Zanthoxylum Piperitum DC Research Articles

Classification of Japanese Pepper (Zanthoxylum piperitum DC.) from Different Growing Regions Based on Analysis of Volatile Compounds and Sensory Evaluation.

The Japanese pepper (Zanthoxylum piperitum DC.) is an attractive plant that is highly palatable and benefits human health. There are several lineages of pepper plants in Japan. However, the classification of each lineage by analyzing its volatile compounds and studies on the effects of differences in volatile compounds on human flavor perception have not been performed in detail. Herein, we conducted gas chromatography (GC) and GC/mass spectrometry (GC/MS) analysis of volatile compounds and sensory evaluation of flavor by an analytical panel using 10 commercially available dry powdered Japanese pepper products from different regions. GC and GC/MS analysis detected limonene, β-phellandrene, citronellal, and geranyl acetate as the major volatile compounds of Japanese peppers. The composition of volatile compounds showed different characteristics depending on the growing regions, and cluster analysis of composition classified the products into five groups. The sensory evaluation classified the products into four groups, and the results of both classifications were in good agreement. Our results provide an important basis for proposing cooking and utilization methods that take advantage of the unique characteristics of each lineage based on scientific evidence.

Skin care benefits of bioactive compounds isolated from Zanthoxylum piperitum DC. (Rutaceae)

Purpose: To investigate skin care efficacies of Zanthoxylum pipetitum extract and isolated compounds. Methods: Ethanol extracts of leaves, branches and fruits of what were partitioned into n-hexane, chloroform, ethyl acetate, n-butanol and aqueous layers and some fractions were further analyzed to isolate five compounds. The isolated compounds were identified based on the proton and carbon nuclear magnetic resonance (NMR) spectra. Cosmetic efficacy tests of the extracts and isolated compounds were evaluated by in vitro tests. Results: Phytochemical studies of the chloroform and ethyl acetate layers led to the isolation of five compounds; quercitrin (1), afzelin (2), hydroxy-α-sanshool (3), α-sanshool (4) and hyperoside (5). In activity tests, the extracts showed inhibitory activity against inflammation response and melanin synthesis, and induction of procollagen type I C-peptide (PIP). Among the isolated compounds, hydroxy-α-sanshool (3) and α-sanshool (4) displayed significant anti-inflammatory activity. Conclusion: The results demonstrate that Z. piperitum extract and its active compounds possess a significant potential as a cosmeeutical agent for enhancing skin quality.

