Macrophage ß-arrestin-1 deteriorates DSS-induced colitis through interaction with NF-κB signaling.

ß-arrestin-1 has been demonstrated to participate in the regulation of inflammatory reactions in several diseases. Thus, this study aimed to investigate the role of macrophage ß-arrestin-1 in the pathogenesis and progression of ulcerative colitis (UC). A myeloid ß-arrestin-1 conditional knockout mouse model was generated to explore the role of macrophage ß-arrestin-1. DSS was employed for the establishment of an ulcerative colitis mouse model, using TNF-α as an inflammatory stressor in vitro. The expression level of ß-arrestin-1 was detected via western blot and immunofluorescence assays, whilst disease severity was evaluated by clinical score and H&E staining in the DSS-induced colitis model. In the in vitro experiments, the levels of inflammatory cytokines were examined using real-time PCR. NF-κB activation was detected through the double luciferase reporter system, western blot, and electrophoretic mobility shift assay (EMSA). BAY11-7082 was used to inhibit NF-κB activation. Our results exposed that the level of ß-arrestin-1 was increased in monocytes/macrophages derived from DSS-induced colitis mice or under the TNF-α challenge. Moreover, conditionally knocking out the expression of myeloid ß-arrestin-1 alleviated disease severity, while knocking out the expression of ß-arrestin-1 decreased the levels of inflammatory cytokines. Additionally, NF-κB was identified as a central regulatory element of ß-arrestin-1 promoter, and using BAY11-7082 to inhibit NF-κB activation lowered the level of ß-arrestin-1 under TNF-α challenge. ß-arrestin-1 led to the activation of the NF-κB signaling pathway by enhancing binding to IκBα and IKK under the TNF-α challenge. Taken together, our findings demonstrated macrophage ß-arrestin-1 contributes to the deterioration of DSS-induced colitis through the interaction with NF-κB signaling, thus highlighting a novel target for the treatment of UC.

Depletion of ß-arrestin-1 in macrophages enhances atherosclerosis in ApoE-/- mice.

Autophagy in atherosclerotic plaque macrophage contributes to the alleviation of atherosclerosis through the promotion of lipid metabolism. ß-arrestins are multifunctional proteins participating various kinds of cellular signaling pathways. Here we aimed to determine the role of ß-arrestin-1, an important member of ß-arrestin family, in atherosclerosis, and whether autophagy was involved in this process. ApoE-/-ß-arrestin-1fl/flLysM-Cre mice were created through bone marrow transplantation for the atherosclerosis model with conditional myeloid knocking out ß-arrestin-1. Bone marrow-derived macrophages (BMDMs) were used for the in vitro studies. Oil red O staining was used to detect the lesional area. F4/80, Masson trichrome and picro-Sirius red staining were applied for the determination of plaque stability. Real-time PCR was used for the detection of levels of lipid metabolism-related receptors. Electron microscopy and tandem fluorescent mRFP-GFP-LC3 plasmid was applied to test autophagy level. We found that ß-arrestin-1 was highly increased in expression in plaque macrophage on the occurrence of atherosclerosis. Conditional myeloid knocking out ß-arrestin-1 largely promotes plaque formation and vulnerability. In murine macrophage with lipid loading, knocking down ß-arrestin-1 enhanced foam cell formation and levels of plasma and cellular cholesterol, while overexpressing ß-arrestin-1 led to the opposite effects. The alleviative effects induced by macrophage ß-arrestin-1 in atherosclerosis were involved in autophagy, based on the reduction of autophagy level with the knocking down of macrophage ß-arrestin-1 and administration of autophagy inhibitors which largely attenuated the decreasing effect on foam cell formation. Our results demonstrated for the first time that macrophage ß-arrestin-1 protected against atherosclerosis through the induction of autophagy.

[Spatial Distribution, Source Analysis, and Health Risk Assessment of Metal Elements in Karst Water in Southeastern Chongqing].

