Research advance on structure and function of amides in Zanthoxylum plants / 中国中药杂志

Zanthoxylum belongs to the Rutaceae family, and there are 81 Zanthoxylum species and 36 varieties in China. Most of the Zanthoxylum plants are used as culinary spice. In recent years, scholars in China and abroad have carried out in-depth research on Zanthoxylum plants, and found that the peculiar numbing sensation of Zanthoxylum plants originates from amides. It is also determined that amides are an important material basis for exerting pharmacological effects, especially in anti-inflammatory analgesia, anesthesia and other aspects. In this paper, 123 amides in 26 Zanthoxylum plants and their pharmacological activity that have been reported were summarized, which provided scientific reference for the clinical application of Zanthoxylum plants and the research and development of new drugs, and also facilitated the sustainable development and utilization of Zanthoxylum plant resources.

Genetic diversity and antibacterial activity of endophytic fungi from Zanthoxylum nitidum / 中国中药杂志

In order to reveal the distribution and population characteristics of endophytic fungi from Zanthoxylum nitidum and the antibacterial potential,this study performed molecular identification and analyzed the genetic diversity and antibacterial activity of endophytic fungi from Z. nitidum in Guangxi. Through culture and molecular identification,35 strains,belonging to 15 genera,12 families,10 orders,4 classes,and 2 phyla,were isolated from various tissues of Z. nitidum,of which Colletotrichum and Fusarium were the dominant genera,respectively accounting for 20% of total strains. The diversity of endophytic fungi was significantly different among roots,stems,and leaves,as manifested by the significantly higher Shannon index( H') in stems( 1. 678) than in roots( 0. 882 1) and leaves( 0. 515 4). The antimicrobial activity analysis showed that 14. 28% of endophytic fungi inhibited at least one indicator pathogen. Among them,Fusarium sp. ZN-34 and Fusarium sp. ZN-26 separately demonstrated the strongest inhibitory effect on Escherichia coli and Staphylococcus aureus. In general,Fusarium sp. ZN-26 and Phialemoniopsis plurioloculosa ZN-35 were advantageous in suppressing the two bacteria owing to the broad spectrum and strong efficacy. In summary,Z. nitidum in Guangxi boasts rich endophytic fungi with the majority showing strong antibacterial activity,which can be used as candidates for the extraction and separation of basic antibacterial substances and the development of natural antibacterial agents.

Extract from Zanthoxylum piperitum Induces Apoptosis of AGS Gastric Cancer Cells Through Akt/MDM2/p53 Signaling Pathway / 中国结合医学杂志

OBJECTIVE@#To determine the effect of Zanthoxylum piperitum extracet (ZPE) on apoptosis and analyze anticancer substances in ZPE, changes in proteins related to apoptosis, and pathological changes in tumors in mouse.@*METHODS@#Fifteen 4-week-old female BALB/c nu/nu mice were divided into 3 groups depending on ZPE dose, with 5 in each group. AGS gastric carcinoma cells (1 × 10@*RESULTS@#High performance liquid chromatography (HPLC) analysis showed that ZPE contained organic sulfur compounds such as alliin and S-allylcysteine. MTT assay results revealed that ZPE (10-85 µ g/mL) could effectively inhibit the growth of AGS gastric cancer cells at higher concentrations (P<0.05, P<0.01). The annexin V & dead cell staining assay and cell cycle arrest assay confirmed a dose-dependent increase in the apoptosis rate and G@*CONCLUSION@#ZPE decreases AGS cell proliferation and induces apoptosis by inhibiting Akt and MDM2 expression.

Phenylpropanoids from Zanthoxylum species and their pharmacological activities: a review / 中国中药杂志

Phenylpropanoids are one of the major chemical constituents in Zanthoxylum species. They include simple phenylpropanoids, coumarins, and lignans and possess anti-tumor, anti-inflammatory, anti-platelet aggregation, anti-bacterial, anti-viral, insecticidal, and antifeedant activities. This review summarizes the chemical constituents and pharmacological activities from the Zanthoxylum plants in hopes of providing reference for the research and application of phenylpropanoids from this genus.

