Revealing the anti-inflammatory ingredients in wine-processed Radix et Rhizoma Rhei using immobilized cysteinyl leukotriene receptor type 1 as the stationary phase.

Despite the tremendous progress of wine-processed Radix et Rhizoma Rhei (Jiudahuang, JDH) in removing toxic heat from the blood in the upper portion of the body for hundreds of years, the deep understanding of its functional material basis of the anti-inflammatory ingredients remains unclear due to the lack of high specific and efficient methods. Herein, taking Cysteinyl leukotriene receptor type 1(CysLT1R) as the target protein, we established a chromatographic method based on the immobilized CysLT1R using haloalkane dehalogenases (Halo) at the C-terminus of the receptor in one step. After careful characterization by X-ray photoelectronic spectroscopy, immune-fluorometric analysis, and chromatographic investigations, the immobilized receptor was used to screen the anti-inflammatory ingredients in JDH. Aloe-emodin, rhein, emodin, chrysophanol, and physcion were identified as the main anthraquinone exerting anti-inflammatory effects in the drug. The association constants for the five compounds to bind with the receptor were calculated as (0.30 ± 0.06)× 105, (0.35 ± 0.03)× 105, (0.46 ± 0.05)× 105, (1.05 ± 0.14)× 105, and (1.66 ± 0.17)× 105 M-1 by injection amount-dependent method. Meanwhile, hydrogen bonds were identified as the main driving force for the five compounds to bind with CysLT1R by molecular docking. Based on these results, we believe that the immobilized receptor chromatography preserves historic significance in revealing the functional material basis of the complex matrices.

Effect of Porcine Large Intestine-processed Dahuang （Radix et Rhizoma Rhei） on Defecation and Colon Pathological Changes in Constipation Model Mice / 中医杂志

ObjectiveTo investigate the effect of porcine large intestine-processed Dahuang （Radix et Rhizoma Rhei） on defecation in constipation model mice and the possible mechanism. MethodsFifty Kunming mice were randomized to blank group （n=10） and model group （n=40）. Loperamide suspension at the dose of 8 mg/（kg·d） was given by gavage for four consecutive days to establish a model of constipation. The 24 successfully modeled mice were randomly divided into model group， processed Dahuang group， lactulose group， raw Dahuang group， with six mice in each group. Moreover， six randomly selected mice were chosen as control group. Since the fifth day， 8 mg/（kg·d） of loperamide suspension by gavage was given to the model group， processed Dahuang group， raw Dahuang group， and lactulose group； two hours later， the processed and raw Dahuang groups were administered with 0.6 g/（kg·d） of processed and raw Dahuang suspension， respectively， while the lactulose group was given 0.6 g/（kg·d） of latulose suspension， and the blank group and the model group were given 0.2 ml/10 g of distilled water by gavage， all for four days. The general condition， body weight after the last gavage， number of fecal particles within six hours， fecal wet weight， fecal water content ratio， intestinal propulsion rate and colonic histology changes by HE staining of each group were detected. ResultsThe body weight of the mice in the raw Dahuang group was significantly lighter than that in the other groups （P＜0.05 or P＜0.01）. The number of fecal particles， fecal wet weight and intestinal propulsion rate of mice significantly decreased in the model group than in the blank group （P＜0.05 or P＜0.01）. Compared to those in the model group， the number of fecal particles and fecal wet weight in the processed Dahuang group， lactulose group and raw Dahuang group significantly increased， and the fecal water content ratio in the raw Dahuang group increased as well （P＜0.05 or P＜0.01）. Compared to those in the processed Dahuang group， the number of fecal particles and fecal wet weight in the raw Dahuang group decreased， while the fecal water content ratio increased （P＜0.05 or P＜0.01）， and the fecal water content ratio in the lactulose group increased significantly （P＜0.05）. The intestinal propulsion rate in the processed Dahuang group was higher than that in the model group， lactulose group and raw Dahuang group （P＜0.05 or P＜0.01）. Histopathological analysis showed that the colonic crypts and goblet cells in the blank group were normal and clear， and the colonic muscular layer was thicker. The colonic crypts of the mice in the model group were damaged， with reduced goblet cells to varying degrees and changed colonic muscularis. In the lactulose group and raw Dahuang group， part of the crypts were broken， and the goblet cells were damaged to varying degrees， while in the processed Dahuang group， still the colonic tissue structure of the mice was relatively clear， and the colonic crypts and goblet cells were relatively normal， with thickened muscular layer of the colon. ConclusionPorcine large intestine-processed Dahuang could improve defecation in constipation model mice， and reduce the drastic purgation function of raw Dahuang， for which the mechanism may be related to the protection of colon histopathological damage.

