Objective To compare the effect of Compound Wurenchun Capsule (CWC) and Wuzhi Capsule (WZC), CWC single and long-term administration on the pharmacokinetics of tacrolimus (FK506). Methods Twenty-four rats were randomly divided into FK506, CWC + FK506, WZC + FK506 and CWC7d + FK506 groups, with six rats in each group. Rats in FK506, CWC + FK506, and WZC + FK506 groups were given a single gavage with FK506, CWC + FK506, and WZC + FK506 respectively. Rats in CWC7d + FK506 group was given a multiple gavage regimen of daily CWC gavage for 6 d, CWC and FK506 on day 7. Blood sample from orbit before and after gavage at different time points (CWC7d + FK506 group before and after the last administration) were tested for FK506 blood concentration and the pharmacokinetic parameters were calculated. Results Compared with FK506 group, peak blood concentration (Cmax) and area under the curve (AUC0-t) of FK506 increased significantly (P < 0.05, 0.01), body retention time (MRT0-t) of FK506 prolonged significantly (P < 0.05, 0.01), the apparent volume of distribution (V/F) and the drug elimination rate (CL/F) of FK506 decreased significantly (P < 0.01) in WZC + FK506 and CWC + FK506 groups. Compared with WZC + FK506 group, AUC0-t of FK506 increased significantly (P < 0.01), CL/F of FK506 decreased significantly (P < 0.01) in CWC + FK506 group. Compared with CWC + FK506 group, Cmax of FK506 increased significantly (P < 0.01), time to peak blood concentration (tmax) of FK506 shortened significantly (P < 0.05), AUC0-t of FK506 increased significantly (P < 0.05), CL/F of FK506 decreased significantly (P < 0.05) in CWC7d + FK506 group. Conclusion Both CWC and WZC can increase Cmax and AUC0-t, prolong MRT0-t, reduce V/F and CL/F of FK506. CWC is better than WZC in increasing AUC0-t and inhibiting CL/F of FK506. CWC long-term administration is better than single-dose in improving Cmax, AUC0-t and reducing tmax of FK506.

Objective: To optimize the extract process of anthraquinones from the Rhei Radix et Rhizoma (RRR). Methods: The contents of eight bound anthraquinones (Bas) and five free anthraquinones (Fas), a total of 13 original anthraquinones, and total anthraquinones (Tas) in RRR were determined by HPLC. L9(34) orthogonal table was used, with ethanol concentration, solvent ratio, extract time, and number of extract as factors and extract quantity of Tas, five Fas, and eight Bas as the investigation index, respectively, the extract processes of Tas, Fas, and Bas from RRR were optimized. The optimized processes were verified by magnification test. Results: The optimum extract process of Tas and Fas was as follows: five times of 75% ethanol, extracting five times by reflux, 30 min for each time. Using this process, the extraction rate for both original anthraquinones and Tas was about 90%, and Fas extraction rate was over 160%. The optimum extract process of Bas was as follows: five times of 95% ethanol, extracting three times by reflux, 60 min for each time. Using this process, original anthraquinones extraction rate was close to 90%, and the one of Bas was over 80%. Conclusion: The optimized process is simple, stable, feasible, and repeatable, and can be used for the extraction of Tas, Fas, and Bas from RRR, respectively.

OBJECTIVE: To establish the fingerprint of the serum containing drug of Yinchenhao Decoction (YCHD), and analyze the drug-originated constituents in serum. METHODS: An HPLC method was set up on a Symmetry C18(4.6 mm × 250 mm, 5 μm) column with a Security Guard Cartridges C18 (4.0 mm × 3.0 mm) guard column by gradient elution of acetonitrile-0.1% phosphoric acid in water at the flow rate of 1.0 mL · min-1. The column temperature was maintained at 30℃. The detection was carried out at 238 and 440 nm. SD rats were taken as serum donors. Ten batches of serum containing drugs were prepared after ig administration of YCHD. The fingerprints of these serum samples were determined, the common modes were established, and the similarities between fingerprints were evaluated. Drug-originated constituents in blood were distinguished by comparing the fingerprint of serum containing YCHD with the one of control serum. The sources of drug-originated constituents were analyzed by comparing the fingerprint of serum containing YCHD with the ones of serum containing individual herbs of YCHD. Drug-originated constituents in blood were identified by comparing the retention time and UV spectra of peaks in fingerprint of serum containing YCHD with the ones of YCHD and reference substances, respectively. RESULTS: The similarities between the fingerprints of ten batches of serum containing YCHD and common models at 238 and 440 nm were greater than 0.904 and 0.903, respectively. Twenty common peaks of drug-originated constituents were identified in the fingerprints of serum containing drug of YCHD at 238 and 440 nm. Out of them, six were the original compounds, the other 14 were metabolites. Three constituents out of six original form ones were identified as geniposidic acid and genipo-side (iridoids originated from GF), and rhein (anthraquinone from RRR), and the other three were identified as an anthraquinone from RRR and two crocetin derivatives from FG. Out of 14 metabolites, one was identified as the metabolite of iridoid, four were from anthraquinone, and nine were from crocetin derivatives. CONCLUSION: The established method is accurate, reliable and can be used for the analysis of drug-originated constituents in serum containing drug of YCHD.