Dispersed karst water is an important water supply source, or even the only water supply source, for some districts and counties in Chongqing City. It is particularly necessary to understand the distribution characteristics of metal elements in karst water and the health risks exposed. In this study, the scattered karst water in the southeastern part of Chongqing was taken as the main research object, and the concentrations of Al, Cu, Pb, Zn, Cr, Cd, Ni, Mn, As, and Hg in 42 groups of karst spring water samples were determined. The spatial distribution of metal elements with a high detection rate was revealed using the ordinary kriging interpolation method, and the spatial distribution characteristics, sources, and health risks of metal elements were analyzed using multivariate statistical methods and health risk models. The results showed that the quality of dispersed karst water in southeastern Chongqing was generally good, and the spatial scale variability in the occurrence of metal elements in karst water was strong, especially for Ni and As. The sources of Cu, Pb, As, Zn, and Cr were mainly affected by the regional geological background; Al and Mn were mainly affected by human industrial, agricultural, and mining activities; and Ni was affected by both the natural background and human activities. The total health risk of exposure through the drinking route was higher than that of the skin infiltration route, which was the main exposure route of the human body. The total health risk of children exposed through the drinking route was higher than that of adults, and the total health risk of adults exposed through the skin infiltration route was higher than that of children. It is worth noting that Cr was the determinant of total health risk. From the perspective of drinking water safety, local residents need to pay certain attention to water quality when drinking distributed karst groundwater, in order to reduce the health risk of the population.

[Pollution and Irrigation Applicability of Surface Water from Wet, Normal, and Dry Periods in the Huixian Karst Wetland, China].

To reveal the hydrochemical characteristics of karst wetland located in a subtropical area and at lower elevations in China, 27 surface water samples were collected during three periods (wet, normal, and dry) in the Huixian karst wetland to investigate the distributions, pollution, and irrigation application of 12 inorganic ions and 10 heavy metals. Based on their concentrations, the Nemerow index and the four evaluation systems of the sodium adsorption ratio (SAR), sodium concentration (SC), permeability index (PI), and residual sodium carbonate (RSC) were applied to evaluate the pollution characteristics and irrigation application. It was found that the water type in this area was Ca2+-HCO3- and weakly alkaline. Regarding the 12 inorganic ions and 10 heavy metals, NH4+ exceeded the Chinese standards for drinking water with an exceedance rate of 25.93%, and the exceedance rates of Al, Mn, and Hg were 11.11%, 44.44%, and 37.04%, respectively. The spatiotemporal scaling effect on inorganic ions was lower than that of heavy metals, and the distributions of the inorganic ions and heavy metals were in the order of wet period > normal period > dry period. However, the surface water quality in the Huixian karst wetland was generally well-protected based on the pollution assessment. The Nemerow index ranged from 0.75 to 2.69, which recognized the main pollution contributors as NH4+, Mn, Al, and Hg with the contamination grade from slight pollution to moderate pollution, especially in the core area during the wet period. According to the limits of standards for irrigation water quality and environmental quality for surface water, as well as the evaluation results of the SAR, SC, PI, and RSC, the surface water in the Huixian karst wetland was generally suitable for irrigation, and the water quality in the dry period was better than that in the wet and normal periods. The surface water from site PH1 during the normal period with 19.1 µg·L-1 of Hg and site FH8 during the wet period with 13.7 mg·L-1 of NH4+ were not suitable for agricultural irrigation.

[Major Ionic Characteristics and Factors of Karst Groundwater at Huixian Karst Wetland, China].

