Danggui Buxue Decoction Research Articles

Astragaloside IV liposomes ameliorates adriamycin-induced nephritic syndrome in rats

Background : Radix Astragali was one of the main compositions of ‘Modified Danggui Buxue Decoction’ used for treatment of various kidney diseases. Astragaloside IV was the active composition of Radix Astragali . Astragaloside IV liposomes were used for the treatment of adriamycin-induced nephropathy (AN) rats. The aim of the study was to study the effect of astragaloside IV liposomes on AN rats, and to test through regulating the expression of nephrin, integrins and integrin-linked kinase (ILK) in renal tissues. Methods : The rats were given a single tail intravenous injection of adriamycin (6 mg/kg) within 1 week, and then divided into four groups including normal, model, benazepril and astragaloside IV liposomes group. They were all orally administered dosage of benazepril and astragaloside IV liposomes once daily for 8 weeks. Results : Astragaloside IV liposomes significantly reduced the proteinuria of AN rats at 28, 42 and 56 days. astragaloside IV liposomes could increase the mRNA and protein expression of nephrin, integrin α3, down-regulate the expression of ILK to alleviate the podocyte damage and restore glomerular selective filtration function. Conclusions : Astragaloside IV liposomes could enhance renal function and protect podocyte to ameliorate the adriamycin-induced nephritic syndrome. Key words ：Astragaloside IV, Liposomes, Ardiamycin-induce nephropathy, Podocyte

ASTRAGALOSIDE IV LIPOSOMES AMELIORATES ADRIAMYCIN-INDUCED NEPHRITIC SYNDROME IN RATS MING-GANG

Background: Radix Astragali was one of the main composition of ‘Modified Danggui Buxue Decoction’ used for treatment of various kidney
 diseases. Astragaloside IV was the active composition of Radix Astragali. Astragaloside IV liposomes were used for the treatment of
 adriamycin-induced nephropathy (AN) rats. The aim of the study was to study the effect of astragaloside IV liposomes on AN rats, and to test
 through regulating the expression of nephrin, integrins and integrin-linked kinase (ILK) in renal tissues.
 Methods: The rats were given a single tail intravenous injection of adriamycin (6 mg/kg) within 1 weeks, and then divided into four groups
 including normal, model, benazepril and astragaloside IV liposomes group. They were all orally administered dosage of benazepril and
 astragaloside IV liposomes once daily for 8 weeks.
 Results: Astragaloside IV liposomes significantly reduced the proteinuria of AN rats at 28, 42 and 56 days. astragaloside IV liposomes could
 increase the mRNA and protein expression of nephrin, integrin α3, down-regulate the expression of ILK to alleviate the podocyte damage and
 restore glomerular selective filtration function.
 Conclusions: Astragaloside IV liposomes could enhance renal function and protect podocyte to ameliorate the adriamycin-induced nephritic
 syndrome.

Pharmacokinetic comparison of seven major bio-active components in normal and blood deficiency rats after oral administration of Danggui Buxue decoction by UPLC-TQ/MS

Blood deficiency is commonly encountered among women, and is the root of many gynecological disorders. Danggui Buxue Decoction (DBD), a classical traditional Chinese formula which is composed of Astragali Radix (AR) and Angelicae Sinensis Radix (ASR) at the ratio of 5:1 (w/w), is widely used in TCM clinics for treatment of blood deficiency syndrome. This study is to compare the in vivo pharmacokinetic properties of seven major bio-active components in normal and blood deficiency rats after oral administration of DBD. Blood deficiency rats were induced by bleeding from orbit at the dosages of 5.0mL/kg each day for 12 days. Normal and blood deficiency rats were administrated of DBD on the 12th day at the dosage of 20g/kg, and blood was collected at different time points after then. Concentrations of ferulic acid, caffeic acid, butylphthalide, ligustilide, calycosin-7-O-β-glucoside, ononin, and astragaloside IV in plasma were quantified by UPLC-TQ/MS, and the main pharmacokinetic parameters were calculated by DAS 2.0. It was found that Cmax, Tmax and MRT0~T of astragaloside IV, Cmax, T1/2Z, AUC0~T and MRT0~T of calycosin-7-O-β-glucoside, T1/2Z and AUC0~T of ferulic acid, T1/2Z, AUC0~T and MRT0~T of ononin, and MRT0~T of ligustilide, butylphthalide, and caffeic acid in blood deficiency rats was significantly different (P<0.05) from normal rats. This study was the first report about pharmacokinetic investigation in blood deficiency animals which was conducted by bleeding. And the results demonstrated that the seven DBD constituents in normal and blood deficiency rats had obvious differences in some pharmacokinetic characteristics, suggesting that the rate and extent of drug metabolism were altered in blood deficiency animals.