Combination of comprehensive two-dimensional prostate cancer cell membrane chromatographic system and network pharmacology for characterizing membrane binding active components from Radix et Rhizoma Rhei and their targets.

Prostate cancer (PCa) is a common and fatal cancer for men and effective treatment options are still not enough for patients. Radix et Rhizoma Rhei had been applied to treat PCa long-termly and effectively when combined with surgical treatment and chemotherapy. However, its active components and target proteins are still not quite clear. As membrane receptors play a vital role in PCa, in this study, a novel strategy that combines comprehensive 2D 3-aminopropyltriethoxysilane-decorated prostate cancer cell (DU145) membrane chromatographic (CMC) system with network pharmacology approach was developed to characterize membrane binding active components proteins from Radix et Rhizoma Rhei and their targets. Thirteen active components were screened out by CMC system, among which emodin and rhapontigrnin with good membrane binding behaviors were validated to show ideal inhibitory effects on DU145 cells by cell viability and cell apoptosis assays. Five membrane proteins were predicted as the potential targets by the a specific network pharmacology approach, among which mast/stem cell growth factor receptor Kit (KIT) was identified as the most possible target by network data mining. Surface plasmon resonance analysis verified that the dissociation constant (KD) of rhapontigrnin and emodin with KIT was 6.06 × 10-5 M and 8.82 × 10-5 M, respectively. Our results showed that the combination of comprehensive 2D CMC system and network pharmacology based target identification could not only rapidly identify the membrane binding components but also find the potential membrane protein targets with high confidence, which could broaden the range of application scope of CMC, especially for the screening of active compounds from complex chemical samples using primary pathologic cell lines.

Quality Evaluation of Radix et Rhizoma Rhei Based on Gray Correlational Analysis and Functional Components / 世界科学技术-中医药现代化

Objective: To comprehensively evaluate the quality of Radix et Rhizoma Rhei based on gray correlationalanalysis and functional components, and to explore the difference of Radix et Rhizoma Rhei in different genuineproducing areas. Methods: HPLC was utilized to analyze 14 main compositions contained in the samples, includingemodin, rhein, chrysophanol, aloe-emodin, physcion, rheinoside, physcion glucoside, chrysophan, aloe-emodinglucoside, emodin methyl glycoside, sennoside, sennoside B, catechin and gallic acid. Then python 2.7 software wasemployed for gray correlation analysis of functional components closely related to the traditional efficacy of Radix et Rhizoma Rhei. Results: The qualities of Radix et Rhizoma Rhei grow in different areas were different. Tanggute Radix et Rhizoma Rhei grew in Tianzhu Gansu had the best effects of "expelling water retention and attacking the accumulation", and that grew in Yajiang Sichuan had the best effects of "clearing heat and removing toxin". Zhangye Radix et Rhizoma Rhei grew in Lixian Gansu had the best effect of"expelling stasis and unblocking the channels". Conclusion: Patternrecognition has broad prospects in the field of quality evaluation of traditional Chinese medicine. From the clinicalefficacy of traditional Chinese medicine, pattern recognition at the level of efficacy components can provide a new ideafor establishing a more complete and scientific quality evaluation system for traditional Chinese medicine.

