Response surface methodology-optimized extraction of flavonoids with antioxidant and antimicrobial activities from the exocarp of three genera of coconut and characterization by HPLC-IT-TOF-MS/MS.

Response surface methodology optimization based on central composite design was applied to extract flavonoids from the exocarp of three coconut genera. The antioxidant and antimicrobial activities and structures of the flavonoids were determined. The results indicated that the optimal extraction conditions were ethanol concentration, 60%; temperature, 50 ℃; time, 90 min; liquid/material ratio, 40 mL/g; ultrasonic power, 150 W. Under these conditions, the yields of green, red and yellow coconut exocarp were 366.03 ± 7.57, 596.38 ± 10.32, and 403.78 ± 5.56 mg rutin/g powder, respectively. The flavonoids exhibited eminent DPPH and ABTS radical scavenging activities with IC50 values of 0.01-0.02 mg/mL. At a concentration of 2 mg/mL, they exhibited antimicrobial activity against Vibrio parahaemolyticus, Listeria monocytogenes, Escherichia coli, Staphylococcus aureus, Salmonella and Pseudomonas aeruginosa. In total, 17 flavonoids and 5 phenolic acids were characterized by UPLC-IT-TOF-MS/MS; among them, catechin, kaempferol, and quercetin were abundant. Yellow coconut had a distinct flavonoid spectrogram from other genera and contained more methoxy flavonoids.

Crataegus Special Extract WS 1442 Effects on eNOS and microRNA 155.

Increased expression of microRNA 155 (miR-155) results in a decrease in endothelial nitric oxide synthase (eNOS) expression and impaired endothelial function. Factors that have been shown to increase expression of miR-155 may be mitigated by WS 1442, an extract of hawthorn leaves and flowers (Crataegus special extract) that contains a range of pharmacologically active substances including oligomeric proanthocyanidins and flavonoids. The purpose of this study is to determine the effect of WS 1442 on the expression of miR-155 and eNOS in the presence of tumor necrosis factor (TNF-α). Human umbilical vein endothelial cells (HUVECs) were studied after the exposure to TNF-α, with or without simvastatin (positive control) and WS 1442. The expression levels of eNOS, phosphorylated eNOS, and miR-155 in the different HUVEC treatment groups were determined by western blot and quantitative real-time polymerase chain reaction, respectively. To evaluate the effect of WS 1442 on the eNOS activity, the medium and intracellular nitrate/nitrite (NO) concentrations were also analyzed using a colorimetric Griess assay kit. The results demonstrated that TNF-α upregulated miR-155 expression and decreased eNOS expression and NO concentrations. WS 1442 also increased miR-155 expression and decreased eNOS expression but, unlike TNF-α, increased phosphorylated eNOS expression and NO concentrations. Surprisingly, WS 1442 increased miR-155 expression; however, WS 1442 mitigated the overall negative effect of miR-155 on decreasing eNOS expression by increasing expression of phosphorylated eNOS and resulting in an increase in NO concentrations. In the setting where miR-155 may be expressed, WS 1442 may offer vascular protection by increasing the expression of phosphorylated eNOS.

Two new dammarane-type triterpenoid saponins from ginseng medicinal fungal substance.

Two new triterpenoid saponins, namely ginsenoside Re8 (1) and notoginsenoside ST-8 (2), were isolated from ginseng medicinal fungal substance. Their structures were elucidated by spectroscopic data and chemical analysis.

Two new dammarane-type triterpenoid saponins from notoginseng medicinal fungal substance.

Two new dammarane-type triterpenoid saponins, namely ginsenoside Rk6 (1) and ginsenoside-Rh22 (2), were isolated from notoginseng medicinal fungal substance. The structures of 1 and 2 were established as 3ß,6α,12ß,26-tetrahydroxydammar-20(21),24(25)(E)-diene-6-O-ß-D-glucopyranoside and 3ß,6α, 20(S)-trihydroxy-12(R),23(R)-expoxy-13(S),17(S)-dammar-24-ene-6-O-ß-D-glucopyranoside on the basis of spectroscopic analysis and chemical analysis, respectively.

Two new triterpenoid saponins from ginseng medicinal fungal substance.

Two new dammarane-type triterpenoid saponins, namely ginsenosides Rb4 (1) and Rb5 (2), were isolated from ginseng medicinal fungal substance. The structures of 1 and 2 were established as 3ß,12ß,20(S)-trihydroxydammar-24(25)-ene-3-O-[α-d-glucopyranosyl-(1→4)-ß-d-glucopyranosyl-(1→2)-ß-d-glucopyranosyl]-20-O-ß-d-glucopyranoside and 3ß,12ß,20(S)-trihydroxydammar-24(25)-ene-3-O-[α-d-glucopyranosyl-(1→4)-α-d-glucopyranosyl-(1→4)-ß-d-glucopyranosyl-(1→2)-ß-d-glucopyranosyl]-20-O-ß-d-glucopyranoside on the basis of spectroscopic analysis and chemical analysis, respectively.