Biological activities and phytochemical screening of leaf extracts from Zanthoxylum caribaeum L. (Rutaceae) / Atividades biológicas e prospecção fitoquímica de extratos vegetais das folhas de Zanthoxylum caribaeum L. (Rutaceae)

The Brazilian flora is known for its vast biodiversity; however, many species have been still little studied regarding to their chemical composition and biological potential. Thus, this study aimed to determine the antimicrobial, antioxidant and acaricidal activity of the extracts of leaves of Zanthoxylum caribaeum L. In addition, phytochemical screening of these extracts was carried out to determine the main classes of secondary metabolites present in Z. caribaeum. Using the Z. caribaeum leaves, aqueous and organic extracts were obtained using the following solvents (ethanol, methanol, hexane, acetone, dichloromethane and ethyl acetate). The antimicrobial activity of extracts was determined by broth microdilution method, and to detect antioxidant activity the method of capturing the free radical 2,2-diphenyl-1-picryl hydrazyl (DPPH) was used. The acaricidal activity of the extracts was tested on Dermanyssus gallinae (De Geer) (Acari: Dermanissidae). Ethanolic and methanolic extracts presented antimicrobial activity for most of the bacterial strains tested, as well as for yeast Candida albicans. The ethanolic extract presented high free radical sequestration potential (71.2%) and antioxidant capacity (the lowest IC50 value - 24.39 µg mL-1). The crude extracts obtained with methanol and acetone were the most promising. In general, phytochemical screening indicated the presence of steroids, flavanones, flavones, flavonols, saponins, tannins, triterpenoids and xanthones.

A flora brasileira é conhecida pela sua vasta biodiversidade, no entanto, muitas espécies ainda são pouco estudadas quanto à composição química e ao potencial biológico. Assim, esse trabalho teve como objetivo determinar a atividade antimicrobiana, antioxidante e acaricida dos extratos vegetais das folhas de Zanthoxylum caribaeum L. Adicionalmente, foi realizada triagem fotoquímica desses extratos para determinar as principais classes de metabólitos secundários presentes em Z. caribaeum. Empregando-se as folhas de Z. caribaeum foram obtidos o extrato aquoso e orgânicos, utilizando os seguintes solventes (etanol, metanol, hexano, acetona, diclorometano e acetato de etila). A atividade antimicrobiana dos extratos foi determinada pelo método de microdiluição em caldo, e para detecção da atividade antioxidante foi empregado o método de captura do radical livre 2,2-difenil-1-picril hidrazil (DPPH). A atividade acaricida dos extratos foi avaliada frente a Dermanyssus gallinae (De Geer) (Acari: Dermanissidae). Os extratos brutos etanólico e metanólico apresentaram atividade antimicrobiana para a maioria das cepas bacterianas testadas, e também para a levedura Candida albicans. O extrato etanólico apresentou elevado potencial de sequestro de radicais livres (71,2%) e o menor valor de IC50 (24,39µg mL-1), revelando, portanto, sua capacidade antioxidante. No que se refere à atividade acaricida, os extratos obtidos com metanol e acetona foram os mais promissores. De modo geral, a triagem fitoquímica indicou a presença de esteroides, flavanonas, flavonas, flavonóis, saponinas, taninos, triterpenóides e xantonas.

Vasorelaxant activity and acute toxicity of the ethanolic extract of Zanthoxylum rhoifolium Lam leaves

