The mechanism of effective fractions from Xiongma decoction in the treatment of migraine model rats with hyperactivity of liver-yang and blood stasis / 中国药房

OBJECTIVE To expl ore the mechanism of effe ctive fractions from Xiongma decoction in the treatment of migraine with hyperactivity of liver-yang and blood stasis. METHODS Totally 70 male SD rats were randomly divided into normal group ， model group ，positive control group （Flunarizine hydrochloride capsules 0.9 mg/kg），low-dose and high-dose groups of Xiongma decoction effective fractions （ethyl acetate extract 0.87，3.46 g/kg，n-butanol extract 1.80，7.20 g/kg）. Except for normal group ， rats in other groups were given aconite decoction （2 g/kg），once a day ，for 4 consecutive weeks to establish the hyperactivity model of liver-yang. On the 15th day of modeling ，all administration groups were given corresponding drugs intragastrically at the same time ，once a day ，for 2 consecutive weeks. On the 29th day of modeling ，rats trigeminal ganglion was stimulated to establish the migraine model with hyperactivity of liver-yang and blood stasis ，then the medication was maintained for another time according to the above method. The macroscopic signs and behavior of the rats were observed ；positive expression ，mRNA and protein expression of transient receptor potential vanilloid 1 （TRPV1），calcitonin generelated peptide （CGRP），calcitonin receptor-like receptor （CRLR） and receptor-associated membrane proteins 1 （RAMP1） in trigeminal cervical spinal complex （TCC） were detected by immunohistochemistry ，RT-qPCR and Western blot assay. RESULTS Rats in model group showed macrophysical signs and behaviora l manifestations related to migraine with hyperactivity of liver-yang and blood stasis. Thirty minutes after last administration ，the above conditions of rats in Xiongma decoction effective fraction groups were improved significantly. Compared with normal group ， positive expression，mRNA and protein expression of TRPV 1，CGRP， CRLR and RAMP 1 in TCC of rats in the model group were significantly increased （P＜0.05）. Compared with model zengguirong@hnse.org group， most of above indicators in Xiongma decoction effective fraction groups were significantly decreased （P＜0.05）. CONCLUSIONS The mechanism of Xiongma decoction in preventing and treating migraine with hyperactivity of liver-yang and blood stasis may be related to inhibit the activity of TRPV1-CGRP/CGRP receptor signaling pathway in TCC.

Evaluating the quality of Miao medicine Periploca forrestii Schltr based on 6 caffeoylquinic acid components combined with stoichiometry / 中草药

Objective: To investigate the effects of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C in effective parts of rhizoma arthritis against rhizoma arthritis from different sources of Miao medicine Periploca forrestii Schltr. The determination of 6 kinds of caffeoacylquinic acid chemical components and comprehensive analysis of their quality combined with chemometrics provide a basis for a reasonable and effective evaluation of the efficacy of Miao Medicine Periploca forrestii Schltr against rheumatoid arthritis. Methods: Simultaneous determination was performed by HPLC-UV method. The column was Xtimate C18 (250 mm×4.6 mm, 5 μm), the mobile phase was 0.1% formic acid-acetonitrile, gradient elution, the injection volume was 10 μL, and the flow rate was 1 mL/min. The column temperature is 25 ℃ and the detection wavelength is 327 nm. SPSS24.0 and SIMCA14.1 software are used for comprehensive analysis and evaluation of the quality of medicinal materials. Results: The content of 21 batches of Periploca forrestii Schltr samples from different sources was determined. Through the cluster analysis, the 21 batches of samples were classified into type III, of which the quality of types II and III was better; principal component analysis and partial least squares discriminant analysis The results were consistent with the clustering results. The chlorogenic acid and isochlorogenic acid C were screened to be the markers that caused the differences in the quality of Periploca forrestii Schltr from different sources. Conclusion The determination of the contents of six caffeoacylquinic acid components combined with stoichiometry can further evaluate the efficacy and quality of the anti-rheumatoid arthritis effect of Periploca forrestii Schltr, and provide a reference basis for the quality control of Periploca forrestii Schltr.