Comparative tissue distribution profiles of five major bio-active components in normal and blood deficiency rats after oral administration of Danggui Buxue Decoction by UPLC-TQ/MS

Astragali Radix (AR) and Angelicae Sinensis Radix (ASR) were frequently combined and used in China as herbal pair called as Danggui Buxue Decoction (DBD) for treatment of blood deficiency syndrome, such as women's ailments. This study is to investigate the tissue distribution profiles of five major bio-active constituents (ferulic acid, caffeic acid, calycosin-7-O-β-glucoside, ononin and astragaloside IV) in DBD after oral administration of DBD in blood deficiency rats, and to compare the difference between normal and blood deficiency rats. The blood deficiency rats were induced by bleeding from orbit at the dosages of 5.0mLkg−1 every day, and the experimental period was 12 days. At the finally day of experimental period, both normal and blood deficiency rats were orally administrated with DBD, and then the tissues samples were collected at different time points. Ferulic acid, caffeic acid, calycosin-7-O-β-glucoside, ononin and astragaloside IV in different tissues were detected simultaneously by UPLC-TQ/MS, and the histograms were drawn. The results showed that the overall trend was CLiver>CKidney>CHeart>CSpleen>CLung, CC-30min>CM-30min>CM-60min>CC-5min>CM-5min>CC-60min>CM-240min>CC-240min. The contents of the detected compounds in liver were more than that in other tissues no matter in normal or blood deficiency rats. Compared to normal rats, partial contents of the compounds in blood deficiency rats’ tissues at different time points had significant difference (P<0.05). This study was the first report about tissue distribution investigation in blood deficiency animals which is conducted by bleeding. And the results demonstrated that the five DBD components in normal and blood deficiency rats had obvious differences in some organs and time points, suggesting that the blood flow and perfusion rate of the organ were altered in blood deficiency animals.

Protection of ultra-filtration extract from Danggui Buxue Decoction (当归补血汤) on oxidative damage in cardiomyocytes of neonatal rats and its mechanism

To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction (EDBD) was able to protect cardiomyocytes from oxidative injury of rats induced by hydrogen peroxide (H(2)O(2)) and its potential mechanism. Myocardial cells from 1- to 2-day-old neonatal rats were cultured in Dulbecco's modified Eagle's medium low-glucose and Ham's F12 medium (1:1), and the cellular injury was induced by H(2)O(2). The ultra-filtration extract mixture from Angelica sinensis and Hedysarum polybotrys was given in three doses of 3.75, 7.5, and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase (LDH) and creatine kinase (CK) releases in the medium, superoxide dismutase (SOD) activities, and intracellular malondialdehyde (MDA) and myeloperoxidase (MPO) contents. The levels of caspase-3 and heat shock protein 70 (hsp70) mRNA expression in cardiomyocytes were measured by reverse transcription polymerase chain reaction. The EDBD could protect the cardiomyocytes from H(2)O(2) injury in a dosedependent manner (3.75, 7.50, and 15.00 mg/mL). The EDBD could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents, increase SOD activity, up-regulate hsp70 expression, and down-regulate caspase-3 expression. The EDBD has protection on cardiomyocytes injured by H(2)O(2) through improving cell antioxidant ability, up-regulating hsp70 expression, and inhibiting caspase-3 activity.