OBJECTIVE: To establish a quality control method of Rhei Radix et Rhizome (RRR) by fingerprint and simultaneous determining eight components by quantitative analysis of multi-components by single marker (QAMS). METHODS: An HPLC method was set up. Symmetry C18 column (4.6 mm×250 mm, 5 μm) was used. Methanol-0.1% phosphoric acid was used as gradient mobile phase. The flow rate was 1.0 mL·min^-1. The column temperature was 30℃ and detection wavelength set at 254 nm. Eleven batches of RRR were determined and a common mode of fingerprint maintained at established. A method was developed for QAMS to determine gallic acid, catechin, emodin-8-glucoside, aloe-emodin, rhein, emodin, chrysophanol, and physcion in RRR. Rhein was selected as internal reference: the relative correction factors (RCF) of other seven components to rhein were calculated. The contents of the eight components in ten batches of RRR were determined by both external standard method and QAMS. The QAMS method was evaluated by comparison of its assay results with that of external standard method. RESULTS: There were 35 common peaks in the fingerprints often batches RRR, eight of them were identified as gallic acid, catechin, emodin-8-glucoside, aloe-emodin, rhein, emodin, chrysophanol, and physcion. These eight components showed good linear relationship in the range of 0.0624-1.56 μg (r=0.9995), 0.18-4.5 μg (r=0.9998), 0.0288-0.72 μg (r=0.999 9), 0.0198-0.495 μg (r=0.9999), 0.0505-1.2625 μg (r=0.9999), 0.0637-1.5925 μg (r=0.999 9), 0.098-2.45 μg (r=0.9999), and 0.163-4.075 μg (r=0.9999), respectively. The established RCF had good reproducibility. No significant differences were found between the quantitative results of external standard method and QAMS. CONCLUSION: The developed method is accurate, feasible, and can be used for the overall quality control of RRR.

OBJECTIVE: To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneous determining six lignanoids in Schisandra chinens(S. chinensis). METHODS: A HPLC method was set up. Lichrosphere C18 column(4.6 mm × 250 mm, 5 μn) was used. Acetonitrile-water was used as gradient mobile phase. The flow rate was 1.0 mL · min-1. The column temperature was 30°C and detection wavelength was 217 nm. A method was developed for QAMS to determine schizandrol A, schizandrol B, schisantherin A, deoxyschizandrin, schizandrin B, and schizandrin C in 5. chinensis. Schizandrol A was selected as internal standard; the relative correction factors (RCF) of schizandrol B, schisantherin A, deoxyschizandrin, schizandrin B, and schizandrin C to schizandrol A were calculated. The contents of the six lignanoids in 12 different batches of S. chinensis were determined by both external standard method and QAMS. The QAMS method was evaluated by comparison of its assay results with that of external standard method. RESULTS: The linear range of schizandrol A, schizandrol B, schisantherin A, deoxyschizandrin, schizandrin B, and schizandrin C were within 0.09-1.62 μg(r=0.9999), 0.0408-0.7344 μg (r=0.9999), 0.0208-0.3744 μg(r=0.9999), 0.0242-0.4356 μg(r=0.9999), 0.0532-0.9576 μg(r=0.9999) and 0.0258-0.4644 μg(r=0.9999), respectively. No significant differences were found between the quantitative results of external standard method and QAMS. CONCLUSION: The developed method is accurate, feasible, and can be used for quality evaluation of S. chinensis.

