Study on material basis of cathepsin K targeted inhibitor in Erzhi Wan / 药学实践杂志

Objective To investigate the active ingredients and components that inhibiting cathepsin K activity in Erzhi Wan, a classic kidney-tonifying formula. Methods Then-butanol, dichloromethane, ethyl acetate and petroleum ether parts and 30 active components in Erzhi Wan were screened by established high throughput fluorescence methods of inhibit the binding activity of CTSK with Z-FR-MCA substrate, the formation of CTSK and chondroitin sulfate A (CSA) complex activity, and the activity of substrate type I collagen degradation by CTSK. Molecular docking and insoluble collagen substrate binding assays were applied to verify the potential CTSK inhibitors. Results The n-butanol and petroleum ether parts of Erzhi Wan inhibited the formation of CTSK and CSA* complex by more than 90%, the petroleum ether part inhibited the binding of CTSK to substrate Z-FR-MCA by more than 90%, the collagen degradation inhibition rate of CTSK in n-butanol part was more than 95% and that in petroleum ether part was 58.6%. Among the 30 active components, 11 showed that the inhibition rate of CTSK and CSA* complex formation was more than 50%, and 5 components with the inhibition rate of Z-FR-MCA binding activity more than 50%. Finally, there were four components including eclalbasaponin Ⅸ, (-)-epicatechin gallate, nuezhenoside and wedelolactone. The inhibition rate of collagen degradation was more than 50%. Eclipta saponin IX inhibited the binding rate between collagen fibers and CTSK, up to 60%, but all of them failed to dock with CTSK active site. Conclusion There are active components that inhibiting cathepsin K in Erzhi Wan, which mainly exists in the n-butanol ingredients, but the active components is not an active-site inhibitor. It might inhibit the binding of CTSK with oligosaccharides by binding to other sites of CTSK, and then reduce the collagen degradation activity of CTSK.

Screening of active fractions of Jiegu ointment for promoting fracture healing in New Zealand rabbits / 中国药房

OBJECTIVE To screen the active site of Jiegu ointment in promoting fracture healing in New Zealand rabbits. METHODS The ethanol extract of Jiegu ointment， as well as the ethyl acetate and n-butanol parts， were prepared and mixed with honey to form a plaster with appropriate viscosity. The radial fracture model of left forelimb in New Zealand rabbit was established and divided into model control group， ethanol extract group， ethyl acetate fraction group and n-butanol fraction group， with 6 rabbits in each group. Except for model control group， rabbits of all other groups were treated with corresponding polar part of Jiegu ointment for external application， for 4 weeks. The radial fracture healing of rabbits was studied by X-ray examination. Enzyme-linked immunosorbent assay was used to detect the serum levels of interleukin-6 （IL-6）， tumor necrosis factor α （TNF-α）， osteocalcin （OC）， vascular endothelial growth factor A （VEGFA）， basic fibroblast growth factor 2 （bFGF2） and alkaline phosphatase （ALP）. HE staining was adopted to observe the changes of pathological morphology of rabbit fracture site， and immunohistochemical method was used to detect the protein expression of bFGF2 in fracture site of rabbits. RESULTS The healing speed of the fracture site in the n-butanol fraction group was the fastest， followed by ethanol extract group， and the ethyl acetate fraction group was the slowest； the serum levels of TNF-α and IL-6 in n-butanol fraction group decreased the fastest， while the levels of ALP， bFGF2， OC and VEGFA increased the fastest [significant increase compared with ethanol extract group （P＜0.01）]； the chondrocytes at the fracture fraction completely disappeared， forming a large number of bone marrow cavities， and the bone trabeculae in the bone marrow cavity were officially formed. The expression level of bFGF2 was also higher than ethanol extract group. CONCLUSIONS The effect of n-butanol fraction on promoting fracture healing is more significant than ethyl acetate fraction and ethanol extract， and n-butanol fraction is the active fraction of Jiegu ointment to promote fracture healing.