Preventive Effect of Acupoint Application of Chinese Medicine on Post-stroke Constipation / 广州中医药大学学报

Objective To observe the effect of acupoint application of Chinese medicine on preventing constipation after stroke. Methods By means of randomized, double-blind, controlled study design, 96 patients of yang-syndrome stroke were randomly divided into observation group and control group, 48 cases in each group. Both groups were given conventional treatment for stroke and routine nursing for preventing constipation, and additionally, the control group was given acupoint application of placebo while the observation group was given acupoint application of self-made umbilical paste of Radix et Rhizoma Rhei(Dahuang) and Natrii Sulfas (Mangxiao). The incidence of constipation, frequency of laxative usage and adverse reactions of the two groups were observed. Results (1) The incidence of constipation of the observation group was lower than the control group on treatment day 7 and 14 , the difference being statistical significant(P 0.05). The observation group had better effect on improving stool properties(P 0.05). The adverse reaction of both groups was mild, and had no effect on the accomplishment of the treatment. Conclusion Acupoint application of Chinese medicine is effective and safe on preventing constipation after stroke, showing good prospects for clinical application.

Study on the extraction process of total anthraquinones in Radix et Rhizoma Rhei and their antilipemic effects.

BACKGROUND: Radix et Rhizoma Rhei has a gastric mucosal protective effect, major anti-gastritis and anti-peptic ulcer active constituents were emodin, aloe-emodin, chrysophanol, rhein, etc. The objective of the study was the extraction process of total anthraquinones in Radix et Rhizoma rhei and their antilipemic effects. MATERIALS AND METHODS: Orthogonal experiment, UV spectrophotometry and conventional antilipemic effect determination method were used to optimize the extraction process, and to determine the total anthraquinone content, as well as blood levels of total cholesterol, triglycerides, HDL and LDL. RESULTS: Ethanol concentration, extraction time and ethanol amount had significant influences on the extraction of total Radix et Rhizoma rhei anthraquinones, total Radix et Rhizoma rhei anthraquinones could significantly reduce blood levels of total cholesterol, triglycerides, HDL and LDL. CONCLUSION: The optimum extraction process was two times extraction of Radix et Rhizoma rhei with 70% ethanol, the amounts of solvent of 8 folds and 5 folds, successively, and the extraction time of 60 min each. In addition, this extract has an antilipemic effect in mice.

Identification and quality evaluation of radix et Rhizoma Rhei by HPLC fingerprint and chemometrics analysis / 中国药学杂志

OBJECTIVE: To develop the HPLC fingerprint of Radix et Rhizoma Rhei and chemometrics analysis method. METHODS: The samples were extracted by ultrasonic with 70% ethanol, purified by 0.45 μm filter membrane, and eluted by 0.1% phosphoric acid and acetonitrile in gradient mode; reference substances and LC-MS were used to identify the chromatography peaks; the HPLC fingerprints of Radix et Rhizoma Rhei and its decoction pieces at 254 and 430 nm were compared and analyzed with CHROMAP software. RESULTS: Authentic and unauthentic Radix et Rhizoma Rhei all have their own characteristic peaks at 254 nm, but the characteristic peaks decreased at 430 nm and the proportion of common peaks areas were obviously different. Twenty-five chromatographic peaks at 254 nm and 19 peaks at 430 nm were selected to distinguish the authentic and unauthentic Radix et Rhizoma Rhei by similarity analysis as well as to evaluate the quality of crude drugs and decoction pieces. CONCLUSION: This method can be used for the identification and quality evaluation of different species of Radix et Rhizoma Rhei crude drugs and decoction pieces.

Review on Research of Compatibility Mechanism of Chemical Ingredients in Prescriptions with Radix et Rhizoma Rhei / 世界科学技术-中医药现代化

Radix et Rhizoma Rhei, the major usage of which is purgation, is widely used in clinical therapy among different prescriptions. According to present reports, researches on herbal compatibility mechanism were mainly fo-cused on the comparison of content changes of active ingredients before and after. In oder to provide references, this review was made on researches of compatibility mechanism of prescriptions and paired drugs for ingredients of Radix et Rhizoma Rhei from research actuality.