The study evaluated the vasorelaxant effect induced by the ethanolic extract of the leaves of Zanthoxylum rhoifolium Lam (EEtOH-Zr/leaves). Wistar rats were treated with the leaf extract containing a single dose of 2,000 mg / kg, v.o. After 14 days, the animals were anesthetized for blood collection and subsequent analysis of the biochemical parameters; they were then euthanized (sodium pentobarbital-100 mg/kg, i.p.) for the removal and morphological analysis of the heart, lung, liver and kidney. The vasorelaxation activity the and vascular reactivity of EEtOH-Zr/leaves were evaluated on artery mesenteric rings isolated from rats. The extract showed no signs of toxicity and no significant difference in the values of the biochemical parameters between the control group and the group of treated animals. In the evaluation of pharmacological activity in the smooth muscle, the EEtOH-Zr/leaves caused vasorelaxant effect on the tonic contraction induced by phenylephrine in mesenteric artery preparations in the presence (pD2=2.17±0.05 µg/mL; Emax=99.8±5.2%) and absence (pD2=2.14±0.05 µg/mL; Emax=95.3±6.4%) of the vascular endothelium. Oral administration of EEtOH-Zr/leaves reduced the contraction induced by the cumulative addition of PHE. It is concluded that the EEtOH-Zr/leaves promote vasorelaxation and reduce vascular reactivity of adrenergic alpha-1 agonist in the mesenteric artery. The results did not show toxic effects of the extract.

Chemical and cytotoxic constituents of Zanthoxylum nitidum / 中国中药杂志

Ten compounds were isolated from the methanol extract of Zanthoxylum nitidum through silica gel, Sephadex LH-20, RP-18 and HPLC chromatography techniques. Their structures were elucidated by the MS and NMR spectra as zanthonitidine B(1), cyclo-(Leu-Leu-Leu-Leu-Ile)(2),6S-10-O-demethylbocconoline(3), liriodenine(4), isoplatydesmine(5), 5, 5'-dimethoxylariciresinol(6), syringaresinol (7), episyringaresinol (8), marmesin (9) and syringaldehyde (10). Among them,1 is a new alkaloid,2 is a cyclopentapeptide isolated from plant kingdom for the first time, and 3 is from the genus Zanthoxylum for the first time. Compounds 3 and 4 exhibited cytoxoxicity against three human cancer cell lines HT29, A549 and MDA-MB-231 with IC₅₈ values of 27.37, 24.10, 33.58 μmol·L⁻¹ and 9.12,6.05, 11.35 μmol·L⁻¹, respectively.

Anticancer properties and enhancement of therapeutic potential of cisplatin by leaf extract of Zanthoxylum armatum DC

BACKGROUND: Clinical use of chemotherapeutic drug, cisplatin is limited by its toxicity and drug resistance. Therefore, efforts continue for the discovery of novel combination therapies with cisplatin, to increase efficacy and reduce its toxicity. Here, we screened 16 medicinal plant extracts from Northeast part of India and found that leaf extract of Zanthoxylum armatum DC. (ZALE) induced cytotoxicity as well as an effect on the increasing of the efficiency of chemotherapeutic drugs (cisplatin, mitomycin C and camptothecin). This work shows detail molecular mechanism of anti-cancer activity of ZALE and its potential for combined treatment regimens to enhance the apoptotic response of chemotherapeutic drugs. RESULTS: ZALE induced cytotoxicity, nuclear blebbing and DNA fragmentation in HeLA cells suggesting apoptosis induction in human cervical cell line. However, the apoptosis induced was independent of caspase 3 activation and poly ADP ribose polymerase (PARP) cleavage. Further, ZALE activated Mitogen-activated protein kinases (MAPK) pathway as revealed by increased phosphorylation of extracellular-signal-regulated kinases (ERK), p38 and c-Jun N-ter-minal kinase (JNK). Inhibition of ERK activation but not p38 or JNK completely blocked the ZALE induced apoptosis suggesting an ERK dependent apoptosis. Moreover, ZALE generated DNA double strand breaks as suggested by the induction γH2AX foci formation. Interestingly, pretreatment of certain cancer cell lines with ZALE, sensitized the cancer cells to cisplatin and other chemotherapeutic drugs. Enhanced caspase activation was observed in the synergistic interaction among chemotherapeutic drugs and ZALE. CONCLUSION: Purification and identification of the bio-active molecules from the ZALE or as a complementary treatment for a sequential treatment of ZALE with chemotherapeutic drugs might be a new challenger to open a new therapeutic window for the novel anti-cancer treatment.