To investigate the major ionic chemical characteristics and seasonal variations, 27 groundwater samples were collected from the wet season, flat season, and dry season during 2018-2019 in the Huixian Karst wetland, which is the largest low-altitude karst wetland in China. The single pollution standard index was applied to evaluate the groundwater pollution during different periods, and the major ionic factors of the karst groundwater were analyzed using the statistical analysis method, Gibbs diagram, and ion ratio. The results revealed that the groundwater samples were a weakly alkaline fresh water that were rich in Ca2+ and HCO3-. The average concentrations of the primary ions followed the order of flat season > wet season > dry season; meanwhile, the water quality in the dry season was better than that in the wet and flat seasons. The K+ and NO3- in the karst groundwater were mostly affected by the spatial distributions of the aquifers, and the Mg2+, SO42-, NO2-, NH4+, and TDS were related to the space-season scale. Na+, Ca2+, HCO3-, and Cl- were relatively stable ions in the karst groundwater. The hydrochemical characteristics were primarily determined by carbonate rock dissolution and were found to be the HCO3-Ca type, which accounted for 77.78%, 77.78%, and 88.89% in the wet season, flat season, and dry season, respectively. The karst groundwater was predominantly polluted by SO42-, NO3-, and NO2-; particularly, NO3- exhibited serious pollution points, and SO42- had heavy pollution points in the wet and flat seasons. The chemical composition of the karst groundwater was controlled mostly by water-rock interactions. Ca2+ and HCO3- primarily came from calcite dissolution, and the high concentrations of Mg2+ and SO42- in a few number of points were controlled by dolomite, dolomitic limestone, and pyrite. K+, Na+, SO42-, NO3-, and Cl- partly came from atmospheric precipitation, and Na+ and Cl- partly came from human activities; K+ was related to potash fertilizer, and the main source of NO3- was chemical fertilizer.

[Metal Pollutions and Human Health Risks in Groundwater from Wet, Normal, and Dry Periods in the Huixian Karst Wetland, China].

The concentrations of 10 metals (Cd, Cr, As, Al, Cu, Pb, Zn, Mn, Hg, and Fe) in 27 groundwater samples collected during different periods (wet, normal, and dry) in the Huixian Karst wetland, the largest subtropical low-altitude karst wetland in China, were detected and analyzed to investigate pollution and health risks. The pollution characteristics, distribution, and health risks of the metals in groundwater were revealed by a comprehensive pollution assessment, multivariate statistical analysis, and health risk assessment model, respectively. The results showed that the average concentrations of metals in groundwater were followed the order of Mn > Fe > Zn > Al > Hg > Cr > Cu > Cd > As > Pb. The maximum concentration of Mn (1022.00 µg·L-1) was found in the wet season, while that of Hg (42.40 µg·L-1) was found in the normal season, and both were over the corresponding standard limits. The results of the pollution assessment indicated that only Mn pollution reached level Ⅵ in the wet season, while Cd, Al, Zn, and Fe pollution were at the level of Ⅲ. Only the Hg pollution level reached level Ⅵ while Al pollution reached the level of Ⅲ in the normal water period. According to the above results, the water quality in the dry season was better than that in the wet and normal seasons in terms of the 10 metals. The concentrations of Zn, Cd, Mn, and Al in groundwater were affected by human activities, while the time-scale characteristics of these were not obvious. The concentrations of As, Fe, Cu, and Cr were all affected by human activities and the time-scale, while the concentrations of Hg and Pb were mainly manifested in time-scale characteristics. The results of the health risk assessment of the water due to drinking and the skin penetration pathway indicated that the total health risks followed the order of normal season > the wet season > the dry season. The carcinogenic risks caused by Cr for adults and children through drinking pathway in the wet season (8.03×10-5 a-1 and 8.76×10-5 a-1), normal season (1.15×10-4 a-1 and 1.26×10-4 a-1),and dry season (8.72×10-5 a-1 and 9.51×10-5 a-1) exceeded the maximum allowed level (5.0×10-5 a-1) in all periods. Hence, Cr was the main metal element that caused carcinogenic risks. For the sake of drinking water safety, the concentrations of Mn, Hg, and Cr in groundwater should be controlled before drinking.

Cause and control of Radix Ophiopogonis browning during storage.

