[UPLC and HPLC analysis on contents of astilbin and engeletin in dong medicine "sunl gaems" of Guizhou origin by QAMS].

This study aimed to simultaneously determine the contents of astilbin and engeletin in dong medicine "sunl gaems" of Guizhou origin by quantitative analysis of multi-components by single marker (QAMS), with astilbin as the internal standard substance. On UPLC and HPLC chromatograms, different models of instruments were used to investigate relative correction factors (RCF), in order to discuss the interoperability of RCFs established in different chromatographic systems. The engeletin content was calculated based on the established RCFs and compared by the one point external standard method and the external standard working curve method, in order to verify the accuracy of QAMS. According to the result, in different chromatograms, the ratios between RCF and retention time of engeletin and astilbin had a good reproducibility, with RSD between 2.0% and 1.8%, both were less than 3%. The relative differences among results of QAMS, the external standard working curve method of dong medicine "sunl gaems" ranged between 1.6% and 3.9%, with RSD between 2.02%-0.80% in line with relevant requirements and Pearson correlation coefficient at 0.9998 (P <0.01). The findings showed that QAMS was an accurate, reliable and highly reproducible method to determine the contents of astilbin and engeletin in dong medicine "sunl gaems" of Guizhou origin and so could be used to control the inherent quality of the herb.

[Study on scavenging activity to DPPH free radical of different polarity components in Guizhou Miao medicine "bod zangd dak"].

The paper is aimed to search more natural plant antioxidants and further research and develop new medicinal plant resources in Guizhou. The Guizhou special miao medicine "bod zangd dak" was extracted with 60% ethanol. The antioxidant activity of the different polarity components separated from the extract was tested by DPPH method with ascorbic acid as positive control. The results showed that the IC50 of the different polarity components was as following: ascorbic acid (0.033 4 g x L(-1)) < ethyl acetate components (0.052 3 g x L(-1)) < total tannins components (0.054 9 g x L(-1)) < 60% ethanol extraction components (0.076 7 g x L(-1)) < butanol extraction components (0.110 g x L(-1)) < water-soluble polysaccharides components (0.168 g x L(-1)) < water extraction components (0.174 g x L(-1)) < water components after extraction (0.226 g x L(-1)) < total polysaccharides components (0.645 g x L(-1)). It is concluded that the different polarity components have different free radical scavenging activity and that provides a scientific basis for further search of the active ingredients and the activive mechanism.

Apoptosis sensitization by Euphorbia factor L1 in ABCB1-mediated multidrug resistant K562/ADR cells.

In this article, reversal activities of Euphorbia factor L1 (EFL1) against ABCB1-mediated multidrug resistance (MDR) and apoptosis sensitization in K562/ADR cells are reported. EFL1 decreased the IC50 values of anticancer agents in K562/ADR cells over-expressing ABCB1. However, EFL1 did not affect the IC50 values of anticancer agents in sensitive K562 cells. Additionally, EFL1 increased the intracellular accumulation of rhodamine 123 and doxorubicin in K562/ADR cells without affecting their accumulation in K562 cells. Furthermore, EFL1 sensitized the apoptosis triggered by vincristine in K562/ADR cells via mitochondrial pathway, as confirmed by Annexin V-FITC/PI detection and western blot. At the same time, EFL1 did not influence the apoptosis induced by vincristine in K562 cells. Western blot results showed that EFL1 did not affect the phosphorylation level of AKT and ERK in K562 and K562/ADR cells. Finally, EFL1 did not down-regulate protein expression of ABCB1.

[Content detemination of total tannin in Smilax glabra with reddish brown and off-white colored cross section].

There are two types of decoction of Smilax glabra due to its reddish brown or off-white colored cross section. These two kinds of decoction were found that they have large difference in anti-inflammatory effects and chemical constituents in the preliminary experiments. Comparing and analyzing the content of total tannin in these two kinds of decoction of S. glabra from 28 areas by UV-Vis spectrophotometry were first used to provide some experimental and theoretical development and utilization of this medicinal resource and quality control. Also, the sample recovery test required in Chinese Pharmacopoeia was improved by adding tannic acid instead of gallic acid to samples.

[Comparison of anti-inflammatory effect and analysis of astilbin red and white transverse section Smilax glabra in 28 collection sites].

OBJECTIVE: Astilbin in 28 Smilax glabra (red and white cross-section) from different sources was determined by HPLC. Pharmacodynamics and component of S. glabra was investigated through inflammation experiment (penetration type). METHOD: The analysis was performed on a Hypersil ODS2 column (4.6 mm x 250 mm, 5 microm) with the mobile phase of acetonitrile and 0. 1% acetic acid aqueous (21: 79) at a flow rate of 1.0 mL x min(-1). The detection wavelength was 291 nm, and the column temperature was 25 degrees C. Anti-inflammatory effect was compared from two type cross-section of Smilax glabra in capillary permeability experiment. RESULT: Linear correlation was good in the range of 0.003 379-4.004 microg, and the average recoveries were 100.1%, 101.9%, 99.3%, respectively. The content of astilbin in white and red transverse section were 0.19%-2.46% and 2.10%-5.92%, respectively. Anti-inflammatory efficiency of sectioned red and white were were 21% and 32%, respectively. CONCLUSION: Astilbin content and anti-inflammatory effect is significantly different between red and white transverse section of S. glabra, the content of astilbin is not positively related with anti-inflammatory effect.

UPLC-QTOF-MS identification of metabolites in rat biosamples after oral administration of Dioscorea saponins: a comparative study.