Inhibition effect of active fraction from clove on PI3K/Akt/mTOR signaling pathway to induce apoptosis of human colon cancer HCT116 cells / 中国中药杂志

To screen the sensitive cell lines of active fraction from clove(AFC) on human colon cancer cells, investigate the effects of AFC on the cells proliferation and apoptosis as well as PI3 K/Akt/mTOR(phosphoinositide 3-kinase/Akt/mechanistic target of rapamycin) signaling pathways involved, and reveal the mechanism of AFC for inducing apoptosis of human colorectal carcinoma cells. Cell counting kit-8(CCK-8) assay was used to detect the cytotoxic effect of different concentrations of AFC. AFC-induced apoptosis was detected by Hoechst 33258 fluorescence staining and Annexin V-FITC/PI double staining. HCT116 cells were treated with AFC with or without pretreatment with insulin-like growth factor-Ⅰ(IGF-Ⅰ), and then the protein expression levels of caspase-3, caspase-9, poly ADP-ribose polymerase(PARP), PI3 K, p-PI3 K, Akt, p-Akt, mTOR and p-mTOR in PI3 K/Akt/mTOR signaling pathway were detected by Western blot. RESULTS:: showed that the most obvious inhibitory effect of AFC was on human colon cancer HCT116 cells, and the optimal AFC treatment time was 48 hours. After AFC treatment, typical apoptotic features such as nuclear chromatin concentration, nuclear fragmentation and apoptotic bodies appeared in a dose-dependent manner. Annexin V-FITC/PI double staining showed that as compared with the control group, 50 and 100 μg·mL~(-1) AFC groups increased the apoptosis rate of HCT116 cells significantly(P<0.001); AFC activated caspase-9, cleaved caspase-3 and cleaved PARP in a concentration-dependent manner. The protein expression levels of cleaved caspase-3/procaspase-3, cleaved PARP/PARP and caspase-9/β-actin after treatment of AFC(100 μg·mL~(-1)) were significantly different from those in the control group(P<0.001). The relative protein expression of p-PI3 K, p-Akt and p-mTOR decreased in a concentration dependent manner, while Akt and mTOR showed no significant differences among groups. The ratios of p-PI3 K/PI3 K, p-Akt/Akt and p-mTOR/mTOR in the AFC groups(50 and 100 μg·mL~(-1)) were significantly lower than those in the control group(P<0.01). Its combination with IGF-Ⅰ weakened the effect of AFC in inhibiting PI3 K/Akt/mTOR signaling pathway. The ratios of p-Akt/Akt and p-mTOR/mTOR in the AFC+IGF-Ⅰ group were significantly enhanced as compared with the AFC group(P<0.05). Apoptosis-related protein expression levels(cleaved caspase-3 and cleaved PARP) in HCT116 cells treated with AFC+IGF-Ⅰ were also down regulated. As compared with the AFC group, the ratios of cleaved caspase-3/procaspase-3 and cleaved PARP/PARP in the AFC+IGF-Ⅰ group were significantly decreased(P<0.01). In summary, AFC activated caspase-mediated cascades and induced HCT116 cells apoptosis in a dose-dependent manner, which may be associated with the inhibition of the PI3 K/Akt/mTOR signaling pathway.

Screening of Hepatoprotective Activity Fractions from Ixeris chinensis and Study on Its Chemical Compositions / 中国药房

OBJECTIVE： To screen the hepatoprotective active fractions from Ixeris chinensis and study its chemical constituents. METHODS：The petroleum ether，ethyl acetate，n-butanol and residual water fractions from 70％ ethanol extract of I.chinensis were extracted by systematic solvent method. Human hepatocytes HL-7702 were induced by acetaminophen to induce liver injury model. MTT method was used to detect the protective effect of the above fractions（40 μg/mL，by the dosage of crude drug）on injured cells，and the active fractions were screened. The active fractions were separated and purified by silica gel column and Sephadex column chromatography. The structure of the compounds were identified by physical and chemical properties and spectral data （hydrogen spectrum，carbon spectrum）. RESULTS：After treated with different fractions of I. chinensis，the cell survival rate of each administration group was increased significantly，compared with model group（P＜0.01），and the n-butanol and water fractions had the strongest activity （the cell survival rates were 49.3％ and 52.2％ ，respectively）. Six compoundswere isolated from n-butanol fraction and identified as sonchifolignan A（Ⅰ），apigenin-7-O-β-D-glucopyranoside methyl ester（Ⅱ），luteolin-7-O-β-D-glucuronopyranoside methyl ester （Ⅲ），luteolin-7-O-β-D-glucopyranoside （Ⅳ），apigenin-7-O-β-Dglucopyranoside（Ⅴ）and luteolin（Ⅵ）. CONCLUSIONS：The n-butanol fraction is regarded as an effective position for protecting liver，and flavonoids are the main active omponents.KEYWORDS Ixeris chinensis；Hepatoprotective activi