In the storage of Radix Ophiopogonis, browning often happens to cause potential risk with regard to safety. Previously few reports investigate the browning of Radix Ophiopogonis. In this research, the causes and mechanisms of the browning of Radix Ophiopogonis were preliminarily elucidated. Content determination by high-performance liquid chromatography (HPLC) and spectrophotometry, enzyme activity determination by colorimetry, and morphological observation by electron microscopy were performed in the present study. Uniform design and three-dimensional response surfaces were applied to investigate the relationship between browning and storage factors. The cortex cell wall of browned Radix Ophiopogonis was ruptured. Compared with the normal Radix Ophiopogonis, cellulase and polyphenol oxidase enzymes were activated, the levels of 5-hydroxymethylfurfural (5-HMF), total sugars, and reducing sugars were increased, while the levels of polysaccharides and methylophiopogonanone A were decreased in browned Radix Ophiopogonis. The relationship between the storage factors and degree of browning (Y) could be described by following correlation equation: Y = - 0.625 4 + 0.020 84 × X3 + 0.001 514 × X1 × X2 - 0.000 964 4 × X2 × X3. Accompanied with browning under storage conditions, the chemical composition of Radix Ophiopogonis was altered. Following the activation of cellulase, the rupture of the cortex cell wall and the outflow of cell substances flowed out, which caused the Radix Ophiopogonis tissue to become soft and sticky. The main causes of the browning were the production of 5-HMF, the activation of polyphenol oxidase, Maillard reactions and enzymatic browning. Browning could be effectively prevented when the air relative humidity (HR), temperature, and moisture content were under 25% RH, 12 °C and 18%, respectively.

Steroidal sapogenins and glycosides from the fibrous roots of Polygonatum odoratum with inhibitory effect on tissue factor (TF) procoagulant activity.

Six new spirostane glycosides (1-6), named polygodosides A-F, one new furostanol glycoside, polygodoside G (7), one new cholestane glycoside, polygodoside H (8), and one new steroidal sapogenin, polygodosin A (9), together with thirteen known compounds (10-22) were isolated from a 90% MeOH extract of the fibrous roots of Polygonatum odoratum (Mill.) Druce. The structures of new compounds were elucidated by extensive 1D and 2D NMR spectroscopic analyses and mass spectrometry. The effects on TF procoagulant activity in THP-1 cells were tested for most of the compounds.

Simultaneous determination of puerarin, daidzin, daidzein, paeoniflorin, albiflorin, liquiritin and liquiritigenin in rat plasma and its application to a pharmacokinetic study of Ge-Gen Decoction by a liquid chromatography-electrospray ionization-tandem mass spectrometry.

A liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method was developed and validated for simultaneous determination of seven constituents including puerarin, daidzin, daidzein, paeoniflorin, albiflorin, liquiritin and liquiritigenin in rat plasma using schisandrin as the internal standard (IS). The plasma samples were pretreated by a one-step direct protein precipitation with acetonitrile. The chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of acetonitrile and water (containing 0.1% formic acid and 5mM ammonium acetate). All analytes and IS were quantitated through electrospray ionization in positive ion multiple reaction monitoring (MRM) mode. The mass transitions were as follows: m/z 417.5→297.2 for puerarin, m/z 417.1→255.2 for daidzin, m/z 255.2→152.4 for daidzein, m/z 498.1→179.3 for paeoniflorin, m/z 481.1→197.3 for albiflorin, m/z 436.2→257.3 for liquiritin, m/z 257.2→137.3 for liquiritigenin and m/z 415.0→384.2 for IS, respectively. All calibration curves exhibited good linearity (r>0.9979) over a wide concentration range for all components. The intra-day and inter-day precisions (RSD) at three different levels were both less than 14.3% and the accuracies (RE) ranged from -13.2% to 14.8%. The extraction recoveries of the seven compounds ranged from 72.9% to 117.4%. The validated method was successfully applied to pharmacokinetic study of the seven components in female rat plasma after oral administration of Ge-Gen Decoction aqueous extract.

Identification and determination of the major triterpenes in Rhizoma Alismatis by HPLC-evaporative light scattering detection and HPLC/electrospray ionization-MS.

A simple and reliable HPLC-evaporative light scattering detection method was established for the determination of seven triterpenes in Rhizoma Alismatis, a commonly used herbal medicine. HPLC coupled with electrospray ionization-MS was applied to identify the triterpenes. The positive ion mode was used in MS detection, and the fragmentation patterns of the analytes were proposed. The quantitative method was validated for linearity, sensitivity, precision, and accuracy. All calibration curves showed good linearity (r2 > 0.9943) within the test ranges. This method showed good reproducibility with intraday and interday variations of less than 3.39 and 5.20%, respectively. The mean recoveries ranged from 96.06 to 103.5%. The proposed method was successfully applied for the quantitative analysis of the triterpenes in samples from four different habitats. The results indicated great variation of the contents of these components among the samples, and the developed assay could be considered as a suitable quality evaluation method for Rhizoma Alismatis.