産地の異なる山椒（Zanthoxylum piperitum DC.）の香気成分分析と官能評価による分類

産地の異なる山椒（Zanthoxylum piperitum DC.）の香気成分分析と官能評価による分類

초피 추출물을 이용한 친환경 항균 실내 직물 블라인드 소재 개발

본 연구는 초피의 항균 활성 소재의 산업화 타진 연구로 초피과피 정유 성분의 경우 높은 생산단가로 인해 그 활용이 제한적이다. 이에 초피잎을 대상으로 항균 활성소재를 탐색하고 초피의 천연 방향성과 항균력을 이용하여 실내 환경 개선 소재 특히 친환경 항균블라인드 개발에 대한 활용 가능성을 타진하였다. 그 결과, 초피잎 추출물의 항균 활성은 초피과피 추출물보다 약하게 나타났지만, K. pneumoniae, S. aureus와 S.mutans 균에 대하여 IC₅₀ 값은 0.5 mg/ml, 0.1 mg/ml와 1.0 mg/ml로 운향과 유래 타 Zanthoxylum 속 보다 항균력이 강한 것으로 나타났다. 항균력이 강하게 확인된 초피잎 EtOH 추출물과 EtOAc 추출물의 GC-MS 분석을 통해 초피잎 유래의 항균력 관련 화합물로서 quinic acid, 3,5-dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one과 kaempferol 등을 확인할 수 있었다. 또한, International Standard Fabrics Test Method에 준하여 초피 추출물을 처리한 블라인드 소재의 항균력을 조사한 결과, 1% 농도의 초피잎 EtOH 추출물을 처리한 직물 블라인드 소재의 경우는 항생제 내성균 S. aureus의 생육을 99.9% 저해하였고, 2% 농도의 처리에 의해서는 K. pneumoniae 생육도 완전히 저해하였다. 이러한 연구결과는, 고가인 초피과피에 비해 농산폐자원으로 폐기되고 있는 초피잎의 항균력을 이용하여 친환경 기능성 항균 직물 블라인드 및 실내 창호 소재를 개발한다면 최근 급증하는 실내 환경성 질환의 예방 및 제어에 매우 유용할 수 있음을 시사한다. As the incidence of environmental diseases is increasing due to harmful environmental factors, there is a rising interest in developing eco-friendly materials for housing. In this study, we sought to develop antimicrobial interior fabric blind materials by employing ethanol extract of a medicinal plant Zanthoxylum piperitum DC. As determined by the disc diffusion method, the zones of inhibition of the pericarp ethanol extract at a concentration of 5 mg/disc against Klebsiella pneumoniae, Staphylococcus aureus, and Streptococcus mutans were 13.5±1.5 mm, 14.0±0.5 mm and 15.0±0.1 mm, respectively, whereas the leaf ethanol extract (5 mg/disc) against K. pneumoniae, S. aureus, and S. mutans were 12.8± 0.3 mm, 13.5±1.0 mm, and 12.0±0.1 mm, respectively. The IC₅₀ of the leaf ethanol extract against K. pneumoniae, S. aureus and S. mutans were about 0.5 mg/ml, 0.1 mg/ml and 1.0 mg/ml respectively. To examine whether the leaf ethanol extract possessing antibacterial activity of Z. piperitum DC can be applicable to production of antimicrobial fabric blind materials, the fabrics treated with either 1.0% or 2.0% of the leaf ethanol extract were tested for antibacterial activity against K. pneumoniae and S. aureus using International Standard Fabrics Test Method. The results indicate that the fabric treated with the ethanol extract of Z. piperitum DC possesses an excellent antimicrobial activity against both pathogenic bacteria. These results suggest that Z. piperitum DC may be applicable to producing functional fabrics which are effective in reducing the harmful bacterial factors in indoor environments.

Antinociceptive Activity of Zanthoxylum piperitum DC. Essential Oil.

Zanthoxylum piperitum DC. (ZP) is a traditional medicinal plant used mainly in countries from Asia such as Japan. This study aimed to investigate the antinociceptive effect of ZP essential oil (ZPEO). The major component present in the essential oil was beta-phellandrene (29.39%). Its antinociceptive activity was tested through animal models (formalin-, capsaicin-, and glutamate-induced paw licking and hot plate). The anti-inflammatory effect was evaluated through the carrageenan-induced leukocyte migration into the subcutaneous air pouch (SAP), with measurement of cytokines. The results showed antinociceptive effect for ZPEO for the first phase of the formalin-induced licking, glutamate, and hot plate tests. However, ZPEO had no effect on reducing paw licking induced by capsaicin. Finally, ZPEO had no effect against inflammation induced by carrageenan.

Expression of IPT in Asakura-sanshoo (Zanthoxylum piperitum (L.) DC. f. inerme Makino) Alters Tree Architecture, Delays Leaf Senescence, and Changes Leaf Essential Oil Composition.

The IPT gene encodes isopentenyl pyrophosphate transferase, a key enzyme in cytokinin biosynthesis. We introduced IPT under the control of the CaMV35S promoter into Asakura-sanshoo (Zanthoxylum piperitum (L.) DC. f. inerme Makino) via stable Agrobacterium tumefaciens-mediated transformation. Three of 3-year-old transgenic Asakura-sanshoo lines Y5, Y16, and Y17 were used to evaluate the effects of IPT expression on the morphological characteristics, leaf senescence, and essential oil composition. Introduced IPT into Asakura-sanshoo stimulated an increase in cytokinin content and a decrease in auxin level. The increase in the cytokinin/auxin ratio affected the tree architecture in 3-year-old transgenic lines. The phenotypes of transgenic lines included reduced stem elongation, decreased leaf surface area, increased branching, and delayed leaf senescence. The expression of IPT in Asakura-sanshoo also affected the leaf essential oil composition. The amount of oxygenated sesquiterpenoid compounds in Y5 and Y16 was 21.1 and 15.8 % higher, respectively, than that in wild type (WT). The amount of aromatic compounds in Y5 and Y16 was 2.9 and 24.6 % lower, respectively, than that in WT. These results show that ipt expression in Asakura-sanshoo conferred desirable traits, including a dwarf growth habit, delayed senescence, and increased concentrations of some sesquiterpenoid compounds.Electronic supplementary materialThe online version of this article (doi:10.1007/s11105-015-0948-9) contains supplementary material, which is available to authorized users.