Research progress on alkaloids constituents from Zanthoxylum and their pharmacological activities / 中国中药杂志

There are 250 species of Zanthoxylum (Rutaceae) in the world. This genus distributed in tropical and subtropical regions. Alkaloids are the major and representative ingredients in these plants including quinolines, isoquinolines, and amide alkaloids, with such biological activities as anti-tumor, anti-inflammatory, analgesic, anti-virus, anti-platelet aggregation, anti-bacteria and anti- oxidant. These species have been used for a long time to treat toothache, urinary and venereal diseases, lumbago and rheumatism. This review summarizes the chemical constituents and pharmacological activities from the Z. sppplants, in an effort to the systematic research and application of the alkaloids of this genus.

A new dimeric lignan from Zanthoxylum simulans / 中国中药杂志

To study the chemical constituents from Zanthoxylum simulans and their anti-inflammatory activity. The constituents of Z. simulans were isolated and purified using various column chromatographies. Their chemical structures were elucidated using extensive spectroscopic methods. The compounds were assayed inhibitory activity against NO production in LPS stimulated RAW 264.7 cells. Four compounds were obtained from the ethanol extract of Z. simulans and determined to be isozanthpodocarpin B(1), kobusin (2), (+)-fargesin (3), and epieudesmin (4). Compound 1 exhibited NO production inhibitory effect with IC50 value of 14.49 µmol · L(-1). Compound 1 is a new dimeric lignan and may be serve as potential anti-inflammatory agent in the future.

Evaluation of the diuretic effects of crude stem bark extraction of Zanthoxylum heitzii (Rutaceae) in Wistar rats / 中西医结合学报

<p><b>OBJECTIVE</b>Zanthoxylum heitzii is a medicinal plant widely used in central Africa for the treatment of many diseases, especially cardiovascular diseases and hypertension. The diuretic effects of crude stem bark extraction were determined and its safety in rats was evaluated.</p><p><b>METHODS</b>The diuretic effects of crude stem bark extraction of Z. heitzii 250 g ± 10 g) of both sexes. The crude stem bark extraction of Z. heitzii at the doses of 225, 300 and 375 mg/kg was administered to rats at 5 mL/kg body weight. Urine volume was determined 1, 2, 3, 4, 5, 6 and 24 h after administration of the extract. Kinetics of electrolyte elimination in response to a single oral administration dose of acute treatment was measured. The experiments were performed under the same conditions with two synthetic pharmacological diuretics considered as reference (furosemide and hydrochlorothiazide). Urinary and plasma concentrations of sodium and potassium ions were determined using flame photometry. Concentrations of creatinine, urea, glucose, albumin and electrolytes in the plasma and urine samples were evaluated using a two-way digital bidirectional spectrophotometer. The osmolarity of plasma and urine samples was measured by cytometry using an osmometer. Aldosterone was measured by radioimmunoassay.</p><p><b>RESULTS</b>The plant extract accelerated the elimination of overloaded fluid and increased urine volume and the excretion of Na+, K+ and Cl- 24 h after administration (P<0.05). The increase in elimination of Na+, K+, and Cl- induced by caused alkalinization of the urine, and showed a strong inhibitory effect on carbonic anhydrase and saluretic. These effects were mainly observed at the dose of 375 mg/kg. At the maximum diuretic response, urinary osmolarity decreased significantly (P<0.05) when compared to controls. The stability of aldosterone level, the absence of correlation with the plasma levels of Na+, and increased clearance of free water in the animals treated with indicated that increased diuresis and natriuresis were tubular in origin. No significant (P>0.05) changes were observed in the body temperature of the animals.</p><p><b>CONCLUSION</b>The significant increase in urine volume 24 h after treatment followed a dose-response pattern. The excretion of Na+, K+ and Cl- caused a decrease in urine osmolarity. The stability of aldosterone, the absence of correlation with the plasma levels of sodium, and increased clearance of free water in animals treated with aqueous extract suggest that increased diuresis and moderate natriuresis elevation were of tubular origin.</p>