Screening and identifying the myocardial-injury protective ingredients from Sheng-Mai-San.

CONTEXT: Sheng-Mai-San (SMS) has been used for the treatment of cardiovascular disease for many years in China. OBJECTIVES: This study investigated the protective effects and active ingredients of SMS on myocardial injury (MI) in mice. MATERIALS AND METHODS: SMS and n-butanol extraction of SMS (SMS-Bu) were prepared and administered to ISO-treated mice once a day for 7 consecutive days. The doses were equivalent to the raw medicinal herbs of SMS 5.72, 2.86 and 1.43 g/kg/d, respectively. Propranolol was used as positive control. Serum biomarkers, histopathological and electrocardiographic were evaluated. RESULTS: Serum lactate dehydrogenase, creatine kinase and myeloperoxidase increased to 4473.6 ± 322.5, 950.0 ± 35.0 and 90.4 ± 12.2 U/L in the model group. SMS and SMS-Bu groups showed a decrease from 10 to 29% for lactate dehydrogenase and from 17 to 42% for creatine kinase, respectively. Both SMS and SMS-Bu significantly attenuated the myeloperoxidase activities (from 42 to 56%) and malondialdehyde levels (from 25 to 45%) compared with the model group. Decreased superoxide dismutase activities in ISO-treated mice were elevated from 19 to 59% when treated with SMS and SMS-Bu. These biochemical results were supported by electrocardiogram (ECG) and histopathological observations. Furthermore, 8 ginsenosides and 16 lignans were identified in SMS-Bu. CONCLUSION: These findings suggested that SMS-Bu was the mainly active fraction of SMS which exerted its beneficial effects on MI mainly through protecting myocardial tissue and reducing oxidative damage, and the ginsenosides and lignans may serve as active ingredients of SMS for the treatment of MI.

Protective effects and active ingredients of yi-qi-fu-mai sterile powder against myocardial oxidative damage in mice.

This study aims to evaluate the protective effects of Yi-Qi-Fu-Mai sterile powder (YQFM) on myocardial oxidative damage and tries to identify the active components responsible for its pharmacological benefits. YQFM and the n-butanol extract of YQFM (YQFM-Bu) were administered to ISO-induced myocardial injury mice. Left ventricle weight index and histopathological analyses were conducted. Serum enzymatic activities of lactate dehydrogenase (LDH), creatine kinase (CK) and superoxide dismutase (SOD), myeloperoxidase (MPO), and the levels of malondialdehyde (MDA) were also measured. Our results demonstrated that both YQFM and YQFM-Bu significantly restored the abnormal activities of CK, LDH, MPO, SOD, and the levels of MDA in ISO-induced myocardial injury mice, and these biochemical results were further supported by histopathological data. Our in vitro findings also confirmed that both YQFM and YQFM-Bu exhibit significant radical scavenging activity. Furthermore, the major active fractions of YQFM were identified by UPLC-MS/MS. Twenty-five ginsenosides and three lignans were identified from YQFM-Bu. These findings suggested YQFM-Bu is the major active fraction of YQFM with the ginsenosides and lignans as potential active components responsible for its protective effect against myocardial injury, and YQFM exerted its beneficial effects on myocardial injury mainly through inhibiting oxidative damage and maintaining the functional integrity of myocardial tissue.

HPLC-DAD-Q-TOF-MS/MS analysis and HPLC quantitation of chemical constituents in traditional Chinese medicinal formula Ge-Gen Decoction.