Effect and Mechanism of Active Fraction of Polyrhachis vicina in Treatment of Systemic Lupus Erythematosus(SLE) / 中国实验方剂学杂志

Objective: To observe the effect of an active fraction of Polyrhachis vicina (AFPV) on systemic lupus erythematosus (SLE) and its possible mechanism based on animal and cell models. Method: Totally 60 SD rats were randomly divided into normal control group, model group, prednisone acetate group (5 mg·kg-1), and high, medium and low-dose AFPV groups (400, 200, 100 mg·kg-1). SLE model was replicated with bovine serum albumin-Freund's complete (incomplete) adjuvant. Arthus reaction was observed to study the effect of AFPV on the diameter of back skin redness in rats with SLE. The expressions of anti-double-stranded DNA (dsDNA) antibody, complements 3 (C3), complement 4 (C4), immunoglobulin M (IgM), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-31 (IL-31) and interleukin-33 (IL-33) in serum were detected by enzyme-linked immunosorbent assay. CD4+T cells were isolated from the spleens of MRL/lpr and C57BL/6J mice at the age of 16 to 18 weeks by immunomagnetic beads method. The expressions of miR-200a and miR-155 and the levels of zinc-finger-enhancer binding protein 1(ZEB1) and suppressor of cytokine signaling1(SOCS1) in CD4+T cells were observed to explore the effect of AFPV on SLE and its possible mechanism. Result: Compared with the normal group, the diameter of back skin swelling in the model group was significantly increased (PPPPPPP+T cells of MRL/lpr lupus mice. Compared with the model group, the expression of microRNA-200a increased significantly, the expression of microRNA-155 decreased significantly (PPConclusion: AFPV has therapeutic effect on rats with SLE, its mechanism may be related to the regulation of miR-200a/ZEB1 and miR-155/SOCS1.

Quantitative Determination of Four Active Constituents in Anti-inflammatory and Analgesic Active Fraction of Gaultheria leucocarpa var. yunnanensis / 中国实验方剂学杂志

Objective:To establish a high performance liquid chromatography (HPLC) method for determination of four active constituents, namely MSTG-A,MSTG-B,gaultherin and chlorogenic acid in the anti-inflammatory and analgesic active fraction (ARF) of the ethnic medicine Gaultheria leucocarpa var. yunnanensis, in order to provide a methodological basis for the in-depth study and quality control of G. leucocarpa var. yunnanensis,and lay a foundation for later preparation and clinical application. Method:The determination was performed on COSMOSIL 5C18-PAQ (4.6 mm×250 mm,5 μm) column with methanol-0.2% glacial acetic acid (gradient elution) as the mobile phase at a flow rate of 1 mL·min-1. The column temperature was 25℃. The detection wavelength was set at 294 nm. Result:The linear range of MSTG-B,MSTG-A,gaultherin and chlorogenic acid were 0.014 06-0.450 00,0.007 81-0.250 00,0.003 13-0.100 00,0.000 94-0.030 00 g·L-1 (r ≥ 0.999 7),respectively,with a good precision,repeatability and stability. And the average recoveries were 100.81%,98.99%,96.12% and 102.56%,respectively. RSDs were 1.4%,0.7%,0.7%,2.4%,respectively. The contents of MSTG-B,MSTG-A,gaultherin and chlorogenic acid in ARF fraction of G. leucocarpa var. yunnanensis were 23.608,41.973,8.282,2.673 mg·g-1,respectively. Conclusion:The established method was simple and accurate, with a high repeatability. It can be used for determination of four active constituents in ARF fraction of G. leucocarpa var. yunnanensis,so as to provide a reference for the in-depth research,quality control and comprehensive evaluation of G. leucocarpa var. yunnanensis and lay a solid foundation for preparation and clinical application.