Effects and active substances of ethanol extract from Dendrobium officinale on metabolic hypertensive rats induced by comprehensive dietary / 中国中药杂志

Previous studies of Dendrobium officinale on anti-hypertension effect always focused only on the blood pressure,while polysaccharides of D. officinale( DOP) have been traditionally considered as one of the main effective substances. This study aimed to evaluate the effect of ethanol extract from D. officinale( DOE) on blood pressure,Glu and lipid profile in metabolic hypertensive rats induced by comprehensive dietary factors,and elucidate the composition of effective fractions from DOE. A metabolic hypertension model of rat induced by high-sugar,high-fat diet and alcohol drinking was adopted to evaluate the effect of DOE on hypertension and other metabolic disorders. Blood pressure,Glu and lipid profile were detected to find the features and differences of DOE and DOP on metabolic hypertension. Furthermore,DOE was separated with three different common solvents according to the polarity. Along with blood pressure,Glu,UA and lipid profile,hemorheology,oxidative index and aortas structure changes were adopted to evaluate the comprehensive effects of the most effective fractions on metabolic hypertension. Finally,HPLC-DAD-MS was adopted to identify the components of the most effective fraction. The SBP and Glu of models were decreased significantly after administration of DOE and DOP for 6 weeks,while TG in DOE groups also reduced dramatically. The DOE was separated with ether,n-butanol respectively and named NAF,NBF and NCF. SBP,TG,Glu,UA of model rats were decreased significantly after 4 weeks administration with NBF. The level of MDA in serum was down-regulated,while GSH-Px and T-AOC were up-regulated obviously after 12 weeks.And the blood viscosity also obviously decreased,with less collagen deposition of aortas by Masson's trichrome staining. NBF was mainly composed of phenols and flavone C-glycosides,whose aglycone was apigenin,and monosaccharide was connected to C-6 and C-8. Ethanol extract from D.officinale has an positive effect in alleviating hypertension and metabolic disorders in metabolic hypertension. Medium polarity fraction was the effective fraction of alcohol extraction from D. officinale,and mainly composed of phenols and flavone C-glycosides.

Diuretic effect and material basis of Clematidis Armandii Caulis in rats / 中国中药杂志

To search for the active diuretic fractions of Clematidis Armandii Caulis( CAC) and determine its main active chemical components by using liquid chromatography-mass spectrometry( LC-MS) and diuretic activity evaluation. CAC 75% ethanol extracts and extracts from different polar solvents were orally administered to saline-loaded rats at different doses. 6 h urinary volume,p H and contents of electrolyte Na+,K+and Cl-were measured. The chemical components of the active fractions were separated and identified by ultra performance liquid chromatography-electrospray ionization-quadrupole time of flight-mass spectrometry( UPLC-ESI-Q-TOF-MS/MS) method. As compared with the control group,the urine volume was increased by 44%( P< 0. 01) and 34%( P < 0. 05) in CAC75% ethanol extract 57. 74 and 28. 8 mg·kg-1 groups respectively; the Na+excretion was increased by 52%( P< 0. 01) and 45%( P<0. 05),respectively; while the Cl-excretion was increased by 101%( P<0. 01) and 85%( P<0. 05),respectively. The urine volume,Na+excretion and Cl-excretion were increased by 50%( P< 0. 01),58%( P< 0. 05),and 65%( P< 0. 05) respectively in petroleum ether extract 70. 98 mg·kg-1 group as compared with the control group. While for the n-butanol extract 194. 18 mg·kg-1 group,the urine volume,Na+and Cl-excretion were increased by 42%( P<0. 01),41%( P<0. 05) and 97%( P<0. 01),respectively. The diuretic activity of other fractions was not obvious. There was no statistical difference in K+excretion in all groups. The results of LC-MS analysis showed that six compounds,including two sterols,one chromogen and three fatty acids,were identified from petroleum ether extract.Fourteen compounds,including six triterpenoid saponins,six lignin glycosides,one sterol glycoside and one phenolic glycoside,were identified from the n-butanol extract. All the results suggested that the ethanol extract of CAC had remarkable diuretic activity and its main effective components included sterol,triterpenoid saponin and lignin glycosides.