Ge-Gen Decoction (GGD) is a classical formula of traditional Chinese medicine. It is generally used for treating common cold, fever and influenza in China and South East Asia. In this study, a systematic method was established for the qualitative and quantitative analysis of the major constituents in GGD. For qualitative analysis, a method of liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS) was developed for identification of multi-constituents. Based on the UV spectra, retention time and MS spectra, sixty compounds in GGD extract were identified or tentatively characterized by comparing with reference substances or literatures. According to the qualitative results, a new quantitative analysis method of GGD was established by HPLC-DAD. Fourteen representative compounds unequivocally identified were chosen as marker components which were derived from five herbs in GGD excluding Zingiberis Rhizoma Recens and Jujubae Fructus. The analytical method was validated through intra- and inter-day precision, repeatability and stability, and the R.S.D. was less than 3.18%, 4.48%, 3.36% and 3.54%, respectively. The LODs and the LOQs for the analytes were less than 1.06 and 3.12µgmL(-1), respectively. The overall recoveries ranged from 94.8% to 105.6%, with the R.S.D. ranging from 0.68% to 3.23%. Then the new method was applied to determine twelve batches of GGD commercial products of three dosage forms. The results indicated that the new approach was applicable in the routine analysis and quality control of GGD products. The study might provide a basis for quality control of GGD, and further study of GGD in vivo.

Determination and pharmacokinetics of DT-13 in rat plasma by LC-MS.

A sensitive and specific LC-MS assay for DT-13 in rat plasma was developed. DT-13 is an active steroidal saponin present in Liriopes Radix and is developed as an anti-tumor drug candidate. The samples were extracted by acetonitrile-mediated plasma protein precipitation. The chromatographic separation was carried out using a Ultimate C(18) column (250 mm × 4.6 mm, i.d., 5 µm) with a mobile phase composed of acetonitrile: 5 mmol/L aqueous ammonium acetate (60:40, v:v). The method was validated and the specificity, linearity (r(2)=0.9980 within 10-1000 ng/mL), lower limit of quantitation (LLOQ, 10 ng/mL), precision (intra- and inter-day <12.3%), accuracy (93.4-106.3%), recovery (91.0 ± 4.7%) and stability were determined. The method was applied to the pharmacokinetic study of DT-13 in rat plasma after intravenous and intragastric administration. The results showed DT-13 underwent a prolonged absorption and slow elimination with a low oral bioavailability (5.51%) in rats.

Identification of multiple constituents in the traditional Chinese medicine formula Sheng-Mai San and rat plasma after oral administration by HPLC-DAD-MS/MS.

Sheng-Mai San (SMS), a traditional Chinese medicine formula, has been used for the treatment of cardiovascular disease in Asia over long period of time. While its effectiveness has been confirmed by clinical use, its active chemical constituents remain unclear. In this paper, an HPLC-DAD-MS/MS method is described for the efficient and rapid identification of the chemical constituents in SMS extract. MS/MS fragmentation behavior of authentic compounds was proposed for aiding the structural identification of the components. A total of 53 compounds were identified or tentatively characterized by comparing their retention times, UV and MS spectra with those of authentic compounds or literature data. HPLC/UV and MS techniques were employed to screen for the potential bioactive components in rat plasma after oral administration of SMS. Twenty-five compounds including 14 prototype components and 11 metabolites were detected in dosed rat plasma compared with blank rat plasma. This identification and structural elucidation of the chemical constituents in the medicine formula and rat plasma may provide important experimental data for further pharmacological and clinical research.

Effects of storage condition factors on fungal invasion of Radix Ophiopogonis.

Fungal invasion is a main factor leading to the loss of postharvest herbs during storage. To prevent fungal invasion and clarify the favorable conditions for the stability of herbs during the storage period, uniform design and three-dimensional response surfaces were applied to investigate the relationship of the mildew degree of Radix Ophiopogonis induced by prevalent fungal isolates and the storage factors including air relative humidity (X(1)), temperature (X(2)), and moisture content (X(3)), in laboratory studies. Mildew degree was evaluated by ergosterol assay of mold isolates through a high-performance liquid chromatography-diode array detector (HPLC-DAD) technique. As a result, storage conditions, which satisfied the following correlation equations established by the relationships between the storage factors and the biomass of mildew (Y), could effectively prevent fungal spoilage: Y = -0.2484084028 + 0.00030711966572 x X(3) x X(3) + 0.00018881361186 x X(1) x X(2) - 0.00029473040679 x X(2) x X(3)

Study on the radical scavengers in the traditional Chinese medicine formula shengmai san by HPLC-DAD coupled with chemiluminescence (CL) and ESI-MS/MS.