Bioactivity of proteins isolated from Lactobacillus plantarum L67 treated with Zanthoxylum piperitum DC glycoprotein

Lactobacilli in the human gastrointestinal tract have beneficial effects on the health of their host. To enhance these effects, the bioactivity of lactobacilli can be fortified through exogenous dietary or pharmacological agents, such as glycoproteins. To elucidate the inductive effect of Zanthoxylum piperitum DC (ZPDC) glycoprotein on Lactobacillus plantarum L67, we evaluated the radical-scavenging activity, anti-oxidative enzymes (SOD, GPx and CAT), growth rate, ATPase activity and β-galactosidase activity of this strain. When Lact. plantarum L67 was treated with ZPDC glycoprotein at different concentrations, the intensities of a few SDS-PAGE bands were slightly changed. The amount of a 23 kDa protein was increased upon treatment with increasing concentrations of ZPDC glycoprotein. The results of this study indicate that the radical-scavenging activity for O2(-) and OH¯, but not for the DPPH radical, increased in a concentration-dependent manner after treatment with ZPDC glycoprotein. The activation of anti-oxidative enzymes (SOD, GPx and CAT), growth rate and β-galactosidase activity also increased in a concentration-dependent manner in response to ZPDC glycoprotein treatment, whereas ATPase activity was decreased. In summary, ZPDC glycoprotein stimulated an increase in the bioactivity of Lact. plantarum L67. Significance and impact of the study: This study demonstrated that Lactobacillus plantarum L67 possesses anti-oxidative activity. This strain of lactic bacteria has been known to have various probiotic uses, such as yogurt starters and dietary additional supplements. We found, through this experiment, that the protein has a strong anti-oxidative character, and the activity can be enhanced by treatment with Zanthoxylum piperitum DC (ZPDC) glycoprotein. This study may be application of Lact. plantarum L67 treated by ZPDC glycoprotein in yogurt fermentation. It could be one of the avenues of minimizing yogurt postacidification during storage. In addition, it can be manufactured and incorporated in food products without losing viability and functionality of Lact. plantarum L67.

올리고당의 이해

This study was to investigate whether the bioactivity of L. plantarum L67 can be enhanced by treatment with Zanthoxylum piperitum DC (ZPDC) glycoprotein or compounds of similar molecular weights (24 kDa). When L. plantarum L67 was treated with ZPDC glycoprotein of various concentrations, the intensities of a few SDS-PAGE bands were slightly changed. The amount of a 23-kDa protein was increased upon treatment with increasing concentrations of ZPDC glycoprotein. The activities of antioxidant enzymes (SOD, GPx, and CAT), growth rate, and β-galactosidase activity were more increased at treatment with ZPDC glycoprotein than at treatment of L. plantarum L67 only, whereas the ATPase activity was decreased. Interestingly, it did not affect scavenging activity for the DPPH. The founding result suggest that, one of beneficial strategies for increase the bioactivity of L. plantarum L67 might be to take ZPDC.

Screening and identification of phytotoxic volatile compounds in medicinal plants and characterizations of a selected compound, eucarvone.

Screening and identification of phytotoxic volatile compounds were performed using 71 medicinal plant species to find new natural compounds, and the characterization of the promising compound was investigated to understand the mode of action. The volatile compounds from Asarum sieboldii Miq. showed the strongest inhibitory effect on the hypocotyl growth of lettuce seedlings (Lactuca sativa L.cv. Great Lakes 366), followed by those from Schizonepeta tenuifolia Briquet and Zanthoxylum piperitum (L.) DC.. Gas chromatography-mass spectrometry (GC/MS) identified four volatile compounds, α-pinene (2,6,6-trimethylbicyclo[3.1.1]hept-2-ene), β-pinene (6,6-dimethyl-2-methylenebicyclo[3.1.1]heptane), 3-carene (3,7,7-trimethylbicyclo[4.1.0]hept-3-ene), and eucarvone (2,6,6-trimethy-2,4-cycloheptadien-1-one), from A. sieboldii, and three volatile compounds, limonene (1-methyl-4-(1-methylethenyl)-cyclohexene), menthone (5-methyl-2-(propan-2-yl)cyclohexan-1-one), and pulegone (5-methyl-2-propan-2-ylidenecyclohexan-1-one), from S. tenuifolia. Among these volatile compounds, eucarvone, menthone, and pulegone exhibited strong inhibitory effects on both the root and shoot growth of lettuce seedlings. Eucarvone-induced growth inhibition was species-selective. Cell death, the generation of reactive oxygen species (ROS), and lipid peroxidation were induced in susceptible finger millet seedlings by eucarvone treatment, whereas this compound (≤158μM) did not cause the increase of lipid peroxidation and ROS production in tolerant maize. The results of the present study show that eucarvone can have strong phytotoxic activity, which may be due to ROS overproduction and subsequent oxidative damage in finger millet seedlings.