Antitussive effect and mechanism study of effective fractions from Sauropus rostratus / 中草药

Objective: To study the antitussive effect of Sauropus spatulifolius, screen the active part of its antitussive effect, and study its antitussive mechanism. Methods The acute toxicity of different extraction sites of S. spatulifolius were studied by modified Karber’s method; The model was made with ammonia liquor to induce cough. The spray time that caused half of the mice to cough was calculated by sequential method with aim to screen the active sites. Capsaicin was used to induce cough, and the mechanism of action of extracts from various parts of S. spatulifolius on opioid receptor and ATP-sensitive potassium channel (KATP) of mice was explored. Results The LD50 of 75% ethanol, ethyl acetate, n-butanol, and 95% ethanol extracts was 7.30, 17.00, 69.68, and 75.88 g/kg, respectively; The maximum tolerance dose (MTD) of petroleum ether extracts was 117.71 g/kg; Extracts from 75% ethanol and ethyl acetate had antitussive effects, and its antitussive effect was related to opioid receptor and KATP pathway. Conclusion The fractions from 75% ethanol and ethyl acetate are the active parts of S. spatulifolius for relieving cough, and its antitussive mechanism is related to the KATP pathway and opioid receptors in the excited central system.

Identification of Chemical Components in Active Fraction of Xiaozhong Zhitong Lotion by UPLC-Q-TOF-MS / 中国药房

OBJECTIVE： To analyze chemical components in active fraction of Xiaozhong zhitong lotion， to clarify the material basis of its efficacy， and to provide reference for the second development of ointment preparation. METHODS： UPLC-Q-TOF-MS was adopted to analyze the chemical components of active fraction （40％ ethanol elution site separated by D101 macroporous resin） of Xiaozhong zhitong lotion. The determination was performed on Hypersil GOLD aQ C18 column with mobile phase consisted of 0.1％ formic acid acetonitrile （A）-0.1％ formic acid water （B） （gradient elution） at the flow rate of 0.4       mL/min. The sample size was 4 μL， and the column temperature was 30 ℃. The condition of mass spectrometry was ESI detection in positive and negative scanning ion mode （ESI+/ESI－）. The scanning range was 100-2 000 Da. The collision energy was 45/－45 eV， and the energy of the extended collider was 10/15 eV. The accurate molecular weight， retention time and multi-stage fragment ion information of the compounds were collected after obtaining the chromatogram， and the chemical components were identified by comparing with the mass spectrum information of reference materials and references. RESULTS： A total of 48 compounds were identified， and 9 and 39 compounds were identified under ESI+/ESI－ ion mode， mainly including 10 phenolic acids， 8  phenylpropanoids， 9 anthraquinones， 3 flavones， 7 alkaloids， 5 tannins and 6 other categories. CONCLUSIONS： UPLC-Q-TOF- MS method is rapid， efficient and accurate for identify chemical components from active fraction of Xiaozhong zhitong lotion. Main chemical components of the active fraction are phenolic acids， phenylpropanoids， anthraquinones， alkaloids and tannins.

Chemical constituents from active fraction in roots of Euscaphis japonica with inhibitory activity of hepatic triglyceride accumulation / 中草药