Metabolomics and its application to the evaluation of the efficacy and toxicity of traditional Chinese herb medicines.

Traditional Chinese herb medicines (TCHMs) have been used in the treatment of a variety of diseases for thousands of years in Asian countries. The active components of TCHMs usually exert combined synergistic therapeutic effects on multiple targets, but with less potential therapeutic effect based on routine indices than Western drugs. These complex effects make the assessment of the efficacy of TCHMs and the clarification of their underlying mechanisms very challenging, and therefore hinder their wider application and acceptance. Metabolomics is a crucial part of systems biology. It allows the quantitative measurement of large numbers of the low-molecular endogenous metabolites involved in metabolic pathways, and thus reflects the fundamental metabolism status of the body. Recently, dozens of metabolomic studies have been devoted to prove the efficacy/safety, explore the underlying mechanisms, and identify the potential biomarkers to access the action targets of TCHMs, with fruitful results. This article presents an overview of these studies, focusing on the progress made in exploring the pharmacology and toxicology of various herbal medicines.

Development of a systematic approach to rapid classification and identification of notoginsenosides and metabolites in rat feces based on liquid chromatography coupled triple time-of-flight mass spectrometry.

The present work contributes to the development of a powerful technical platform to rapidly identify and classify complicated components and metabolites for traditional Chinese medicines. In this process, notoginsenosides, the main active ingredients in Panaxnotoginseng, were chosen as model compounds. Firstly, the fragmental patterns, diagnostic product ions and neutral loss of each subfamily of notoginsenosides were summarized by collision-induced dissociation analysis of representative authentic standards. Next, in order to maximally cover low-concentration components which could otherwise be omitted from previous diagnostic fragment-ion method using only single product ion of notoginsenosides, a multiple product ions filtering strategy was proposed and utilized to identify and classify both non-target and target notoginsenosides of P.notoginseng extract (in vitro). With this strategy, 13 protopanaxadiol-type notoginsenosides and 30 protopanaxatriol-type notoginsenosides were efficiently extracted. Then, a neutral loss filtering technique was employed to trace prototype components and metabolites in rats (in vivo) since diagnostic product ions might shift therefore become unpredictable when metabolic reactions occurred on the mother skeleton of notoginsenosides. After comparing the constitute profiles in vitro with in vivo, 62 drug-related components were identified from rat feces, and these components were classified into 27 prototype compounds and 35 metabolites. Lastly, all the metabolites were successfully correlated to their parent compounds based on chemicalome-metabolome matching approach which was previously built by our group. This study provided a generally applicable approach to global metabolite identification for the complicated components in complex matrices.

An ex vivo approach to botanical-drug interactions: a proof of concept study.

ETHNOPHARMACOLOGICAL RELEVANCE: Botanical medicines are frequently used in combination with therapeutic drugs, imposing a risk for harmful botanical-drug interactions (BDIs). Among the existing BDI evaluation methods, clinical studies are the most desirable, but due to their expense and protracted time-line for completion, conventional in vitro methodologies remain the most frequently used BDI assessment tools. However, many predictions generated from in vitro studies are inconsistent with clinical findings. Accordingly, the present study aimed to develop a novel ex vivo approach for BDI assessment and expand the safety evaluation methodology in applied ethnopharmacological research. MATERIALS AND METHODS: This approach differs from conventional in vitro methods in that rather than botanical extracts or individual phytochemicals being prepared in artificial buffers, human plasma/serum collected from a limited number of subjects administered botanical supplements was utilized to assess BDIs. To validate the methodology, human plasma/serum samples collected from healthy subjects administered either milk thistle or goldenseal extracts were utilized in incubation studies to determine their potential inhibitory effects on CYP2C9 and CYP3A4/5, respectively. Silybin A and B, two principal milk thistle phytochemicals, and hydrastine and berberine, the purported active constituents in goldenseal, were evaluated in both phosphate buffer and human plasma based in vitro incubation systems. RESULTS: Ex vivo study results were consistent with formal clinical study findings for the effect of milk thistle on the disposition of tolbutamide, a CYP2C9 substrate, and for goldenseal׳s influence on the pharmacokinetics of midazolam, a widely accepted CYP3A4/5 substrate. Compared to conventional in vitro BDI methodologies of assessment, the introduction of human plasma into the in vitro study model changed the observed inhibitory effect of silybin A, silybin B and hydrastine and berberine on CYP2C9 and CYP3A4/5, respectively, results which more closely mirrored those generated in clinical study. CONCLUSIONS: Data from conventional buffer-based in vitro studies were less predictive than the ex vivo assessments. Thus, this novel ex vivo approach may be more effective at predicting clinically relevant BDIs than conventional in vitro methods.

[Mating system and causes of low fructification rate by self-pollination of Codonopsis pilosula].