Determination of 13 active components in Huanglian Jiedu Decoction by HPLC and screening of its effective fraction / 中草药

Objective: To establish a method for the determination of 13 active components (phellodendrine, chlorogenic acid, magnoflorine, geniposide, coptisine, epiberberine, jatrorrhizine, berberine, palmatine, baicalin, wogonoside, wogonin, and oroxylin A) in Huanglian Jiedu Decoction (HJD) by HPLC, and screen the effective fractions of HJD by the content and ratio of active components. Methods: The freeze-dried powder of HJD water decoction was extracted by petroleum ether, ethyl acetate, n-butanol and pure water by soxhlet extractor combined system solvent extraction method. The different polar fractions of HJD were obtained after drying, and the residue was used as the precipitation fraction. Subsequently, 13 active ingredients in HJD were detected by Agilent 1260 high performance liquid chromatography. Furthermore, the content of active components in HJD and its different polar fractions were detected, and the effective fraction of HJD were screened. Results: A stable and reliable HPLC method was successfully established, and the content of active components in HJD from high to low were: geniposide, berberine, palmatine, baicalin, wogonoside, coptisine, jatrorrhizine, epiberberine, magnoflorine, phellodendrine, wogonin, chlorogenic acid, and oroxylin A. Compared with the fractions of petroleum ether, ethyl acetate, purified water, and precipitation, the total content of active components in the fraction of n-butanol of HJD was the highest and its proportion is the closest to that of HJD, which was the effective fraction of HJD. Conclusion: In this study, a method for the determination of 13 active ingredients in HJD by HPLC was successfully established, and the n-butanol fraction of HJD was selected as the effective fraction of HJD based on the content of active components.

Screening the Effective Hemostasis Fraction of Balanophora Involucrata / 中国药师

Objective: To screen the effective hemostasis fraction of Balanophora involucrata. Methods: Balanophora involucrata was extracted by 95% ethanol using reflux extraction, and the residue was extracted by water. The filtrate was combined and concen-trated, and then extracted by petroleum ether, ethylacetate and n-butylalcohol in turn to respectively obtain petroleum ether phase, eth-ylacetate phase, n-butylalcohol phase and H2O phase. The effective hemostasis fraction was determined by measuring the time of blood coagulation and hemorrhage combined with plasma recalcification time. Results: Compared with the blank group, ethylacetate extract from Balanophora involucrata could significantly shorten the time of blood coagulation and hemorrhage, and the plasma recalcification time (P<0. 01). n-Butylalcohol group at high dose could shorten the time of hemorrhage and the plasma recalcification time (P<0.05),while had no effect on the time of blood coagulation(P>0.05). Petroleum ether extract and H2O phase couldn't shorten the time of blood coagulation and hemorrhage, and the plasma recalcification time(P>0. 05). Conclusion: The effective hemostasis frac-tion of Balanophora involucrata mainly concentrates in ethylacetate extract.

Effect of Effective Fraction from Radix Vitis Davidii Foex. on Fracture Healing / 医药导报

Objective To investigate the effect of effective fractions from Radix Vitis Davidii Foex. on fracture healing. Methods SD rats were randomly divided into model control group,Yunnan Baiyao group,and effective fraction group (n= 36 each group). Fracture models were established. Rats in the Yunnan Baiyao group and effective fraction group were given 3.0 g?kg-1?d-1 of Yunnan Baiyao and effective fraction from Radix Vitis Davidii Foex.,respectively,and rats in the model control group was treated with the same volume of pure water. Alkaline phosphatase and Ca concentration in serum were determined.For radius fracture structure,digital radiography and RT-PCR detection of VEGF mRNA expression were performed. Results The peak concentration of the alkaline phosphatase and Ca concentration in serum were significantly higher in the effective fraction group than in the Yunnan Baiyao group,and peak value appeared earlier,fracture healing score was higher, fracture line disappeared earlier,and VEGF mRNA expression was significantly higher in three weeks after the fracture (all P<0. 01) . Conclusion The effect of the effective fraction from RadixVitis Davidii Foex.on the fracture healing is strong.