Preventive effects of ZPDC glycoprotein (24 kDa) on hepatotoxicity induced by mercury chloride in vitro and in vivo.

Mercury is a potent environmental contaminant that exerts toxic effect on various vital organs in the human body. Recently, we isolated glycoprotein from Zanthoxylum piperitum DC (ZPDC), which has antioxidant and anticancer effects. In the present study, we determined the preventive effects of ZPDC glycoprotein on hepatic damage induced by mercury chloride (HgCl2 ). We evaluated the activities of lactate dehydrogenase (LDH), alanine aminotransferase (ALT), antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], extracellular signal-regulated kinase (ERK)1/2, p38 mitogen-activated protein kinase (MAPK), cyclo-oxygenase (COX-2), inducible nitric oxide synthetase (iNOS), and activator protein (AP-1) and the quantitative expressions of nuclear factor E2-related factor (Nrf2), heme oxygenase (HO-1), metallothionein (MT) and reduced glutathione (GSH) in mercury-chloride-exposed (50 μM and 10 mg/kg body weight) primary cultured hepatocytes and ICR mice, using biochemical assays, radioactivity and immunoblot analysis. The results demonstrated that ZPDC glycoprotein decreased the levels of LDH, ALT, HO-1 and MT, whereas it increased the activities of hepatic antioxidant enzymes (SOD, CAT and GPx) and reduced GSH in mercury-chloride-exposed primary cultured hepatocytes. Also, it suppressed arachidonic acid release and expression of ERK, p38 MAPK, COX-2, iNOS, AP-1 and Nrf-2 in primary cultured hepatocytes and ICR mice exposed to mercury chloride. Collectively, ZPDC glycoprotein may have potential applications to prevent hepatotoxicity induced by mercury chloride.

생강나무(Lindera obtusiloba Blume)와 초피나무(Zanthoxylum piperitum DC) 추출물의 항균활성

Ethanol and hot water extracts were prepared from Lindera obtusiloba Blume (LO) and Zanthoxylum piperitum DC (ZP) and used to evaluate their antimicrobial activities and thermal stability against six foodborne pathogens (3 gram-positive and 3 gram-negative bacteria). The antimicrobial activities were assessed using the agar diffusion method, and the thermal stabilities of extracts were examined after heat treatment at 60, 70, 80, and 100°C for 10 min. The zones of inhibition by the LO extract or the ZP extract of the tested microorganisms were in the range of 21-30 mm and 19-25 mm, respectively, at 100 mg/mL concentrations. The 60% ethanol extract and the hot water extracts from LO showed the strongest antimicrobial effects against MRSA and Staphylococcus aurues, respectively. For the extract from ZP, the strongest antimicrobial effect was shown against S. aurues by 60% ethanol, and the weakest antimicrobial effect was shown against E. coli by the hot water extracts. The ZP extracts showed that the gram-positive bacteria were more sensitive than gram-negative bacteria. For the thermal stability of the extracts, the antimicrobial effects stabilized after heat treatment. Overall, the data suggest that the extracts have a potential for application in various food products for which a natural antimicrobial additive is desired.

Antiviral effect of flavonol glycosides isolated from the leaf of Zanthoxylum piperitum on influenza virus

The ethanol extract of Zanthoxylum piperitum (L.) DC. showed in vitro antiviral activity against influenza A virus. Three flavonol glycosides were isolated from the EtOAc fraction of Z. piperitum leaf by means of activity-guided chromatographic separation. Structures of isolated compounds were identified as quercetin 3-O-β-D-galactopyranoside (1), quercetin 3-O-α-L-rhamnopyranoside (2), kaempferol 3-O-α-L-rhamnopyranoside (3) by comparing their spectral data with literature values. The anti-influenza viral activity of isolates was evaluated using a plaque reduction assay against influenza A/NWS/33 (H1N1) virus. The compounds also were subjected to neuraminidase inhibition assay in influenza A/NWS/33 virus. Compounds 1-3 exhibited antiviral activity against an influenza A virus in vitro, and inhibited the neuraminidase activity at relatively high concentrations.