Study on Conversion of Conjugated Anthraquinone in Radix et Rhizoma Rhei by Yeast Strain / 世界科学技术-中医药现代化

A yeast strain KM12 , which can convert conjugated anthraquinone of rhub arb to free anthraquinone , was screened. And it is identified as Kluyveromyces marxianus by 26S rDNA. The percentage of free an-thraquinone in total anthraquinone of rhub arb as an indicator was used to investigate the effect of culture medium component and fermentation processing conditions, such as seed age, fermentation time and liquid volume on con-version of conjugated anthraquinone by KM12. The optimal fermentation medium was determined by orthogonal test L9(43) and the composition (%) is yeast extract 1, glucose 2, and rhubarb 2. The 5% (V/V) of the seed culture inoculated 36-48 h was inoculumed to fermentation medium in shake flask to ferment at 30℃, 200 r?min-1 for 4 days. TLC analysis showed most of conjugated anthraquinone decomposed or converted into free anthraquinone in fermented rhub arb . It was concluded that the side effect of severe diarrhea caused by Chinese medicine rhub arb can be alleviated through fermentation processing by KM12 strain .

Bacteriostatic Action of Yiqing Capsule against Propionibacterium Acnes and Staphylococcus Epidermidis in vitro / 中国药房

OBJECTIVE: To study the bacteriostatic action of Yiqing capsule against propionibacterium acnes and staphylococcus epidermidis in vitro.METHODS: Propionibacterium acnes and staphylococcus epidermidis were cultured by microplating method and routine plating,and the bacteriostatic action of different concentration of Yiqing capsule and its main components against propionibacterium acnes and staphylococcus epidermidis were determined by measuring their MICs.RESULTS: The MICs of Yiqing capsule and Scutellaria baicalensis extractum against propionibacterium acnes and staphylococcus epidermidis were 0.98 mg?mL-1 and 1 mg?mL-1,respectively,whereas the MICs of Radix et Rhizoma Rhei and Rhizoma Coptidis Scutellaria extractum against propionibacterium acnes and staphylococcus epidermidis were 1.96 mg?mL-1 and 1 mg?mL-1,respectively.CONCLUSION: Yiqing capsule showed satisfactory bacteriostatic action against the major pathogenic bacteria of acne.

Analysis of Prescriptions on the Compatibility of Chinese Patent Medicines Containing Salvia Miltiorrhiza,Radix et Rhizoma Glycyrrhizae and Radix et Rhizoma Rhei with Western Medicines / 中国药房

OBJECTIVE:To improve the rational compatible use of Chinese patent medicines with western medicines. METHODS:A total of 24 000 prescriptions were randomly sampled from July 10th to July 15th in 2006 for an analysis of the compatibility of Chinese patent medicines containing Salvia miltiorrhiza,Radix et Rhizoma Glycyrrhizae,Radix et Rhizoma Rhei with western medicines.RESULTS:Of the total 24 000 prescriptions analyzed,6 830(or 3 213 patients) involved combined use of Chinese patent medicines with western medicines,of which,the irrational combination for Salvia miltiorrhiza-contained,Radix et Rhizoma Glycyrrhizae-contained and Radix et Rhizoma Rhei-contained Chinese medicines with western medicines totalled 128,118 and 16 cases respectively.CONCLUSION:When the Chinese patent medicines being used in combination with western medicines,their physicochemical and pharmacological properties should be taken into fully consideration so as to achieve rational combination.

Study on Quality Standard of Charred Radix et Rhizoma Rhei / 中国中医药信息杂志

Objective To establish the quality standard of charred Radix et Rhizoma Rhei. Methods Kromasil-C18 column (4.6 mm?150 mm, 5 ?m) was used with mobile phase of methanol-0.1% phosphate acid (85∶15), 1.0 mL/min, and detection at 254 nm. Result Average recoveries of charred Radix et Rhizoma Rhei: Emodin was 98.43% and Chrysophanol was 98.32%, RSD was 1.17% and 1.02% respectively. Conclusion This method is simple. The result is accurate and reproducible. It can be used in the study of the processing technics of Radix et Rhizoma Rhei and the quality control of charred Radix et Rhizoma Rhei.