Objective: To screen the active fraction of Euscaphis japonica roots with inhibitory activity on hepatic lipid accumulation and investigate its chemical constituents. Methods: Different polar fractions were prepared by extraction with organic solvents, oil Red O staining and triglyceride content assay were used to determine inhibitory activity of different polar fraction on oleic acid induced triglyceride accumulation on HepG2 cells, and the constituents of active fraction were isolated and purified by various chromatography techniques such as column chromatography on silica gel, Sephadex LH-20, and HPLC, and their structures were identified by physicochemical properties and spectral data. Results: The petroleum ether fraction exhibited significantly inhibitory activity on oleic acid induced triglyceride accumulation on HepG2 cells. Nine compounds were isolated and identified as 3β, 19- dihydroxy-24-trans-ferulyloxyurs-12-en-28-oic acid (1), β-sitosterol (2), 7-hydroxy-2-octen-5-olide (3), 3, 3'-dimethoxy-ellagic acid (4), vanillin (5), ethyl-5-oxo-tetrahydro-3-furancarboxylate (6), gallic acid (7), 3, 3'-di-O-methylellagic acid 4-(5″-acetyl)-α-L- arabinofuranoside (8), and bergapten (9). Conclusion: The petroleum ether fraction is main active fraction. The compounds 1, 6, 8, and 9 are obtained from genus Euscaphis Sieb. et Zucc. for the first time.

Study on HPLC fingerprint and quality control of analgesic and anti-inflammatory effective parts in crude and processed roots of Aconitum sinomontanum / 中草药

Objective: To set up an analysis method of fingerprints for analgesic and anti-inflammatory effective parts from crude and processed roots of Aconitum sinomontanum (AS), and to discuss the chemical composition changes after processing that enhance the analgesic and anti-inflammatory effect. Methods: HPLC gradient elution method was developed to establish fingerprints for 10 batches of chloroform extract from crude and processed the roots of AS in different areas. And the fingerprint were analyzed and compared by Chinese Materia Medica (CMM) Fingerprint Similarity Evaluation System (2012 edition). Results: The fingerprints of chloroform extract from crude and processed the roots of AS were set up by HPLC. The gained 3 and 15 common peaks from crude and processed roots of AS, respectively. processed product added 12 peaks, including 1 peak, 2 peak, 7 peak increase were significant, accounting for the new peak area of 72.3%-84.5%. And determination of lappacontine and ranaconitine of chloroform extract from crude and proceed products, after processing the content were reduced, ranaconitine content reduced to the original one-third. Conclusion: This method with good reproducibility, and strong characteristic, and could be used for the full quality evaluation of analgesic and anti-inflammatory effect parts from crude and processed the roots of AS. To provide scientific basis for elucidating the chemical substance base and processing principle of crude and processed roots of AS.

Exploration on extraction and enrichment process of anti-inflammatoryand analgesic active-fraction(ARF)which containsmethyl salicylate glycosides in Dianbaizhu / 国际药学研究杂志

Objective The active ingredient was used as index to optimize the extraction and enrichment process of anti-in-flammatory and analgesic active-fraction(ARF)of Dianbaizhu. Methods Methyl salicylate triglycoside-B was chosen as index com-ponent to extract and enrich methyl salicylate glycosides. Extraction and elution solvents were optimized. The HPLC fingerprint was ob-tained with Thermo Hypersil Gold C18(250 mm×4.6 mm,5μm)column and a gradient elution with the mobile phase consisting of ace-tonitrile(A)-0.2％acetic acid(B)at a flow rate of 1.0 ml/min. And the detection wavelength was set at 294 nm. Results The opti-mized extraction solvent of Dianbaizhu was the 30％ethanol and the optimized elution solvent of ARF enriched by AB-8 macroporous resins was the 35％ethanol. The methodological study on similarity and RSD in ARF HPLC fingerprint of three batches of samples cor-responded to related regulations. Conclusion The extraction and enrichment process of ARF is stable and repeatable.

Antioxidant activity, antibacterial potential and characterization of active fraction of Dioscorea pentaphylla L. tuber extract collected from Similipal Biosphere Reserve, Odisha, India