Antifibrotic effect of the Chinese herbs Modified Danggui Buxue Decoction (加味当归补血汤) on adriamycin-induced nephropathy in rats

To investigate the antifibrotic effect of the Chinese herbs Modified Danggui Buxue Decoction (, MDBD) on adraimycin-induced nephropathy in rats. Thirty-two male Sprague Dawley albino rats were randomly divided into 4 groups: the control, model, and two treatment groups, with 8 in each group. Nephropathy was induced in the latter 3 groups by intravenous injection of adriamycin. Rats in the two treatment groups received intragastric administration of benazepri (a positive control) or MDBD, which is composed of extracts of Radix Angelicae sinensis, Astragalus membranaceus (Fisch.) Bge and Rhizoma chuanxiong. Serum albumin, blood lipids, 24-h urine protein and urine N-acetyl-b-D-glucosaminidase (NAG) were measured every 2 weeks. The ratio of kidney to body weight was measured. The expressions of extracellular matrix proteins in the renal cortex, including colleagen IV (Col-IV) and fibronectin (FN), were examined by immunohistochemistry, and the transcription of genes encoding transforming growth factor β1 (TGF-β1), the tissue inhibitors of matrix metalloproteinase 1 (TIMP-1) and matrix metalloproteinase 9 (MMP-9) were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) at the end of the 8-week treatment. Compared with the untreated rats in the model group, MDBD significantly increased serum albumin, lowered the blood lipids and decreased the ratio of kidney to body weight. MDBD significantly reduced the excretion levels of urinary protein and NAG as well as the accumulation of extracellular matrix (ECM), including Col-IV and FN, in the renal cortex. Further, MDBD decreased TIMP-1 and TGF-β1 gene expressions and increased MMP-9 gene expression in the kidney. MDBD was effective in treating the rat model of nephropathy. The clinical benefit was associated with reduction of renal fibrosis. The antifibrotic effect of MDBD may be mediated through the regulation of TIMP-1, MMP and TGF-β1 gene expressions.

Correlation of powder properties and formability of traditional Chinese medicine pellets

To investigate the correlation of powder properties and formability of traditional Chinese medicine pellets prepared by extrusion/spheronization. The present effort was designed to evaluate the effects of the characters of powder on pellets formation by extrusion-spheronization, taking Danggui Buxue decoction extracts as a model drug and microcrystalline cellulose (MCC) as an ideal balling material for extrusion-spheronization. The diameter of powder had correlation with the plane critical angle of pellets in a certain scope and the sticky coefficient of wet mass affected roundness of pellets. Properties of powder have influence on formability of traditional Chinese medicine pellets.

Concentration of Components in Danggui-Buxue Decoction by Macroporous Resin Chromatography Guided with LC

A method for simultaneous concentration of multiple bioactive components (MBCs) in Danggui-Buxue Decoction (DBD) was comprehensively investigated in this work. The column chromatography with macroporous resin ADS-7 as an absorbent was used for concentration of the MBCs in DBD under the guidance of liquid chromatography (LC). After one-run treatment with the resin, the contents of major flavonoids reached 18.3 and 13.9% in the eluting fraction of 50 and 70% ethanol, which were 177.2 and 134.6 times higher than those in DBD crude extract. The contents of main saponins reached 8.7 and 9.9% in the eluting fractions of 10 and 30% ethanol, which were 32.8 and 37.3 times higher than those in DBD crude extract; and ferulic acid and ligustilide were also enriched in the eluting fractions of 10 and 70% ethanol.