Bioactive Constituents from the Leaves of Zanthoxylum schinifolium

Activity-guided separation of the methylene chloride-soluble fraction of the leaves of Zanthoxylum schinifolium, resulted in the isolation of four coumarinoids (1 - 4), two triterpenoids (5, 6) and three fatty acid derivatives (7 - 9) as active principles. Their chemical structures were identified as collinin (1), 8-methoxyanisocoumarin (2), 7-(6'R-hydroxy-3',7'-dimethylocta-2',7'-dienyloxy)-coumarin (3), (E)-4-methly-6-(coumarin-7'-yloxy) hex-4-enal (4), lupeol (5), epi-lupeol (6), phytol (7), hexadec-3-enoic acid (8) and palmitic acid (9), on the basis of spectroscopic (1D, 2D and MS) data analyses and comparing with the data published in the literatures. Compounds 1 and 7 showed potent cytotoxicity against Jurkat T cells with IC50 values of 45.58 and 47.51 microM, respectively. The others showed moderate activity with IC50 values ranging around 80.58 to 85.83 microM, while the positive control, auraptene, possessed an IC50 value of 55.36 microM.

Topical application of herbal formula for the treatment of ligature-induced periodontitis

PURPOSE: The aim of this study was to investigate the therapeutic effects of a herbal formula, PerioH-035, containing Angelica sinensis, steamed Rehmannia glutinosa, Angelica dahurica, Cimicifuga heracleifolia, and Zanthoxylum piperitum on the periodontal breakdown in a well-established ligature-induced periodontitis model in rats. METHODS: Sprague-Dawley rats were randomly assigned to 1 of 4 groups: NL (non-ligatured), L (ligatured), P1 (ligatured and treated with 1 mg/mL PerioH-035), P100 (ligatured and treated with 100 mg/mL PerioH-035). Periodontitis was induced by placing a ligature around the mandibular first molars. PerioH-035 was topically applied to both sides of the first molar for 2 weeks. The right side of the mandibles was retrieved for micro-computed tomography (CT) and methylene blue staining to analyze alveolar bone loss. The left side of the mandibles was histologically analyzed by TRAP and H&E staining. The MMP-9 mRNA level in gingival tissue was investigated by RT-PCR. RESULTS: Alveolar bone resorption was significantly reduced in the PerioH-035-treated groups. The number of dense multi-nucleated cells found to be TRAP-positive by staining in the ligatured rats was markedly decreased by PerioH-035 application. In addition, periodontal tissue destruction, especially cementum demineralization, was ameliorated in the P1 and P100 groups. Moreover, gingival tissue from the PerioH-035-treated group showed a decrease in the MMP-9 mRNA level, resulting in recovery of collagen degradation. CONCLUSIONS: These results suggest that PerioH-035 has therapeutic effects on periodontitis, and thus, PerioH-035 shows promise as a treatment for periodontitis. GRAPHICAL ABSTRACT:

Avaliação da atividade antioxidante e alelopática do extrato etanólico e frações das cascas do caule de Zanthoxylum rhoifolium Lam., Rutaceae / Assessment of antioxidant and allelopathic activities of ethanolic extract and solvent fractions from stem bark of Zanthoxylum rhoifolium Lam. (Rutaceae)