ABSTRACT Dioscorea pentaphylla L., a wild tuber is used both as food and medicines among different ethnic groups of Similipal Biosphere Reserve, India. Tubers are used against skin infections. In order to establish and confirm tribal claims, methanol extract was subjected to fractionation. The active fraction (DP1) was subsequently used for further purification and NMR (Nuclear magnetic resonance) characterization. The phytochemical analysis revealed the presence of saponin groups. The antibacterial activity of DP1 was done against selected bacterial strains (Salmonella typhi, Shigella flexneri, Streptococcus pyogenes, Streptococcus mutans and Vibrio cholerae) using DD (disc diffusion), AWD (agar well diffusion) and broth dilution assay. The activity was compared with antibiotics Penicillin and Kanamycin. It was observed that DP1 showed significant inhibitory activity against the tested bacteria. The characterization of DP1 through NMR analysis and presence of proton in carbon position at C-3, C-19, C-18, C-21 and C-27 was same as the known compound "Diosgenin". Therefore, isolated compound was confirmed to be Diosgenin. The study for the first time showed that, diosgenin present in D. pentaphylla tuber was responsible for antibacterial and antioxidant potential. Present study highlights the importance of Dioscorea species as sources of diverse secondary metabolites for the isolation of active compound(s).

Identification of a fat-soluble fraction of Sparganium stoloniferum that inhibits cervical cancer HeLa cells

ABSTRACT Context The rizoma of Sparganium stoloniferum has been used as a traditional Chinese medicinal herb for thousands of years. Sparganium stoloniferum is a stasis-breaking drug to treat a wide range of diseases including cancer,however, its activity of extract on cervical cancer HeLa cells and the mechanisms remains unknown. Objective This study aimed to screen Sparganium stoloniferum extract for its inhibitory effects on cervical cancer HeLa cells. Materials and methods Sparganium stoloniferum was extracted with 95% ethanol under reflux, and the extracts were preliminary separated by silica gel column chromatography. MTT assay and flow cytometry were used to determine the inhibitory effects of three fractions on HeLa cells. In addition, GC-MS was performed to analyze the chemical composition of the active fraction. Results Sample II showed a dose-dependent inhibition of HeLa cell growth, with an inhibition rate of more than 30%, whereas the inhibition rates of Samples I and III were less than 30%. Interestingly, Samples I and III had no effect on apoptosis, in contrast to Sample II, which significantly promoted HeLa cell apoptosis, in a dose-dependent manner. GC-MS was performed to analyze the chemical components of Sample II: steroids were found to be the major components with a relative content of 73.905%, while six known compounds were obtained for the first time. Discussion and conclusion This study provided a novel insight for further research of active fractions of Sparganium stoloniferum, in accordance with the basic principle and theory of traditional Chinese medicine (TCM), and will promote the shift of effective substance study, from monomeric chemical compounds to active fractions.

Effects of different penetration enhancers on pharmacokinetics of active components in Xiangfu Siwu transdermal patches / 中国中药杂志

To study the effect of different penetration enhancers on the pharmacokinetic characters of six active components in Xiangfu Siwu transdermal patch (XBW) and optimize the best penetration enhancers. During the experiment, the patches containing different penetration enhancers were stuck on the rat's skin, and then the blood samples were acquired at different time points. Six active components in plasma were determined by UPLC-MS/MS. The main pharmacokinetic parameters were calculated with DAS software package. The total factor scores (F) of the plasma concentrations of six components at every time point in different groups were calculated using principle component analysis, and the areas under F versus time curves (AUCF-t) were employed to be the indexes for selecting penetration enhancers. The results demonstrated that compared with the control group, the AUCF-t from other groups increased prominently and furthermore, 5% menthol manifested the best effect. In this research, 5% menthol could remarkably promote the percutaneous penetration effect of the six active compounds in XBW, and it could provide a scientific basis for the preparation research of XBW.

Anti-inflammatory active fraction screening and mechanism of unripe Forsythiae Fructus based on ¹H-NMR metabolomics / 中国中药杂志