Researches on relationships between main active ingredients and physical and chemical parameters of Dangguibuxue decoction

To study relationships between the main active ingredients and physical and chemical parameters of dangguibuxue decoction, and to explore the feasibility of characterization the quality of decoction by physical and chemical parameters. Solutions of ferulic acid and astragaloside with different concentrations were prepared, of which physical and chemical parameters such as surface tension, pH value and conductivity, etc were determined at 25 degrees C. And polynomial regression analysis was used to analyze these data. With the increase of concentration of ferulic acid, pH value was decreased and conductivity was increased. And surface tension was decreased in the aqueous solution of astragaloside. In the combined solution of ferulic acid and astragaloside, ferulic acid was the main influencing pH value and conductivity, while astragaloside was the main factor influencing surface tension. On one hand, astragaloside concentration has a significant correlation with surface tension, but almost no correlation with pH value and conductivity. On the other hand, ferulic acid concentration has a strong correlation with pH value and conductivity, but almost no correlation with surface tension. In addition, each parameter has strong anti-interference ability and almost didn't change with the addition of another material. These preliminary results suggested that such characteristic parameters could be used to characterize the decoction quality.

Chemical composition and inhibitory effect on hepatic fibrosis of Danggui Buxue Decoction

The present study was carried out to investigate chemical components and therapeutic effects on hepatic fibrosis of Danggui Buxue Decoction (DBD) in rabbits. The chemical components of DBD were analyzed by LC-DAD-MS. Schistosomiasis hepatic fibrosis was employed as models of hepatic fibrosis. The rabbits were divided into treatment and control groups. DBD was orally administered at a dose of 50 g/kg rabbit's weight in treatment group. We evaluated the therapeutic effects of DBD on hepatic fibrosis in rabbits using B-ultrasound images, histopathologic examination, and hyaluronic acid content in serum. The results indicated that DBD had good therapeutic effects for hepatic fibrosis.

Effects of Danggui Buxue Decoction (当归补血汤) on lipid peroxidation and MMP-2/9 activities of fibrotic liver in rats

To explore the mechanism of Danggui Buxue Decoction (, DBD) on the liver fifibrosis related to hepatic lipid peroxidation and matrix metalloproteinases (MMP) -2/9 activities. The liver fifibrosis in 28 rats was induced by an injection of carbon tetrachloride (CCl(4)) and fed with high lipid and low protein diet for 6 weeks, the model rats were randomly divided into the model group and DBD treated group, 14 in each group, and another 10 rats as the normal group were observed as well. Rats in the DBD group were administered with DBD at the dose of 6 g/kg body weight for 6 weeks since CCl(4) intoxication. The hepatic inflammation and fibrosis were examined with HE and Sirius red stain. The liver function including serum alanine aminotransamine (ALT), aspartate transamine (AST), albumin (Alb) and total bilirubin (TBIL), liver triglyceride (TG) and malondialdehyde (MDA) contents, superoxide dismutase (SOD) activity were assayed. Hepatic hydroxyproline (Hyp) content was detected with Jamall's method. The alpha-SMA expression was analyzed by immunohistochemistry and the Western blot. Liver MMP-2 mRNA was analyzed with Real-time PCR, and MMP-2/9 activities were measured with gelatin zymography and in situ zymography. Compared with the normal group, the levels of ALT, AST and TBIL, the content of Hyp, TG and MDA were remarkably increased, the Alb content and SOD activity were signifificantly decreased in the model group (P<0.05), and higher levels of MMP-2 mRNA and MMP-2/9 activities (P<0.01), the hepatic fatty degeneration and collagen accumulation and fifibrosis at liver were observed. Compared with the model control, DBD group showed slighter hepatic fatty degeneration and collagen deposition, and had lower levels of ALT, AST and TBIL activities, lower contents of MDA, TG and Hyp, but higher SOD level and Alb content (P<0.05), and DBD also down-regulated MMP-2 mRNA expression and decreased MMP-2/9 activities in the fifibrotic livers (P<0.01). The action of DBD against liver fibrosis is related to prevent lipid peroxidation and inhibit MMP-2/9 activities in the fibrotic livers.