Research Approaches on Discovery R&D of Innovative Drug of Chinese Materia Medica under New Situations / 世界科学技术-中医药现代化

In recent years,with the rapid development of life sciences,changes on the understanding and requirements for innovative drugs of Chinese materia medica (CMM) from the drug approval authorities have been made.In this paper,the discovery,research and development (R & D) approaches for innovative drugs of CMM under new situations were summarized and introduced in details from five aspects as follows.The discovery of innovative drugs from active components of CMMs,active fractions of CMMs and their compound preparations,famous classic prescription,clinically effective prescriptions as well as the famous Chinese patent drugs.This paper puts forward some new ideas for the pharmaceutical researches of innovative drugs of CMM,including the production process,chemical components,quality study,quality standard and stability.This paper provided references for the discovery,R & D of CMM.

Analysis of components in plasma from effective fractions of Inula cappa based on UHPLC/Q-TOF/MS / 中国药理学通报

Aim To analyze the main components of effective fractions of Inula cappa in rat plasma with UHPLC/Q-TOF/MS technology and serum pharmacochemistry theory.Methods After gavage administration with Inula cappa extracts,blood was collected from abdominal aorta,and the protein in plasma sample was precipitated by methanol.Eclipse Plus C18 RRHD (2.1 mm × 100 mm,1.8 μm) was used,with 0.1％ formic acid agueous solution (A)-0.1％ formic acid and acetonitrile (B) as the mobile phase for gradient elution,and the flow rate was 0.3 mL ·min-1 Detected at negative ion mode,and with the help of Metabolite Tools software from Bruker Corporation,components in plasma were defined.Results A total of 16 compounds were identified,including 9 prototypes and 7 metabolites.Main metabolites were isomerization,methylation,glucuronidation and recombination of caffeoylquinic acid.Conclusions Therefore,our study has comprehensively expounded Inula cappa extracts' constituents migrating to rat plasma,and provided a reference for further studies for in vivo metabolic process and effective material base of Inula cappa.

Chemical constituents of effective part in Celosia cristata for treatment of hemostatic / 中草药

Objective: To study the chemical constituents of effective fraction in Celosia cristata for hemostatic. Methods: The chemical constituents were isolated and purified by chromatography on silica gel, ODS, and preparative HPLC methods. Their structures were determined by physicochemical properties and spectral data. Results: Nine compounds have been found in the ethyl acetate fraction of 95% ethanol extraction from C. cristata which were obtained and identified as northalifoline (1), 1,2,3,4-tetrahydro-4-hydroxy-4-quinolin carboxylic acid (2), isorhamnetin-3-O-β-D-glucoside (3), 1-feruloyl-β-D-glucopyranoside (4), Feruloyl 6-O-β-D-glucopyranose ester [5 (α structure), 6 (β structure)], 2-methoxy-4-(2-propenyl) phenyl β-D-glucopyranoside (7), lupeol (8), and ferulic acid (9). Conclusion: All compounds are not only isolated from C. cristata for the first time but also from the plants of genus Celosia L. for the first time.

Separation and Enrichment of Antibacterial Effective Fraction of Syringae Folium by Macroporous Resins / 中国药师

Objective:To investigate the purification of antibacterial effective fraction of Syringae folium by macroporous resins. Methods:Static adsorption and desorption tests were carried out to screen the macroporous resins. The desorption experiment was per-formed on the selected D101 resin to optimize the separation process. The effects of resin amount, diameter length ratio, elution flow rate, elution solution concentration and volume were studied. Results:The optimal conditions were as follows:the elution solution was 55% ethanol, the adsorption flow rate was 1 BV·h-1 , the elution flow rate was 5 BV·h-1 , 6 BV 25% ethanol was used to eliminate impurity and 8 BV 55% ethanol was used to elute to obtain the effective fraction. Conclusion: The content of antibacterial effective component is above 65% after purified by D101 resin, indicating that the present method is suitable for large-scale preparation of anti-bacterial effective fraction of Syringae folium.