Determination of Radix et Rhizoma Rhei in Roufuyuliecuotie by HPLC / 中国中医药信息杂志

Objective To establish a method for the determination of Radix et Rhizoma Rhei in Roufuyuliecuotie. Methods The sample was extracted with methyl and the chromatographic condition was as follows :C18 chromatographic column (250 mm?4.6 mm, 5 ?m), a mobile phase of acetonitrile-water- glacial acetic acid (60∶40∶l), the detection wavelength at 437 nm and the flow rate at 1.0 mL/min. Results A linearity of Radix et Rhizoma Rhei in Roufuyuliecuotie was obtained at range of 0.014～0.14 ?g, r =0.9996 (n =6). The average recovery was 98% and RSD=1.47% (n=6). Conclusion This method is easy, sensitive and accurate for the determination of Radix et Rhizoma Rhei in Roufuyuliecuotie.

Influence of Decoction Time on The Total Count of The Total Free Anthraquinone in Radix et Rhizoma Rhei / 浙江中医药大学学报

[Objective]To determine the relation between the total count of the five free anthraquinone(A loe-emodin,Rhein,Emodin,Chrysophanol,Physcion) in Radix et Rhizoma Rhei and the decoction time.[Method]HPLC was used with the chromatographic conditions as follows:mobile phase:methanol-0.1% phosphonic acid solution = 85 :15,detection wave length:254nm,flow rate:1.0 ml/min,column temperature:room temperature.The count of the five free anthraquinone in the decoction taken in every 2 minutes after the solution boiled was determined.[Results]During the 22 minutes after the solution was boiled,the total count took on a ascending tendency,afterwards,it took on a descending tendency.[Conclusion]The biggest number of the total count was obtained 20min after the solution was boiled.

Content Determination of Total Sugar and Polysaccharide in Radix et Rhizoma Rhei and Its Processed Product / 中国药房

OBJECTIVE:To investigate the content of total sugar and polysaccharide in Radix et Rhizoma Rhei and its processed product in order to study the effect of different processing method on the content of total sugar and polysaccharide.METHODS:UV spectrophotometer was adopted to determine absorbance after the sulfuric acid-phenol coloration.The detection wavelength was set at 491 nm.RESULTS:The descending order of the content of total sugar and polysaccharide in Radix et Rhizoma Rhei and its processed product were as follow:prepared Radix et Rhizoma Rhei(steaming),prepared Radix et Rhizoma Rhei(processing with wine),Radix et Rhizoma Rhei,prepared Radix et Rhizoma Rhei with wine,prepared Radix et Rhizoma Rhei with vinegar and charred Radix et Rhizoma Rhei.CONCLUSION:The content of total sugar and polysaccharide in Radix et Rhizoma Rhei processed with different processing method were strikingly different from that of Radix et Rhizoma Rhei.The content of total sugar and polysaccharide in prepared Radix et Rhizoma Rhei were increased significantly while that of prepared Radix et Rhizoma Rhei with wine,prepared Radix et Rhizoma Rhei with vinegar and charred Radix et Rhizoma Rhei were decreased to some extent.

Averse Effect of Radix et Rhizoma Rhei in Ancient and Modern Times / 中华中医药杂志

To know about the toxicity of Radix et Rhizoma Rhei objectively by reviewing the ancient records of toxicity of Radix et Rhizoma Rhei and modern research of its adverse effect.At the same time the reasons of the adverse effect of Radix et Rhizoma Rhei were discussed based on its experiments and clinical practice to provide some reference for its research and reasonable application.