ZPDC glycoprotein (24 kDa) induces apoptosis and enhances activity of NK cells in N-nitrosodiethylamine-injected Balb/c

Natural killer (NK) cells have anti-tumor activity in hepatocellular carcinoma (HCC) using secreting granules and cytotoxic ability. Recently, we isolated glycoprotein from Zanthoxylum piperitum DC (ZPDC) has anti-oxidant effect and anti-cancer effect. The objective of this study was to determine whether ZPDC glycoprotein enhances activity of NK cells and induces apoptosis of liver cancer cells in diethylnitrosamine (DEN)-treated Balb/c mice. This study evaluated the secreting of perforin and granzyme B and cytotoxicity of NK cells, interleukin (IL)-2 and IL-12, apoptosis-related factors (bid, cytochrome c, and caspase-3) in liver tissue using Immunoblot and ELISA. The results demonstrated that ZPDC glycoprotein (20mg/kg, BW) induces secretion of perforin and granzyme B and NK cells activity. Also, it induces expression of apoptosis-related factors (bid, cytochrome c, and caspase-3) in liver tissues. Collectively, ZPDC glycoprotein may have potential applications to prevent hepatocarcinogenesis without immunosuppression.

First Report of Alternaria alternata Causing Blight on Zanthoxylum piperitum in China.

Since 2009, brown leaf spot and panicle blight of Zanthoxylum piperitum (L.) DC. ("liujin," commonly known as Japanese pepper and Japanese pricklyash) has been observed on 40% of the plants in the test field of Foresty Academy of Science in Hebei Province of China. When symptoms formed on leaves, a thick yellow spot appeared, which then turned brown. When on the spikes, brown lesions were observed initially on the grain, which then spread down to fruit stem, and finally the whole spike wilted and dried up. Yield and quality losses were considerable. A fungus was isolated consistently from the diseased tissues using potato dextrose agar (PDA) (1). Three representative isolates were chosen for further characterization. All the isolates grew at 28°C on PDA and potato carrot agar (PCA) medium. Fungal colonies were initially white, then became olivaceous with some white mycelium on the top of the colony, and turned brown with age. When observed with the microscope, crineous septate hypha appeared, and conidiophore peduncles were upright or slightly curved, with a few branches, 33.0 to 75.0 μm long and 4.0 to 5.5 μm wide. Conidia were crineous short clubs or near oval in shape, 22.5 to 40.0 μm long and 8.0 to 13.5 μm wide, with a short conical beak, and had one to four longitudinal cross walls. On PCA, condia had three to seven transepta and one to five longisepta, and were produced in a branched, long chain with more than five conidia. The pathogen was identified based on morphological characteristics as Alternaria alternata (Fr.:Fr.) Keissl. (3). DNA was extracted from mycelium and PCR was performed on the internal transcribed spacer (ITS) region with primers ITS1 and ITS4. A 570-bp fragment was amplified and sequenced (GenBank Accession No. JQ973810). BLASTn analysis revealed there was 100% sequence identity with A. alternata strains (GU566303 and GQ121322). To further identify the fungus, A. alternata species-specific primers AAF2/AAR3 (2) were used to generate an amplicon which was then sequenced (JX308287). Sequence comparison showed there was 100% sequence identity with A. alternata (JQ927300 and JQ907485). Pathogenicity tests were performed by spraying with a cultured suspension (106 spores/ml) of approximately 100 μl onto healthy leaves in 15-cm-diameter glass dishes containing sterilized filter paper soaked with sterilized water at room temperature. Control plants were inoculated with sterile distilled water. Ten days after inoculation, symptoms were observed in all inoculated leaves and appeared to be identical to those observed in the field. No symptoms were noted on the control leaves. Identical results were also obtained when spikes were inoculated. The fungi reisolated from symptomatic plants were A. alternata. To our knowledge, this is the first report of A. alternata causing leaf spots and panicle blight of Z. piperitum in China.

