from the aerial parts of Siegesbeckia orientalis L].

Eight compounds were isolated from the aerial parts of Siegesbeckia orientalis L. and their structures were elucidated by spectroscopic methods(IR, EI-MS, 13C-NMR, 1H-NMR, 1H-1H COSY, 1H-1H NOESY and 1H-13C COSY). Compounds I and II are new natural products and named siegesesteric acid(I) and siegesetheric acid(II), their structures were confirmed as ent-17-acetoxy-18-isobutyryloxy-16(alpha)-kauran-19-oic acid(I) and ent-17-ethoxy-16(alpha)-(-)-kauran-19-oic acid(II). The others were identified as known compounds: ent-16 beta, 17-dihydroxy-kauran-19-oic acid (III), kirenol(IV), beta-sitosteryl glucoside(V), heneicosanol(VI), methyl arachidate(VII) and beta-sitosterol(VIII). These compounds, except kirenol and beta-sitosterol, were isolated for the first time from the title plant.

[Chemical components of volatile oil from Echinops grijisii Hance].

The volatile oil obtained from Echinops grijisii roots was analysed by GC-MS method. Twenty four constituents have been identified from the oil, among which cis-beta-farnesene and 5-(3-buten-1-ynyl)bithiophene are the main ones.

[Phytoecdysteroids of Rhaponticum uniflorum root].

Three phytoecdysteroids I, II and III were isolated from the root of Rhaponticum uniflorum (L.) DC. and their structures were elucidated by chemical and spectroscopic methods (UV, IR, EI-MS, FAB-MS, 1HNMR, 13CNMR, 1H-1H COSY, 1H-1H NOESY, 1H-13CCOSY and CD). Compound II is new and named rhapontisterone, its structure was confirmed as (20R, 22R, 24S)-2 beta, 3 beta, 11 alpha, 14 alpha, 20, 22, 24-heptahydroxy-5 beta cholest-7-en-6-one. The other two, I and III, were identified as ecdysterone and turkesterone, respectively, both are known compounds, but turkesterone was isolated for the first time from the title plant.