Technology for Optimizing Extraction Method and Uniform Design of Anthraquinones Components of Radix et Rhizoma Rhei / 中国药房

OBJECTIVE: To study extraction method of anthraquinones components of Radix et Rhizoma Rhei and optimize extraction technology.METHODS: Ultrasonic extraction,heating reflux extraction and soxhlet extraction were adopted to extract Radix et Rhizoma Rhei respectively.3 kinds of extraction technology were compared with the content of total anthraquinone,aloe-emodin,rhein,emodin,chrysophanol,physcione as index.The extraction technologies of UV and 5 kinds of anthraquinone compounds were optimized by uniform design method on the basis of previous study.RESULTS: Heating reflux extraction which was optimal extraction technology was as follows: extracting time of 90 min,extracting for 1 time,methanol concentrations of 95%,medicinal mesh of 2.000 7.Comprehensive score of anthraquinones could reach 6.556.CONCLUSION: The extraction technology is reasonable,available in quality control.

Protective Effect of Extracts from Radix et Rhizoma Rhei and Alisma orientalis against Acute Kidney Injury Induced by Diethylene Glycol / 中国药房

OBJECTIVE:To explore the protective effect of the extracts from Radix et Rhizoma Rhei and Alisma orientalis against acute kidney injury induced by diethylene glycol (DEG) and its action mechanism. METHODS: The acute kidney injury model of mice was induced by intragastric administration of DEG. Then the model mice were administered intragastrically with the extracts of Radix et Rhizoma Rhei and Alisma orientalis. Serum levels of creatinine(Cr) and blood urea nitrogen (BUN), the activities of superoxide dismutase (SOD) and glutathione peroxidase(GSH-PX) and the content of malonaldehyde(MDA) were determined. RESULTS: After administration of DEG, the mice presents with obvious toxic reactions including the increase of the ratio of kidney to body weight, the increase of serum levels of BUN and Cr, the decrease of the activities of SOD and GSH-PX in kidney tissue, the increase of the content of MDA. However, after treatment with the extracts of Radix et Rhizoma Rhei and Alisma orientalis, the toxic symptoms in mice were attenuated markedly, the ratio of kidney to body weight decreased, serum levels of BUN and Cr decreased significantly, the activities of SOD and GSH-PX in renal tissues increased and the content of MDA decreased. CONCLUSION: The remarkable protective effect of the extracts from Radix et Rhizoma Rhei and Alisma orientalis against DEG-induced acute kidney injury in mice might be related to its actions of improving the activity of antioxidase of kidney while inhibiting lipid peroxidation.

Study on Preparation and Quality Standard of Flu Mixture / 中国药房

OBJECTIVE: To prepare Flu mixture and establish its quality standard. METHODS: The methods of decoction- boiling and distillation were adopted to prepare the flu mixture; TLC was used to identify Radix Et Rhizoma Rhei and Rhizoma Curcumae Longae, and HPLC was used to determine the content of Artemisinin. RESULTS: The spots characteristic of Radix Et Rhizoma Rhei and Rhizoma Curcumae Longae. were clearly identified with TCL. A good linearity was seen of Artemisinin in the range of 0. 42～ 2. 10? g( r=0. 999 3) . The recovery rate was 99. 84% ( RSD=2. 15% ) . CONCLUSIONS: The preparation is simple in preparation technique and good in stability. The TLC method is highly exclusive. The HPLC method is simple, accurate, and can be used for the quality control of flu mixture.

HPLC Fingerprint Analysis of Medicinal Slices of Radix et Rhizoma Rhei / 中药新药与临床药理

Objective To establish a fingerprint analysis method for the medicinal slices of Radix et Rhizoma Rhei.Methods HPLC with ZORBAX Eclipse XDB-C18 was applied.The chromatographic conditions were as follows:acetonitrile-0.1 %H3PO4 solution(gradient alution) as a mobile phase and detection wavelength at 254 nm,flow rate at 1 mL?min-1,and column temperature being 40 ℃.Results Twenty-five common peaks were separated from 10 batches of medicinal slices of Radix et Rhizoma Rhei.Conclusion The method is reliable and accurate,and can be used as a quality control method for the medicinal material of Radix et Rhizoma Rhei.