The purpose of this study was to test for antioxidantand allelopathic activities in stem bark of Zanthoxylum rhoifolium Lam., Rutaceae, with the eventual aim of discovering biologically active substances. The plant material was subjected to ethanolic extraction and this extract was partitioned, yielding hexane, chloroform, ethyl acetate and hydroalcoholic fractions. Antioxidant activity was estimated by the reduction of phosphomolybdenum complex, of DPPH. and of thiobarbituric acid-reactive substances (TBARS). To detect allelopathy, the samples were tested, at four concentrations, on the germination and development of the radicle and hypocotyl of L. sativa seeds. The samples showed significant antioxidant activity against the reduction of the phosphomolybdenum complex, as compared to rutin, and reduction of TBARS, ascompared to BHT, as well as allelopathic activity, since they stimulated growth and seed germination. The chloroform and ethyl acetate fractions showed the best antioxidant potentials, with 204.17% and 127.11% compared to rutin, in the reduction of phosphomolybdenum complex, as did the crude ethanolic extract and hexane fraction, with 64.2% and 60.9% compared to BHT, in the TBARS method. In the allelopathic assay, the chloroform fraction stood out as the only sample that stimulated the growth of both the radicle and hypocotyl at most concentrations, ranging from 41 to 144%, while the ethyl acetate fraction achieved the greatest stimulus in this bioassay, increasing the growth of the hypocotyl by 274%. This is the first study that demonstrates the antioxidant and allelopathic activities of the species Z. rhoifolium...

Este trabalho teve como objetivo o estudo das atividades antioxidante e alelopática das cascas do caule de Zanthoxylum rhoifolium Lam., Rutaceae, de modo a conduzir à descoberta de substâncias biologicamente ativas. O material vegetal foi submetido à extração etanólica e este extrato foi fracionado obtendo as frações (hexano, clorofórmio, acetato de etila e hidroalcoólica). Para a avaliação da atividade antioxidante, empregaram-se os métodos de redução do complexo fosfomolibdênio, de redução do radical DPPH e das substâncias reativas ao acido tiobarbitúrico (TBARS). Quanto à alelopatia, as amostras foram testadas em quatro concentrações sobre a germinação e o desenvolvimento de radícula e hipocótilo das sementes de Lactuca sativa. As amostras evidenciaram atividade antioxidante significativa frente ao método de redução do complexo fosfomolibdênio quando comparada à rutina, e do TBARS quando comparado ao BHT, assim como a atividade alelopática, uma vez que estimularam tanto a germinação como o crescimento das sementes. A fração clorofórmica e acetato de etila demonstraram melhor potencial antioxidante com 204,17% e 127,11% em relação à rutina no método de formação do complexo fosfomolibdênio, e o extrato bruto e a fração hexano com 64,2% e 60,9%, em relação ao BHT, no método TBARS. No ensaio alelopático, destaca-se a fração clorofórmica, pois foi a única amostra que estimulou o crescimento do hipocótilo e radícula na maioria das concentrações, variando de 41 a 144%, e a fração acetato de etila que apresentou a maior porcentagem de estímulo nesse bioensaio, demonstrando estímulo de 274% do crescimento do hipocótilo. Este é o primeiro trabalho que demonstra a atividade antioxidante e alelopática de Z. rhoifolium...

Antibacterial, antifungal and antioxidant activities of some medicinal plants

The purpose of this study was to evaluate the antibacterial, antifungal and antioxidant activities of medicinal plants. The antibacterial activity of methanolic extracts of three medicinal plants [Swertia chirata, Terminalia bellerica and Zanthoxylum armatum] were tested against Gentamicin [standard drug] on eleven gram positive and seventeen gram negative bacteria by agar well method. It was revealed that seven-gram negative and six gram positive bacterial species were inhibited by these plant extracts. Minimum inhibitory concentrations [MIC] of the extracts were determined by broth micro-dilution method. The significant MIC value of Swertia chirata was 20mg/ml against Serratia marcesens, Zanthoxylum armatum was 10 mg/ml against Aeromonas hydrophila and Terminali bellerica was 20mg/ml against Acinetobacter baumanii as well as Serratia marcesens. Antifungal screening was done for methanolic extracts of these plants by agar well method with the 6 saprophytic, 5 dermatophytic and 6 yeasts. In this case Griseofulvin was used as a standard. All saprophytes and dermatophytes were showed resistance by these plants extracts except Microsporum canis, which was inhibited by Z. armatum and S. chirata extracts. The significant MIC value of Zanthoxylum armatum was 10mg/ml against Microsporum canis and Swertia chirata was 10mg/ml against Candida tropicalis. The anti-oxidant study was performed by DPPH free radical scavenging assay using ascorbic acid as a reference standard. Significant antioxidant activities were observed by Swertia chirata and Zanthoxylum armatum at concentration 200microg/ml was 70% DPPH scavenging activity [EC[50]=937.5microg/ml] while Terminalia bellerica showed 55.6% DPPH scavenging activity [EC[50]=100ug/ml]. This study has shown that these plants could provide potent antibacterial compounds and may possible preventive agents in ROS related ailments