Objective: To research the influence of prescription compatibility on the extraction rate of main constituents in Yinchenhao Decoction (YCHD). Methods: The extraction volume of two iridoids (geniposide and deacetylasperulosidic acid methyl ester), two crocetin derivatives (crocin I and crocin II), three bound anthraquinones (aloe-emodin-8-O-glucoside, chrysophanol-1-glucoside, and emodin-8-glucoside), five free anthraquinones (aloe-emodin, rhein, emodin, chrysophanol, and physcion), two tannins (gallic acid and catechin), and chlorogenic acid in Artemisiae Scopariae Herba (ASH), Gardeniae Fructus (GF), Rhei Radix et Rhizoma (RRR), the compatibility of ASH and GF (compatibility A), the compatibility of ASH and RRR (compatibility B), the compatibility of GF and RRR (compatibility C), and the compatibility of ASH, GF, and RRR (compatibility D) were determined by HPLC. Taking the extraction volume in single medicine as 100%, the extraction rates of 15 constituents mentioned above in compatibilities A-D were calculated. Results: The extraction rates of geniposide, crocin I, and crocin II in compatibilities A, C, and D, deacetylasperulosidic acid methyl ester in compatibility C, gallic acid in compatibilities C and D, and chlorogenic acid in compatibilities A-D decreased. The extraction rates of deacetylasperulosidic acid methyl ester in compatibilities A and D, three bound anthraquinones, aloe-emodin, and chrysophanol in compatibility B, chrysophanol and physcion in compatibilities C and D increased. Conclusion: Prescription compatibility has some influence on the extraction rate of main constituents in YCHD.

OBJECTIVE: To establish the fingerprint of the serum containing rhubarb bound anthraquinones, and to analyze the drug-originated constituents in serum containing drugs by serum pharmacochemistry. METHODS: Rat serum containing drugs were prepared after intragastric administration of the extract of rhubarb bound anthraquinones. Fingerprints of 10 batches serum containing drugs were determined by HPLC, and the common mode was established. Comparing the chromatogram of serum containing drugs with the one of blank serum, the common peaks of drug-originated constituents in blood were identified. Comparing the retention time and spectrum of the chromatographic peaks of serum containing drugs with the ones of extract, the sources of drug-originated constituents were analyzed. Comparing the retention time and spectrum of the chromatographic peaks of serum containing drugs with the ones of mixed reference substances, some drug-originated constituents were identified. RESULTS: There are 20 common peaks of drug-originated constituents in 10 batches serum containing drugs, 14 of them were in vitro prototype, and the other 6 were metabolites in vivo. Thirteen out of 14 prototype constituents were identified as aloe emodin-8-O-glucopyranoside, rhein-8-O-glucopyranoside, emodin-1-O-glucopyranoside, chrysophanol-1-O-glucopyranoside, chrysophanol-8-O-glucopyranoside, emodin-8-O-glucopyranoside, aloe emodin-3-CH2-O-glucopyranoside, physcion-8-O-glucopyranoside, aloe emodin, rhein, emodin, chrysophanol, and physcion, respectively. Other 1 prototype constituent and 6 metabolites possessed the same UV absorption character as that of anthraquinones. CONCLUSION: The method established in this study is accurate and feasible, and can be used for the analysis of the drug-originated constituents in serum containing rhubarb bound anthraquinones.

OBJECTIVETo study the differences in the toxicity of vinegar-processed Kansui Radix on normal and cancerous ascites model rats.

METHODNormal and cancerous ascites model rats were taken as the research objects and orally administered with different doses of vinegar-processed Kansui Radix for 7 d. Pathological sections were prepared to observe the damages in liver, stomach, intestinal tissues in rats and detect the impacts on serum, liver, stomach and intestinal tissues and the oxidative damage index.

RESULTCompared with the blank group, all of normal administration groups and model groups showed significant damages in liver, stomach and intestinal tissues. Compared with the model groups, all of normal administration groups revealed notable alleviation in damages. Compared with the blank group, the model groups showed significant increases in AST, ALT and MDA in serum and liver (P < 0.01) and a significant decrease in GSH in serum and liver, stomach, intestinal tissues (P < 0.01). Compared with the blank group, the results showed significant decreases in ALT, AST in serum and ALT in liver in model low, medium and high dose groups and AST activity in liver tissues in the normal high dose group (P < 0.05, P < 0.01); significant decreases in GSH in serum and stomach tissues in normal low, medium and high dose groups and GSH content in liver and intestinal tissues in normal medium and high dose groups (P < 0.05, P < 0.01); notable rises in MDA in liver tissues in normal low, medium and high dose groups and MDA content in serum and stomach and intestinal tissues in normal medium and high dose groups (P < 0.05, P < 0.01). Compared with model groups, data revealed significant decreases in ALT, AST in serum in model low, medium and high dose groups, AST in liver tissues of model medium and high dose groups and ALT activity in liver in the model high dose group (P < 0.05, P < 0.01); significant increases in GSH content in serum and stomach tissues of model low, medium and high dose groups, GSH in liver tissues in model medium and high dose groups and GSH in intestinal tissues in the high dose groups (P < 0.05, P < 0.01); and notable declines in MDA content in serum in model low, medium and high dose groups, MDA in liver tissues of model medium and high dose groups and MDA in stomach and intestinal tissues the high dose group (P < 0.05, P < 0.01).