Shengmai San (SMS) has been used for the treatment of cardiovascular disease in Asia for long-time. It has been reported that SMS can protect against oxidative stress and tune immune response in patients. In this paper, a high performance liquid chromatography-diode array detection-chemiluminescence (HPLC-DAD-CL) method for on-line detection was successfully applied to screen antioxidants in complex SMS extracts. The H(2)O(2) scavenging activity of every fraction of SMS extracts was measured to evaluate each contribution to the total anti-oxidative activities of the prescription, followed by structures identification of the main active components by LC/MS/MS. HPLC-DAD-CL method was also applied for comparing the scavenging rates of main active components between the individual herbs and SMS. Interestingly, we found that the scavenging activities of main components detected in the individual herbs were different from those in SMS. The experimental data indicated that drug interactions during decocting process could result in the changes of the antioxidants' solubility. This study demonstrated that HPLC-DAD-CL on-line assay was a useful technique for rapidly screening and identifying bioactive components from complex multi-herbal prescription.

Homoisoflavonoids from the fibrous roots of Polygonatum odoratum with glucose uptake-stimulatory activity in 3T3-L1 adipocytes.

The EtOAc-soluble fraction of a 90% MeOH extract of the fibrous roots of Polygonatum odoratum was found to potentiate insulin-stimulated glucose uptake in differentiated 3T3-L1 adipocytes. Bioassay-guided fractionation yielded nine homoisoflavonoids (1-9), four of which were new (1-4), together with an isoflavone glycoside (10) and a flavanone glycoside (11). The structures of new compounds were elucidated on the basis of extensive 1D and 2D NMR spectroscopy, and the absolute configurations were deduced by CD spectra. All 11 compounds showed effects of sensitizing adipocytes for insulin in a cell-based glucose uptake assay using 3T3-L1 adipocytes. The results indicate that homoisoflavonoids may be potential insulin sensitizers.

Quality control of flavonoids in Ginkgo biloba leaves by high-performance liquid chromatography with diode array detection and on-line radical scavenging activity detection.

Flavonoids in plants used for the treatment of various cardiovascular, cancer diseases have been reported to possess potential protective effects against oxidative injury. Ginkgo biloba leaves, known for their antioxidant activity, were chosen for this study. In this paper, 12 flavonoids in G. biloba leaves were identified by HPLC-diode array detection (DAD)-electrospray ionization MS. HPLC-DAD coupled with chemiluminescence detection was used to determine free radical scavenging activity of flavonoids. It was found that the flavonol glycosides could markedly inhibit the luminescent signal, which indicated that they are mainly responsible for the antioxidant activities of G. biloba leaves. Total antioxidant activity of these flavonoids was used to evaluate the differences of G. biloba leaves collected in 13 habitats. The combination of chemical and activity analysis can provide a valid method to quantify the bioactive components in G. biloba leaves, and this may be a more rational approach to the quality assessment of G. biloba leaves.

[Chemical constituents from Polygonum capitatum and their antioxidation activities in vitro].

OBJECTIVE: To study the chemical constituents from Polygonum capitatum and their antioxidation activties in vitro. METHODS: Compounds were isolated and purified by silica, polyamide and Sephadex LH-20 column chromatography. Their structures were identified by physicochemical properties and spectral analyses. The isolated compounds were screened with three antioxidation models in vitro. RESULTS: Ten compounds were isolated and identified as galic acid (1), protocatechuic acid (2), progallin A (3), nimbecetin (4), quercetin (5), quercitroside (6), tetracosylferulate (7), quercetin-3-O-beta-D-glucoside (8), beta-deucosterol and beta-sitosterol (10). Compounds 1-6 from this plant showed antioxidation activities. CONCLUSION: Compound 3 and 7 are isolated from this genus for the first time, compound 2, 4 and 9 are isolated from this plant for the first time, the pharmacological activities of the pure compounds from this plant are also reported firstly.