Efficacy of auxiliary therapy with Danggui Buxue Decoction No.1 (当归补血汤1号) in treating patients of non-small cell lung cancer at peri-operational stage

To observe the therapeutic efficacy of Danggui Buxue Decoction No.1 in treating patients of non-small cell lung cancer (NSCLC) at the peri-operational stage and its impact on the patients' immune function. Eighty-two NSCLC patients were randomly assigned to two groups equally, the control group and the test group, they were given conventional treatment, while to the test group, DB1 were given additionally. The observation was conducted by testing the changes of T-lymphocyte subsets, natural killer (NK) cell activity, serum levels of immunoglobulin (IgA, IgM, IgG), interleukin-2 (IL-2), tumor necrosis factor alpha (TNF-alpha), cytokeratin fragment 19 (CYFRA21-1) and carcinoembryonic antigen (CEA) in NSCLC patients before and after administration of DB1, and analyzing the patients' general condition. The level of CD3(+), CD4(+), the ratio of CD4 and CD8(+), IgA, IgM, IgG and IL-2 decreased in patients with NSCLC on day 1 after operation, and the level of CD8 and TNF-alpha increased compared to pre-operation. While the levels of CD3(+), CD4(+), CD4 /CD8(+), NK cell activity, serum IgA, IgM, IgG, IL-2 began to elevate, CD8 and TNF-alpha levels began to decline in patients administered with DB1 on day 3 after the operation, earlier than patients who did not use the decoction. The level of CYFRA21-1 and CEA, was immediately decreased after operation in both groups. Applying DB1 to NSCLC patients at an early post-operational stage could alleviate the impairment and accelerate the recovery of immune function of patients to enhance their immunity. DB1 also shows an anti-tumor action to a certain degree.

Synergic effects of Danggui Buxue Decoction in reducing toxicity of cytoxan in tumor-bearing mice

To study the inhibitory effect of Danggui Buxue Tang (DBT), a compound traditional Chinese herbal medicine for supplementing blood, on tumor growth in tumor-bearing mice after inoculation of EL-4 cells, and its immune mechanism as well as its synergic effect in reducing toxicity of cytoxan (CTX). Experiment was carried out in tumor-bearing mice after inoculation of EL-4 cells. The mice were randomly divided into four groups after 7 days of the inoculation: untreated group, DBT-treated group [24 g/(kg x d)], CTX-treated group [7.5 mg/(kg x d)] and DBT plus CTX-treated group, with another ten normal mice as control. Inhibitory rate of tumor growth, survival time, immune function and variability of blood cells were measured in the mice during the experiment. After treatment of relevant interventions for 15 days, the tumor in the DBT-treated group, CTX-treated group and DBT plus CTX-treated group grew slower than the untreated group (P<0.05). Murine survival time in the DBT-treated group, CTX-treated group and DBT plus CTX-treated group was lengthened as compared with that in the untreated group (P<0.05). Compared with the untreated group, all kinds of immune indexes in the DBT-treated group and DBT plus CTX-treated group were significantly improved (P<0.05), while the immune indexes in the CTX-treated group were decreased (P<0.05). Compared with the CTX-treated group, all kinds of immune indexes in the DBT plus CTX-treated group were significantly improved (P<0.05). DBT can enhance the immune function in tumor-bearing mice and the inhibitory effect of DBT on tumor growth is related to the enhanced immune response. DBT can also increase the therapeutic effects and reduce the side effects of CTX.

Screening and identification of potential bioactive components in a combined prescription of Danggui Buxue decoction using cell extraction coupled with high performance liquid chromatography

A cell extraction coupled with high-performance liquid chromatography (HPLC) analysis has been developed to screen the potential bioactive components in combined prescription of Danggui Buxue decoction. The method was validated by using HL-7702 cells, RAW 264.7 cells and Caco-2 cell extraction. According to the hypothesis that, when cells are incubated together with the extract of traditional Chinese medicines (TCMs), the potential bioactive compounds in the extract should selectively combine with the cells, cell-combining compounds were analyzed by HPLC-diode array-evaporative light scattering detectors. Their structures were elucidated in comparison with available reference compounds, and further confirmed by HPLC-electrospray ionization time-of-flight mass spectrometry with accurate mass measurement. The results demonstrated that nine compounds were combined with the HL-7702 cells, seven with RAW 264.7 cells, and thirteen with the Caco-2 cell line. In view of the two key steps of drugs action, absorption by intestinal epithelium cells and interaction with target cells, this rapid and reliable method could be utilized to predict the bioactive constituents in TCMs, and it was in agreement with the characteristics of combined prescriptions of TCMs as multi-components, multi-target sites and multi-channel actions.