Growth and Yield of ‘Budousanshou’ Grafts on ‘Fuyuzanshou’ and ‘Karasuzanshou’ Rootstocks

The growth and yield of ‘Budousanshou’ (Zanthoxylum piperitum (L.) DC. f. inerme Makino) trees were compared between grafts on ‘Fuyuzanshou’ (Z. alatum Roxb. var. planispinum Rehd. et Wils.) seedlings and ‘Karasuzanshou’ (Fagara ailanthoides Engl.) seedlings. Comparisons were made for 8 years after transplanting. The tree height for grafts on ‘Karasuzanshou’ rootstock was greater than for ‘Fuyuzanshou’ grafts throughout the experimental period after transplanting. For both graft treatments, tree height did not increase beyond 6–7 years after transplanting, suggesting that the trees had reached maturity. In addition, the canopy volume of grafts on ‘Karasuzanshou’ rootstock was greater than that on ‘Fuyuzanshou’ rootstock at 2 years after transplanting. The yield per tree and canopy volume were higher for grafts on ‘Karasuzanshou’ than grafts on ‘Fuyuzanshou’ rootstock by 7 years post-transplanting (i.e., the mature stage). Due to its greater canopy volume, the per tree yield of mature ‘Karasuzanshou’ grafts has the potential to be greater than that from ‘Fuyuzanshou’; therefore, ‘Karasuzanshou’ is a good candidate rootstock for the effective production of Japanese peppers.

Effects of the Timing of Fertilizer Application on the Number of Flowers of Japanese Pepper

This study was conducted in three consecutive years to determine the appropriate timing of fertilizer application to increase the flower number of the Japanese pepper, Zanthoxylum piperitum (L.) DC. f. inerme Makino (known as ‘Budousanshou’ in Japan) cultivated in Wakayama Prefecture. The timing experiment involved three treatment options: fertilizer application in spring with two equal applications in May and June, a summer/autumn treatment of two equal applications in August and September, or a spring/summer/autumn treatment of four equal applications in May, June, August, and September. Each treatment consisted of an annual application of 300 kg·ha−1 nitrogen. The spring treatment resulted in increased flower number. It was suggested that the application of fertilizer in the May/June spring treatment resulted in a focused accumulation of nutrients prior to the floral evocation period and led to the subsequent increase in flower number and cluster number relative to the summer/autumn and spring/summer/autumn treatments.

Protection Against Cyclophosphamide-Induced Myelosuppression by ZPDC Glycoprotein (24 kDa)

Immunomodulatory agents are often used to reduce myelosuppression and enhance immune response for cancer treatment. Cyclophosphamide (CTX) can induce oxidative stress in bone marrow resulting in suppression of anti-oxdiantive enzymes and causes myelosuppression. We isolated glycoprotein from Zanthoxylum piperitum DC fruit (ZPDC), and it consists of a carbohydrate (18%) and a protein (82%). The objective of this study was to investigate its protective activity against CTX-induced myelosuppression in Balb/c (n=6/group). The mice were orally administrated by ZPDC glycoprotein (10 and 20 mg/kg, BW) for 1 week in the presence or absence of CTX. Intracellular reactive oxygen species (ROS), anti-oxidant enzymes [superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT)], cyclin kinase inhibitors (CKIs: p53, p21 and p27), cyclin D1/ cyclin dependent kinase (CDK) 4, PCNA and cytokines [interleukin (IL)-3, and granulocyte⁄ macrophage-colony-stimulating factor (GM-CSF)] were evaluated using biochemical activity, Western blot analysis, and ELISA. The results obtained from this study showed that CTX decreased spleen and thymic indices, bone marrow cellularity and expression of cyclin D1/CDK4 and PCNA, but it increased CKIs, whereas ZPDC glycoprotein (20 mg/kg, BW) resulted in vice versa in CTX-induced Balb/c. Expression of IL-3 and GM-CSF were normalized by ZPDC glycoprotein. Thus, this study suggested that ZPDC glycoprotein prevents oxidative stress and myelosuppression in CTX-induced mice and might be a potential immunomodulatory agent.

Zanthoxylum piperitum DC Ethanol Extract Suppresses Fat Accumulation in Adipocytes and High Fat Diet-Induced Obese Mice by Regulating Adipogenesis