Fruits and barks extracts of Zanthozyllum heitzii a spice from Cameroon induce mitochondrial dependent apoptosis and Go/G1 phase arrest in human leukemia HL-60 cells

BACKGROUND: Zanthoxylum heitzii is a spice used to prepare several dishes and to treat tumors, syphilis, malaria, cardiac palpitations, urogenital infections in the west region of Cameroon, but the antitumor mechanisms and chemical composition are not yet investigated. This study was aimed to determine the antiproliferative effects of four extracts from the fruits and barks of Zanthoxyllum heitzii (Rutaceae) on apoptosis in human promyelocytic cells, their mechanisms and the chemical composition. The 3-(4, 5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the fifty percent inhibition (IC50) concentration of the cell lines after treatment. The effect on morphology was observed using a light or fluorescence microscopy. The rate of apoptosis and the cell cycle were measured using flow cytometry (FCM). The phytochemical analysis of the extract was carried with HPLC/MS methods. RESULTS: The phytochemical analysis of the extracts indicated the presence of four known polyphenols (Syringic acid, Juglon, Luteolin and Myricetin) in both fruits and barks of Z. heitzii but in different quantities. Syringic acid and Myricetin concentrations were between 17-21 fold higher in the fruits than the stem bark. Rhamnetin (393.35 µg/mL) and Oleuropein (63.10 µg/mL) were identified only in the stem barks of Z. heitzii. Among the four extracts tested for cytotoxicity properties, only the methanol extract of fruits and barks significantly inhibited cell proliferation of HL-60 cells with IC50 value of 20 µg/mL and 12 µg/mL respectively. HL-60 cells treated with Z. heitzii extracts significantly produced reactive oxygen species (ROS) with concurrent loss of mitochondrial membrane potential (MMP). Modifications in the DNA distribution and enhanced of G1/G0 phase cell cycle arrest were observed in a concentration dependent manner. CONCLUSIONS: Polyphenols from Z. heitzii plant exert inhibitory effect on HL-60 cells through the reactive oxygen species (ROS) generation, loss of mitochondrial membrane potential and cell cycle destabilization.

GC-MS analysis and cytotoxic activity of the supercritical extracts from roots and stems of Zanthoxylum nitidum / 中国中药杂志

The volatile components of roots and stems of Zanthoxylum nitidum were investigated by supercritical fluid carbon dioxide extraction (SFE-CO2) and gas chromatography-mass spectrometry(GC-MS). Thirty-one and fifty-one compounds were identified in the supercritical extracts from roots and stems of Z. nitidum, respectively, and total twenty-seven compounds were the common constituents. Among them, the major constituents in root and stem supercritical extracts were spathulenol (18.49 and 26.18%), n-hexadecanoic acid (14.24% and 12.79%), ar-tumerone (6.95% and 8.88%), oleic acid (8.39% and 5.71%) and hexanoic acid (4.39% and 7.78%). The in-vitro MTT assay showed that the volatile components of roots and stems of Z. nitidum did not exhibited any cytotoxic activity against human cancer Huh-7 and normal IEC-6 cells. These results indicated the same nature of the volatile constituents in the root and stem of Z. nitidum. This investigation may provide further evidence for expansion of medicinal parts of Z. nitidum.