Analysis of interaction property of bioactive components in Danggui Buxue Decoction with protein by microdialysis coupled with HPLC–DAD–MS

Interaction of a commonly used combined prescription of Danggui Buxue Decoction (CPDBD) with protein was studied by microdialysis coupled with HPLC–DAD–MS. Nine compounds in CPDBD were unequivocally identified by comparing with their t R, MS data and UV spectra with those of reference compounds, and simultaneously quantified. Microdialysis recoveries and binding degrees of 20 compounds in CPDBD with bovine serum albumin (BSA) were determined. Recoveries of microdialysis sampling ranged from 66.9 to 91.5% with RSD below 3.0%, and the binding degrees of those to BSA ranged from 6.3 to 59.8% (0.3 mM BSA) and from 6.9 to 86.6% (0.6 mM BSA). The results were determined at pH 7.4 and the influence of different pH value was investigated. Furthermore, the binding degrees of eight reference compounds were determined separately under the same conditions, indicating a significant effect of the interaction of compounds with each other on their binding degrees to BSA. By comparing their binding degrees with BSA with those of proven active compounds in CPDBD, i.e. chlorogenic acid ( 3), ferulic acid ( 6), ononin ( 12) and calycosin ( 16), other five compounds were found to possess potential activities, which were tentatively identified as calycosin-7- O-β- d-glucoside-6- O-malonate ( 9), senkyunolide I or H ( 10), formononetin-7- O-β- d-glucoside-6- O-malonate ( 17), and two unknown compounds.

Screening and analysis of the multiple absorbed bioactive components and metabolites of Dangguibuxue decoction by the metabolic fingerprinting technique and liquid chromatography/diode‐array detection mass spectrometry

Based on the metabolic fingerprinting technique and liquid chromatography/diode array detection mass spectrometry (LC/DAD-MS), a method for rapid screening and analysis of the multiple absorbed bioactive components and metabolites of an oral solution of Dangguibuxue decoction (ODD) in rabbit plasma after oral administration of ODD was developed. The results obtained from a comprehensive comparative analysis of the fingerprints of the ODD and its metabolic fingerprints in rabbit plasma indicated that 46 components in the ODD were absorbed into the rabbit's body. Of them, ten components were tentatively identified from their MS and UV spectra and retention behaviors by comparing the results with the reported literature. They were calycosin-7-O-beta-D-glycoside, (6aR,-11aR)-hydroxy-9,10-dimethoxypterocarpan-3-O-beta-D-glycoside, ononin, L-3-hydroxy-9,10-dimethoxypterocarpan, formononetin, (3R)-7,2'-dihydroxy-3',4'-dimethoxyisoflavan, sedanenolide, E-ligustilide, Z-ligustilide, and Z-butylidenephthalide. In addition, 21 components were only found in the metabolic fingerprints, which suggested that they might be metabolites of some components in the ODD. The findings demonstrated that the proposed method could be used to rapidly and simultaneously analyze and screen the multiple absorbed bioactive constituents and metabolites in a formula of traditional Chinese medicines (TCMs) by comparing and contrasting the chromatographic fingerprints with its metabolic fingerprints. This is very important not only for the pharmaceutical discovery process and the quality control of crude drugs, but also to explain the curative mechanism of TCMs.