This study was conducted to determine the anti-obesity effects of Zanthoxylum piperitum DC fruit ethanol extract (ZPE) in 3T3-L1 adipocytes and obese mice fed a high-fat diet. We evaluated the influence of the addition of ZPE to a high-fat diet on body weight, adipose tissue weight, serum and hepatic lipids in C57BL/6 mice. In addition, adipogenic gene expression was determined by Western blot and real-time reverse transcription-PCR analysis. We assessed the effect of ZPE on 3T3-L1 preadipocyte differentiation. ZPE reduced weight gain, white adipose tissue mass, and serum triglyceride and cholesterol levels (p<0.05) in high-fat diet-fed C57BL/6 mice. ZPE decreased lipid accumulation and PPARγ, C/EBPα, SREBP-1, and FAS protein and mRNA levels in the liver. ZPE inhibited in vitro adipocyte differentiation in a dose-dependent manner and significantly attenuated adipogenic transcription factors, such as PPARγ, C/EBPα, and SREBP-1 in 3T3L1 cells. These findings suggest that Z. piperitum DC exerts an anti-obesity effect by inhibiting adipogenesis through the downregulation of genes involved in the adipogenesis pathway.

Plant‐originated glycoprotein (24 kDa) has an inhibitory effect on proliferation of BNL CL.2 cells in response to di(2‐ethylhexyl)phthalate

Di(2-ethylhexyl)phthalate (DEHP) is one of the many environmental chemicals that are widely used in polyvinyl chloride products, vinyl flooring, food packaging and infant toys. They cause cell proliferation or dysfunction of human liver. The purpose of this study is to investigate the inhibitory effect of a glycoprotein (24 kDa) isolated from Zanthoxylum piperitum DC (ZPDC) on proliferation of liver cell in the DEHP-induced BNL CL. 2 cells. [³H]-thymidine incorporation, intracellular reactive oxygen species (ROS), intracellular Ca²⁺ mobilization and activity of protein kinase C (PKC) were measured using radioactivity and fluorescence method respectively. The expression of mitogen-activated protein kinases [extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK)], activator protein (AP)-1 (c-Jun and c-Fos), proliferating cell nuclear antigen (PCNA) and cell cycle-related factors (cyclin D1/cyclin-dependent kinase [CDK] 4) were evaluated using Western blotting or electrophoretic mobility shift assay. The results in this study showed that the levels of [³H]-thymidine incorporation, intracellular ROS, intracellular Ca²⁺ mobilization and activity of PKCα were inhibited by ZPDC glycoprotein (100 µg/ml) in the DEHP-induced BNL CL. 2 cells. Also, activities of ERK, JNK and AP-1 were reduced by ZPDC glycoprotein (100 µg/ml). With regard to cell proliferation, activities of PCNA and cyclin D1/CDK4 were significantly suppressed at treatment with ZPDC glycoprotein (100 µg/ml) in the presence of DEHP. Taken together, these findings suggest that ZPDC glycoprotein significantly normalized activities of PCNA and cyclin D1/CDK4, which relate to cell proliferation factors. Thus, ZPDC glycoprotein appears to be one of the compounds derived from natural products that are able to inhibit cell proliferation in the phthalate-induced BNL CL. 2 cells.

Inhibitory effect of phytoglycoprotein (24 kDa) on hepatocarcinogenesis in N-nitrosodiethylamine-treated ICR mice

Hepatocellular carcinoma (HCC) is becoming one of the most prominent types of cancer in the world. For a long time in Korea Zanthoxylum piperitum DC (ZPDC) has been used in folk medicine to cure several cancers and inflammation. This study was designed to investigate whether ZPDC glycoprotein protected liver tissues against hepatocarcinogenic compounds such as N-nitrosodiethylamine (DEN). To study the chemopreventive effect of ZPDC glycoprotein on hepatocarcinogenesis, ICR mice were injected intraperitoneally with DEN (50 mg/kg) for four weeks. We evaluated the indicators of liver tissue damage (the activity of lactate dehydrogenase (LDH) and alanine aminotransferase (ALT), and thiobarbituric acid-reactive substances (TBARS)), antioxidative enzymes (activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx)), hepatocarcinogenic indicator (heat shock protein (HSP) 70) and hepatocarcinogenic signals (activity of nuclear factor (NF)-κB, cyclooxygenase (COX)-2, and matrix metalloproteinase (MMP)-9) using biochemical methods and immunoblot analysis. The results obtained from this study revealed that ZPDC glycoprotein (20 mg/kg) decreased the levels of LDH, ALT, and TBARS, whereas the activity of SOD and GPx increased in the DEN-treated ICR mice. With respect to the hepatocarcinogenic indicator and hepatocarcinogenic signals, HSP70, NF-κB, COX-2, and MMP-9 activity decreased. The findings suggested that ZPDC glycoprotein prevented damage to liver tissue caused by DEN in the experimental mouse model.