ETHNOPHARMACOLOGICAL RELEVANCE: Among the 49 species of the genus Dioscorea distributed in China, Dioscorea nipponica Makino (DN), Dioscorea panthaica Prain et Burkill (DP), and Dioscorea zingiberensis C. H. Wright (DZ) possess more or less similar traditional therapeutic actions, such as activating blood, relieving pain, and dispersing swelling; they have been used as folk medicine in China since 1950s. The modern pharmaceutical industry has developed these three species as herbal medicines that have been used for decades for treating cardiovascular diseases. However, there is no available information in the literature explaining how their chemical components are converted and interrelated in vivo to support their efficacies. The present study aimed to a) compare the metabolic profiles of saponins from DN, DP and DZ, which are considered to be their bioactive components, and b) to compare the changes in sustained levels of metabolites from rat biosamples. MATERIAL AND METHODS: Total saponins (TS) from each of the three species, and four individual saponins, namely protodioscin (PD), pseudoprotodioscin (PSD), dioscin (DC) and diosgenin (DG), were given to rats by oral administration. Chemical profiles of the rats' plasma, urine and feces were monitored 1-36 h. A UPLC-QTOF-MS based method was performed to identify the absorbed constituents and their metabolic products in rat biosamples (i.e., blood, urine, and feces); the ratio of peak area of major saponins to that of internal standard was calculated and plotted versus time to characterize the sustained levels of saponins in biosamples. RESULTS: Totally 10 saponin-related compounds were detected in rat plasma, 10 in rat urine and 18 in rat feces. The results indicated that formation of diosgenin by desugarization was the main pathway by which steroidal glycosides were metabolized. Other types of bio-transformation were found among glycosides and aglycones, such as ring cyclization through loss of 26-O-glucosyl, substitution of ß-D-glucopyranosyl for α-L-rhamnopyrannosyl, hydrogenation of diosgenin at 5(6)-double bond, and hydration of 20(22)-double bond. Generally, the metabolic profiles of DN and DP were shown to be quite similar, but different from that of DZ. However, some particular similarities and connections were found among these three TS. Diosgenin was one of the main metabolites commonly found in plasma and feces (excluding urine), from all groups receiving different TS, as well as individual saponins; this is likely to be one of the bioactive constituents playing an essential role in cardioprotective efficacy. Furostane-type saponins in TS of DN, DP or DZ, such as PD, protogracillin, parvifloside, protodeltonin and protobioside, showed fast absorption into blood (<1h), but were maintained for a relatively short period (mostly<8h), while the spirostane-type saponin and sapogenin (DC and DG, respectively), were absorbed into circulation more slowly (>1h), but increased gradually and lasted longer (>36h). These two patterns suggest that the therapeutic effect of these Dioscorea saponins is achieved through a complex, multi-step process over time. In addition, it appears that PD, PSD, and DC contained in DN and DP were transformed into certain glycosides originally found in DZ but not in DN or DP (protodeltonin, deltonin, trillin, and progenin II), which might indicate another linkage among these three species. CONCLUSION: These similarities and connections described above constitute evidence supporting similarity in efficacy of these three herbs from the perspective of metabolism. The UPLC-QTOF-MS based method is accurate and efficient for analyzing metabolic changes in rat biosamples over time.

Cardioprotective effect of total saponins from three medicinal species of Dioscorea against isoprenaline-induced myocardial ischemia.

ETHNOPHARMACOLOGICAL RELEVANCE: As folk medicines used in China since 1950s, Dioscorea nipponica Makino (DN), D. panthaica Prain et Burkill (DP), and D. zingiberensis C.H. Wright (DZ) are regarded as having more or less the same traditional therapeutic actions, such as activating blood, relieving pain, and dispersing swelling. It is noteworthy that, of the 49 species of the genus Dioscorea distributed in China, based on such traditional efficacies, only these three have been further developed as effective single-herb medicines for treating cardiovascular diseases by the modern pharmaceutical industry. In our previous study, it was found that the chemical compositions of DN and DP were similar, and both were distinct from that of DZ. Hence, whether their different chemical profiles support their anti-IHD (ischemic heart disease) activity in common still needs to be answered. So far it is still unknown whether the efficacies of these three herbs act via similar mechanism and whether they possess comparable therapeutic efficacy for experimental myocardial ischemia (MI). AIM OF THE STUDY: The present study aimed to further investigate the underlying mechanisms with respect to antioxidative stress activity by which these Dioscorea spp. attenuate MI, and to compare the therapeutic effect of total saponins from these three species on myocardial antioxidant levels and myocardial histology. MATERIAL AND METHODS: The serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), total superoxide dismutases (SOD), catalase (CAT), glutathione peroxidase (GPx), the total antioxidant capacity (T-AOC), and malondialdehyde (MDA), as well as myocardial histology, were compared among rat groups administered with total saponins (TS) of DN, DP or DZ (abbreviated as DNTS, DPTS and DZTS, respectively). The rats experienced myocardial ischemia induced by isoprenaline (ISO) injection; the test solutions (DNTS, DPTS, DZTS) were administered either after the ISO injection, or both before and after. RESULTS: Compared with the model group (ISO injection only), TS groups exhibited significantly reduced activities of CK, LDH and AST, lowered level of MDA, and increased activities of SOD, CAT, GPx and T-AOC; heart tissues from TS groups revealed less severe histological damage. The cardioprotective efficacy of these three Dioscorea TS for rat MI was closely comparable based on the above observations. CONCLUSION: The findings of the present study provide evidence that the anti-MI effect of DNTS, DPTS and DZTS can be attributed to the increase of myocardial antioxidant levels and decrease of lipid peroxidation formation, and the closely comparable results observed from these three Dioscorea saponins thereby explains the similarity in their clinical efficacy as anti-MI drugs.