Efecto in vitro de los terpenos lupeol y casearina G sobre células sanguíneas y tumorales / In vitro effect of lupeol and casearin G on peripheral blood mononuclear and tumor cells

Background: The rainforest is an important source of natural compounds with therapeutic properties. Although there are many anti-inflammatory and antineoplastic drugs available to the clinician, there is an ongoing need for new therapeutic drugs with fewer serious adverse effects. Aim: To evaluate the in vitro cytotoxic effects of lupeol and casearin G on tumor cells, on phagocytic activity and nitric oxide (NO) production by blood mononuclear cells. Material and Methods: The cytotoxic effect of these compounds on cell lines MCF-7 (human breast adenocarcinoma) and PC-3 (human prostate cancer) was measured by a colorimetric assay (MTS/PMS) and the sulphorhodamine B assay. Peripheral blood mononuclear cells were obtained from eight healthy volunteers. The effect of these compounds on nitric oxide (NO) production was measured using the Griess reaction. Their effect on phagocytic activity of PBMC was also evaluated. Results: Lupeol (≥ 2 mM) resulted in a reduction of both the phagocytic index and the percentage of phagocytic monocytes and macrophages. Treatment of monocytes/macrophages with lupeol (72 µM) and casearin G (4 µM) reduced the production of NO. Neither lupeol (< 969 µM) nor casearin G (< 55 µM) had cytotoxic effects on PBMC. Casearin G showed both cytotoxic (IC50, LC50) and cytostatic (GI50) effects against tumor cells, PC-3 (IC50 = 12.5 µM; GI50 = 13.3 µM; LC50 = 51.9 µM) and MCF-7 (IC50 = 112.8 µM; GI50 = 11.8 µM; LC50 = 49.4 µM), as well as a hemolytic effect (≥ 182 µM). Conclusions: These observations indicate that lupeol and casearin G might be useful compounds in the preparation of anti-inflammatory drugs, whereas casearin G might be useful in the elaboration of antitumor drugs.

Antihypertensive and vasorelaxant effects of ethanol extract of stem barks from Zanthoxylum rhoifolium Lam in rats.

Administration of ethanol extract of stem bark from Z. rhoifolium (EEtOH-ZR) induced hypotension associated with a dual effect in heart rate in normotensive rats. This response was highlighted in spontaneously hypertensive rats (SHR). In rat superior mesenteric artery rings, the cumulative addition of EEtOH-ZR (0.1–750 µg/mL) on a phenylephrine-induced pre-contraction (10-5 M) promoted a vasorelaxant effect by a concentration-dependent manner and independent of vascular endothelium. A similar effect was obtained on KCl-induced pre-contractions (80 mM). EEtOH-ZR attenuated contractions induced by cumulative addition of CaCl2 (10-6–3 × 10-2 M) in depolarizing medium without Ca2+ only at 500 or 750 µg/mL. Likewise, on S-(–)-Bay K 8644-induced pre-contractions (10-7 M), the EEtOH-ZR-induced vasorelaxant effect was attenuated. EEtOH-ZR (27, 81, 243 or 500 µg/mL) inhibited contractions induced by cumulative addition of phenylephrine (10-9 - 10-5 M) in endothelium-denuded preparations or by a single concentration (10-5 M) in a Ca2+-free medium. The involvement of K+ channels was evaluated by tetraethylammonium (3 mM); the EEtOH-ZR-induced vasorelaxation was not attenuated. Thus, calcium influx blockade through voltage-operated calcium channels (CaVL) and inhibition of calcium release from intracellular stores are probably underlying EEtOH-ZR-induced cardiovascular effects.

Analysis of alkaloids in Zanthoxylum nitidum by HPLC-DAD/ESI-Q-TOF-MS / 中国中药杂志

The alkaloids in Zanthoxylum nitidum were identified by HPLC-DAD/ESI-Q-TOF-MS. Separation was performed on a Hanbon C18 column with acetonitrile (with 0.1% formic acid) and water(with 0.1% formic acid) as mobile phase. Based on the high-resolution mass information, MS/MS fragmentation behaviors and chemical components from literatures, 48 components were identified or tentatively characterized including 6 new compounds. This work could be useful for the quality control and further studies of the plant.