To screen the anti-inflammatory active fraction of unripe Forsythiae Fructus, and elucidate the action mechanism, water decoction, ethyl acetate portion, n-butanol portion and residue water extracts of unripe Forsythiae Fructus were administered into rats for continuously 15 days. The acute lung injury inflammatory model was established to observe the section structure of lung tissues. Levels of IL-6, TNF-α, IL-1β and IL-10 in bronchoalveolar lavage fluid were determined by ELISA kits, and changes in endogenous metabolites in serum were analyzed based on 1H-NMR metabolomics. The results showed that ethyl acetate portion of unripe Forsythiae Fructus had a better anti-inflammatory activity against acute lung injury, and could suppress the release of inflammatory factors of IL-6, TNF-α, IL-1β, significantly reduce contents of creatine, β-OH-butyrate, succinate, lysine, valine, isoleucine and glutamine, and elevate the content of GPC in serum. Ethyl acetate portion was proved to be the main fraction of anti-inflammatory activity from the perspective of endogenous metabolites in serum, and played an anti-inflammatory role by regulating creatine metabolism, choline metabolism, branched-chain amino acid metabolism and TCA cycles. This study could lay a foundation for studying pharmacodynamic material basis of unripe Forsythiae Fructus.

Vasodilation Effect and Underlying Mechanism of Active Fraction from Compound Prunella Vulgaris L. in I-solated Thoracic Aorta of Rats / 中国药师

Objective:To investigate the vasorelaxant effect of active fraction from compound Prunella vulgaris L. ( AFCP) on the i-solated thoracic aorta of rats and underlying mechanism. Methods:The study was performed with the tension experiment of the isolate rat thoracic aorta. The changes of vascular ring tension were measured by biological signal acquisition and analysis system, and the vasodila-tor effect of AFCP was observed. Results:AFCP(100-500μg·ml-1)could induce significant relaxation in aorta rings pre-contracted by phenylephrine (PE,1 μmol·L-1), and the relaxation effect was significant(75% ±8%) in endothelium-intact aortic and endothelium-denuded aortic. The vasodilatation effect of AFCP was not significantly affected by nitric oxide synthase ( NOS) inhibitor NG-nitro-L-argi-nine( L-NNA) , guanylate cyclase inhibitor MB,potassium channel blocker TEA and glibenclamide. In Ca2+-free bath solutions, AFCP (300 μg·ml-1 ) could shift downward dose-response curve of CaCl2 and significantly reduce the maximum contraction amplitude of PE. Conclusion:AFCP can relax rat aorta rings in a dose-dependent manner, which is endothelium-independent. The mechanism may be re-lated to the inhibition of intracellular calcium release and extracellular calcium flow, and has nothing to do with NO pathway, prostacyclin generation and calcium-activated potassium channels.

Study on in vitro transdermal absorption properties of active fraction from Xiangfu Siwu Decoction / 中草药

Objective: To study the in vitro transdermal absorption properties of active fraction from Xiangfu Siwu Decoction (XSD). Methods: UPLC-MS/MS method was established for the determination of six main active ingredients (ferulic acid, paeoniflorin, albiflorin, tetrahydrocolumbamine, protopine, and tetrahydropalmatine) in active fraction of XSD (BW) and transdermal receiving liquid. The active ingredient group of six active ingredients was prepared according to their proportion in BW. The experiment of in vitro transdermal absorption was performed using vitro-abdominal skin of rats and improved Franz diffusion cell method. Results: UPLC-MS/MS method showed perfect specificity and the six ingredients performed well in linear relationship (r>0.997), precision (RSD≤2.66%), repeatability (RSD≤2.98%), stability (RSD≤3.99%), and average recovery rate [(95.22±5.48)%-(103.68±2.90)%]. The cumulative transmittance of the six ingredients displayed the same order between the active ingredient group and BW. The cumulative transmittance in descending order was as follows: ferulic acid>tetrahydrocolumbamine > tetrahydropalmatine>protopine>paeoniflorin≈albiflorin. Additional, transdermal absorption properties of the six ingredients in vitro were affected by the pH value of supply liquid significantly. Conclusion: The in vitro transdermal absorption behavior of the active ingredient group is consistent with BW and pH value can change the behavior of active ingredients in in vitro transdermal absorption.