OBJECTIVE: To study four kinds of germplasm resources of Codonopsis pilosula and provide the basal mating systems data for the breeding and cultivation of C. pilosula. METHOD: 0.5% TTC (2,3,5-triphenyl tetrazolium chloride) solution was used for the pollen viability test and benzidineand-H2O2 [1% benzidine in 60% ethanol,hydrogen peroxide (3%), and water, 4:11:22] was used for estimation of the stigma receptivity. The mating systems were tested by out crossing index (OC1), pollen-ovule ratio (P/O) and pollination by bagged and emasculated in the field. RESULT: The pollen viability of C. pilosula reached highly about 80% when the pollen staying in the anther, 2-3 days before the petals opening. The anther began scattering pollen before the day of the petals opening, the pollen viability was the highest about 95%, the pollen stick thickly aroud the stigma and quickliy lost in the next day. The stigma life-span was about 4-5 days, the optimal time for pollination was the first day of the petals opening, when the stigma was highly sticky and yellow. The value of out crossing index (OC1) was 4, pollen-ovule ratio was between 104.84-185.75. The natural fructification rate of cross-pollination by emasculated-treatment was 25.6% 42.4%. The fructification rate and compatible index of self-pollination by bagged- treatment were about 3.3%-6.7% and 3.0-21.8. CONCLUSION: The mating system of C. pilosula is mixed with self-pollination and cross-pollination, prone to cross-pollination. The compatibility of self-pollination is high. The difference of maturing period of pistil and stamen and the lack of polen amount cause low fructification rate of self-pollination.

The soybean isoflavonoid equol blocks ritonavir-induced endothelial dysfunction in porcine pulmonary arteries and human pulmonary artery endothelial cells.

HIV protease inhibitor (PI) ritonavir (RTV) may cause vascular injury through oxidative stress. Our purpose in this study was to determine whether equol, a soy isoflavone, could prevent RTV-induced endothelial dysfunction in porcine pulmonary arteries and in human pulmonary artery endothelial cells (HPAEC). Fresh porcine pulmonary artery rings were treated with 15 micromol/L of RTV and/or equol in concentrations of 0.1, 1, and 10 micromol/L for 24 h. A control was set with no amount of equol or RTV administered. Based on myograph tension analysis, RTV significantly reduced endothelium-dependent relaxation in response to bradykinin in the artery rings compared with untreated vessels, whereas the antioxidant equol effectively reversed the RTV effect in a concentration-dependent manner. RTV also reduced the contraction of artery rings in response to thromboxane A(2) analogue U46619 and this reduction was blocked by equol. In addition, RTV treatment significantly reduced endothelial nitric oxide synthase (eNOS) expression in both porcine pulmonary arteries and HPAEC, whereas equol effectively blocked RTV-induced eNOS downregulation. Furthermore, RTV significantly increased superoxide anion production, whereas equol reversed this effect of RTV in porcine pulmonary arteries. Thus, the antioxidant equol effectively protects vascular function from the detrimental effects of HIV PI RTV in both porcine pulmonary arteries and HPAEC via a reduction in the vasomotor dysfunction, eNOS downregulation, and oxidative stress induced by RTV. These novel data suggest that equol may have a clinical application in preventing HIV-associated cardiovascular complications.

Ginsenoside Rb1 attenuates homocysteine-augmented guidewire injury-induced intimal hyperplasia in mice.

BACKGROUND: Intimal hyperplasia (IH) is the primary cause for post-angioplasty restenosis. The purpose of this study is to investigate the effects of homocysteine (Hcy) and ginsenoside Rb1 (Rb1) on IH using a guidewire injury animal model. METHODS: In 12-wk-old C57BL/6J mice, the left common carotid artery (CCA) was denudated with a guidewire and the right CCA was used as the uninjured control. They were treated with saline (NS), Hcy, Rb1, or Hcy + Rb1 for 4 wk prior to sacrifice. Animals were sacrificed at 4, 6, or 8 wk. Both CCAs were harvested and intimal-medium thickness (IMT) ratios were calculated. Local macrophage distribution was also studied. RESULTS: Histology analyses demonstrated consistent internal elastic lamina disruption and focal IH in the injured CCA segments. The degree of IH correlated to the lengths of time following injury. Hcy treated group had significant increase in IMT compared with the NS group (P < 0.05), while Rb1 group was similar to the NS group. In addition, Hcy + Rb1 group showed significant improvement in IMT compared with Hcy group (P < 0.01). Furthermore, Hcy significantly increased local macrophage content as compared with either lesion alone or Rb1 treated animals. CONCLUSIONS: Our study showed that Hcy increased the degree of IH and macrophage content in the injured CCA and that Rb1 attenuated these adverse effects. These changes might be mediated through antioxidative effects of Rb1. Our data suggests a potential clinical application of ginseng in controlling Hcy-related vascular injuries.