Optimization of Extraction Process for Effective Fraction of Bushen Yizhi Formula / 广州中医药大学学报

Objective To optimize the extraction process for the effective fraction of Bushen Yizhi ( BSYZ) formula. Methods Yield of the effective fraction and contents of two marker compounds 2,3,5,4 -tetrahydroxystilbene-2-O-β-D-glucoside and osthole assayed by high performance liquid chromatography (HPLC) were used as the optimizing indexes, and weighted coefficient of the indexes was analyzed by analytic hierarchy process ( AHP). Ratio of liquid-to-solid, extraction time, and extraction times were used to screen the optimal conditions by orthogonal test. Results The optimized extraction process of the effective fraction of BSYZ formula was as follows: extracting with 8-fold water for three times, and boiling for 1.5 hours each time. The verification test showed that the comprehensive scores of the optimized extractive process were higher than those obtained in the orthogonal test. Conclusion The optimized extraction process and conditions are practical in the manufacture of the effective fraction of BSYZ formula.

Protection and mechanism of effective fraction from Buyang Huanwu Decoction on focal cerebral ischemia-reperfusion injury in rats / 中草药

Objective: To study the protection and mechanisms of effective fraction from Buyang Huanwu Decoction (EFBHD) on rat brain after cerebral ischemia-reperfusion injury. Methods: Rat model of cerebral ischemia-reperfusion injury was created by the middle cerebral artery occlusion (MCAO) by modified suture method. Healthy male SD rats were randomly divided into six groups: Sham-operated group, model group, high-, middle-, and low-dose (200, 100, and 50 mg/kg) of EFBHD groups, and positive control group (treated with EGB 100 mg/kg). The neurological deficit symptom scores were observed and the infarct volume was measured by triphenyltetrazolium chloride (TTC) staining in 24 h after the cerebral ischemia. The contents of TNF-α, IL-1β, and IL-6 in serum were measured by enzyme-linked immunosorbent assay (ELISA) methods. In addition, the activity of LDH and MDA content in serum were determined by abdominal arterial blood and centrifuged. Results: Compared with the model group, EFBHD could significantly improve the neruological dysfuction, decrease the cerebral infarct volume, and inhibit the activity of LDH, and reduce the contents of MDA, TNF-α, IL-1β, and IL-6 in serum (P<0.01, 0.05). Conclusion: EFBHD has significant protection on cerebral ischemia-reperfusion injury in rats, which may be related to the inhibition of inflammatory cytokine secretion and expression and inducing the inflammation in brain tissue.

Modernization of Chinese materia medica processing pieces / 中草药

In the modernization course of Chinese materia medica(CMM),two misunderstanding mistakes which are out of line with traditional Chinese medicine(TCM)theory and excessively emphasize the research of CMM compound prescription and two difficult points which are the establishment of CMM quality control system and the establishment of CMM evaluation system were pointed out.Because the main means of TCM syndrome differentiation treatment and TCM combination therapy is CMM processing pieces and the starting point of the research of CMM efficacy substantial basis should be CMM processing pieces,the priority of the modernization of CMM should be the research on modern CMM processing pieces and the conditions have been mature now.The research on modern CMM pieces were supposed to take three steps:the research on "standardized extract",the research on "effective fraction" and the research on the CMM nature of "effective fraction".

Antioxidation and Immuno-regulation Effects of Jiawei Zhisou Powder on Hypersensitive Asthma Mice / 中药新药与临床药理

Objective To study the antioxidation and immuno-regulation effects of the active fraction in Jiawei Zhisou Powder.Methods Hypersensitive asthma mice were established by the miscible liquids of ovalbumin(OA)and Al(OH)3.The influences of the powder on the contents of SOD and MDA in the serum and lung tissue of mice were observed and the contents of IL-4 and IFN-?in supernatant of mouse spleen cells cultured in vitro were measured by enzyme-linked immunosorbent assay.And Jiawei Zhisou Powder and dexamethasone were administered to normal mice respectively,then the immune organs weight were measured and the spleen and thymus indexes of mice were calculated.Results Compared with the normal group,the contents of SOD and IFN-?were significantly decreased,while MDA and IL-4 contents were obviously enhanced in asthma group(P