Effects of Active Fraction of Angelica Sinensis Radix on Immunological Function in Mice under High Altitude Hypoxia Condition / 中国中医药信息杂志

Objective To explore the intervention effect of active fraction of Angelica Sinensis Radix in mice under high altitude hypoxia condition. Methods Totally 72 healthy SPF mice were randomly divided into control group (K), model group (M), Rhodiola rosea group, and active fraction of Angelica Sinensis Radix groups (B, C, X). The mice were administerted corresponding treatment by gavage for 21 days. Control mice were given normal saline in same volume. From the 8th day, all mice excepted control mice were exposed to high altitude hypoxia cabin after 0.5 hour gavage treament. On the 22nd day, after got out of the cabin and their body weight were measured, mice were put to death through eyeball blood sampling to prepare splenic lymphocyte suspension. The proliferation and transformation capacities of lymphocyte cell and killing activity of NK cells were detected by MTT. The content of IL-2 in the serum of mice in each group were detected by ELISA. Results Compared with the control group, the body weight of mice, the proliferation and transformation capacities of lymphocyte cell, the killing activity of NK cells, and the content of IL-2 were all significantly decreased (P<0.05, P<0.01). Experiment tests showed that the proliferation and transformation abilities of lymphocyte cell and the killing activity of NK cells were all increased in the mice of group B, C, and X compared with those of the model group (P<0.05, P<0.01). The stimulate index of lymphocyte cell was raised after X intervention compared with those of the model group (P<0.05). The content of IL-2 in the serum was enhanced after intervention of active fraction C and X of Angelica Sinensis Radix compared with those of the model group (P<0.05, P<0.01). Conclusion Active fraction of Angelica Sinensis Radix shows increasing immunological function of mice exposed to hypoxia.

Phytochemical screening and antihyperglycemic activity of Heliotropium indicum whole plant in Streptozotocin induced diabetic rats.

Present study was designed to screen phytochemical constituents and antihyperglycemic activity of Heliotropium indicum (HI) in Streptozotocin (STZ) induced diabetic rats. Heliotropium indicum (Boraginaceae) whole plant is used as traditional medicine for a number of ailments including diabetes. The whole plant was collected, shade dried and extracted with different solvents in the increasing order of polarity. When different solvent extracts of HI each at a dose of 500 mg/kg bw were given to diabetic rats, the methanol and aqueous extracts produced significant (P<0.0001) antidiabetic activity. Phytochemical screening of various solvent extracts of HI whole plant revealed the presence of alkaloids, steroids, triterpenes, saponins and tannins. When methanol active fraction of Heliotropium indicum (MAFHI) was checked for its antidiabetic activity, the fraction at dose of 750 mg/kg bw produced marked antihyperglycemic activity. The antihyperglycemic activity was also exhibited during oral glucose tolerance test (OGTT) with the same dosage of MAFHI.

Effect of active fraction from Huanglian Jiedu Decoction on activation of astrocytes and expression of Cx43 in cerebral ischemic penumbra / 中草药

Objective: To observe the effect of water extract and active fractions from Huanglian Jiedu Decoction (HJD) (total alkaloids, total favonoids, and total iridoids) on the activation of astrocytes and expression of connexin 43 (Cx43) in ischemic penumbra of rats after focal cerebral ischemia. Methods: The rat model of middle cerebral artery ischemia was established using suture-occluded method. The male SD rats were randomly divided into Sham-operation, model, water extract of HJD (800 mg/kg), total alkaloids (44 mg/kg), total flavonoids (50 mg/kg), and total iridoids (80 mg/kg) groups. They were ig administered for continuous 7 d, once daily. The histopathologic changes of brain in rats were observed by HE staining. The expression of GFAP and Cx43 was detected by immunofluorescence assay and RT-PCR was used to detect the gene expression of Cx43. Results: In the model group, the number of neurons in ischemic penumbra of rats was significantly decreased and the expression of GFAP, Cx43, and Cx43 protein was obviously increased compared with Sham-operation group (P<0.01). The three active fractions from HJD (total alkaloids, total favonoids, and total iridoids) could increase the number of neurons in the ischemic penumbra of rats and decrease the expression of GFAP, Cx43, and Cx43 protein obviously compared with the model group (P<0.05, 0.01). Conclusion: The active fractions (total alkaloids, total favonoids, and total iridoids) could inhibit the over activation of astrocytes and the expression of Cx43, so as to protect the neurons in ischemic penumbra.