CONCLUSIONAccording to the study, vinegar-processed Kansui Radix showed a significant lower toxicity liver, stomach, and intestines of cancerous ascites model rats, which provided a basis for clinical safe application of vinegar-processed Kansui Radix based on symptom-based prescription theory.

Acetic Acid ; chemistry ; Animals ; Chemistry, Pharmaceutical ; methods ; Drug Prescriptions ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; toxicity ; Euphorbia ; chemistry ; toxicity ; Intestines ; drug effects ; pathology ; Liver ; drug effects ; metabolism ; pathology ; Male ; Neoplasms ; drug therapy ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Plant Roots ; chemistry ; toxicity ; Rats ; Rats, Sprague-Dawley

To study the function of expelling water retention with drastic purgative of different polarities of Kansui Radix stir-baked with vinegar on the cancerous ascites model rats, the furosemide was taken as positive control drug, and the cancerous ascites model rats were respectively orally administered with different polarities of Kansui Radix stir-baked with vinegar for 7 d. The amount of urine and ascites, the level of urinary sodium, potassium, chloride ion and pH, and the content of PRL1, AII, ALD in serum were investigated. Compared with model groups, ethyl acetate extract group showed a decreasing trend in ascites; the amount of urine of showed a significant increase (P < 0.05); the level of urinary sodium, potassium, chloride ion (P < 0.05, P < 0.01), pH (P < 0.05), and the content of PRL1, AII, ALD in serum all showed a significant decrease (P < 0.01). The effects of petroleum ether extract and n-butanol extract were weaker than that of ethyl acetate extract. The water exact was the weakest. The results showed that ethyl acetate extract is the active part of Kansui Radix stir-baked with vinegar on the function of expelling water retention with drastic purgative on the cancerous ascites model rats, alleviating the water-electrolyte disorder and body fluid acid-base imbalance, regulating the renin angiotensin aldosterone system.
Animals ; Ascites ; drug therapy ; metabolism ; Cathartics ; administration & dosage ; chemistry ; isolation & purification ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Euphorbia ; chemistry ; Humans ; Male ; Plant Roots ; chemistry ; Potassium ; urine ; Rats ; Rats, Sprague-Dawley ; Sodium ; urine ; Water ; metabolism

OBJECTIVETo investigate the effect of 6-gingerol, the main active component of ginger, on hair shaft elongation in vitro and hair growth in vivo.

METHODSFirstly, Hair follicles were co-cultured with 3 different concentration of 6-gingerol for 5 days and hair elongation in three groups was measured. Secondly, The proliferative effect of 6-gingerol on DPCs was measured using MTT assay. Thirdly, the expression of Bcl-2 and Bax in DPCs were measured using Western blotting. In vivo study, the influence of 6-gingerol on hair growth in C57BL/6 rats was measured through topical application of 6-gingerol on the dorsal skin of each animal.

RESULTSThe length of hair shaft in 20 microg/ml 6-Gingerol group (0.50 +/- 0.08 mm) is less than 0 microg/ml (0.66 +/- 0.19) mm and 10 microg/ml (0.64 +/- 0.03) mm 6-Gingerol group (P < 0.05). In cell culture, compared to 0 microg/ml and 5 microg/ml 6-Gingerol, 10 microg/ml 6-Gingerol can significantly inhibited the proliferation of DPCs (P < 0.05). Along with the growth inhibition of DPCs by 6-gingerol, the Bax/Bcl-2 ratio increased obviously. In vivo study, the hair length and density decreased a lot after using 1 mg/ml 6-gingerol.

CONCLUSIONS6-Gingerol can suppress human hair shaft elongation because it has pro-apoptotic effects on DPCs via increasing Bax/Bcl-2 ratio. It might inhibit hair growth by prolonging the telogen stage in vivo.

Animals ; Catechols ; pharmacology ; Cell Culture Techniques ; Cells, Cultured ; Fatty Alcohols ; pharmacology ; Hair ; drug effects ; growth & development ; Hair Follicle ; drug effects ; growth & development ; Humans ; Mice ; Mice, Inbred C57BL ; Plant Extracts ; pharmacology ; Rats ; bcl-2-Associated X Protein ; metabolism