Screening and identification of permeable components in a combined prescription of Danggui Buxue decoction using a liposome equilibrium dialysis system followed by HPLC and LC-MS

A new method, i.e., liposome equilibrium dialysis followed by HPLC and LC-MS analysis, has been developed for the screening of permeable components in combined prescriptions of Danggui Buxue decoction (CPDBD). Multiple permeable components were simultaneously predicted by comparison of chromatograms of CPDBD extract before and after interaction with liposome membranes. A diode-array detector (DAD) and an evaporative light scattering detector (ELSD) were used, and the permeable compounds were identified by comparison with the available reference compounds and confirmed by on-line LC-MS. About fifteen compounds in a CPDBD extract were found to interact with liposome membranes. They were identified as calycosin-7-O-beta-D-glucoside (1), senkyunolide I or H (2), ononin (3), (6alphaR,11alphaR)-9,10-dimethoxypterocarpan-3-O-beta-D-glucoside (4), (3R)-2'-hydroxy-3',4'-dimethoxyisoflavan-7-O-beta-D-glucoside (5), calycosin (6), astragaloside IV (7), isoastragaloside II (8), formononetin (9), (6alphaR, 11alphaR),-3-hydroxy-9,10-dimethoxypterocarpan (10), (3R)-7,2'-dihydroxy-3',4'-dimethoxyisoflavan (11), astragaloside I (12), isoastragaloside I (13), E-ligustilide (14), and Z-ligustilide (15), respectively. Among all permeable components, 1, 3, 6, and 9 (flavonoids), 2, 14, and 15 (phthalides), and 7 (saponins) have been considered as major bioactive components in CPDBD. Therefore, this new method appears useful as a first step in the screening of bioactive components in natural products including Traditional Chinese Medicines (TCMs).

Live cell extraction and HPLC–MS analysis for predicting bioactive components of traditional Chinese medicines

A novel strategy for predicting bioactive components in traditional Chinese medicines (TCM) using live cell extraction and high performance liquid chromatography–diode array detection–mass spectrometry (HPLC–DAD–MS) analysis was proposed. The hypothesis is that when cells are incubated together with the extract of TCM, the potential bioactive components in the TCM should selectively combine with the cells, and the relative concentrations of the cell-combining components in the suspension medium should decrease, while the cell-combining components would be detectable in the extract of denatured cells. The identities of the cell-combining components could be determined by HPLC–DAD–MS analysis. Using the proposed approach, the potential bioactive components of Danggui Buxue decoction, a commonly used TCM for anaemia, and its compositions, Radix Angelica Sinensis and Radix Astragli for endothelial cells, were investigated. Six compounds in the extract of Danggui Buxue decoction were detected as the components selectively combined with endothelial cells, among them two were contributed by Radix Angelica Sinensis, and four by Radix Astragli. The identities of four of the six potential bioactive compounds were elucidated as ononoside, calycosin, 3-butylphthalide and ligustilide by HPLC–DAD–MS analysis. The results indicate that the proposed approach may be applied to predict the bioactive candidates in TCM.

Separation of compounds interacting with liposome membrane in combined prescription of traditional Chinese medicines with immobilized liposome chromatography

Immobilized liposome chromatography (ILC), the stationary phase of which has been regarded as a mimic biomembranes system was used to separate and analyze compounds interacting with liposome membrane in Danggui Buxue decoction, a combined prescription of traditional Chinese medicines (CPTCMs), and its compositions Radix Astragli and Radix Angelica Sinensis. More than 10 main peaks in the extract of Danggui Buxue decoction were resolved on the ILC column, suggesting that more than 10 components in the prescription have significant retention on ILC column. Ligustilide, astragaloside IV and formononetin, three main bioactive ingredients in Danggui Buxue decoction, were found to have relatively significant, while ferulic acid, another bioactive ingredient in the prescription, relatively weak retention on ILC column. Effects of the eluent pH and amount of immobilized phosphatidylcholine (PC) on separation of interactional compounds in the extract of Danggui Buxue decoction were also investigated. It was found that these two factors strongly affected the retention of some interactional compounds. In addition, the fractions partitioned with different solvents from water extract of this combined prescription were evaluated with this ILC column system.