Zanthoxylum avicennae extract enhances GSK‐3β to attenuate β‐catenin via phosphatase 2A to block metastatic effects of HA22T cells and hepatocellular carcinoma xenografted nude mice

Hepatocellular carcinoma (HCC) metastasis is often associated with the activation of Wnt/β‐catenin signaling pathway. Zanthoxylum avicennae (Ying Bu Bo, YBB), a traditional herb with hepatoprotective effect, has been proven to inhibit human HCC in in vivo models however, the in vitro and in vivo effect of YBB on tumor metastasis is not clear yet. To determine whether YBB could inhibit HA22T human HCC cell by acting on β‐catenin metastatic signaling in vitro and in vivo, HA22T cells were treated with different concentrations of YBB extracts (YBBE) and analyzed by Immunofluorescence staining assay, western blot analysis, siRNA mediated gene knock‐down assays and co‐immunoprecipitation assay. Additionally, the HA22T‐implanted xenograft nude mice were used to confirm the assessed cellular effects. Mice treated with YBBEs showed a strong increasing trend in PP2Acα, GSK‐3β, APC, and β‐TrCP/HOS levels, however the expression of β‐catenin, p‐GSK‐3β, TBX 3, and IL8 proteins showed a decreasing trend. YBBE significantly downregulated the nuclear and cytosolic β‐catenin levels by facilitating the proteosomal degradation of β‐catenin. Moreover, as observed by co‐immunoprecipitation assay, YBBE directly promoted the protein interactions between GSK‐3β, β‐TrCP, APC, PP2A, and β‐catenin. In conclusion, both in vitro and in vivo models clearly demonstrated that YBBE inhibits β‐catenin involved metastatic signaling in highly metastatic HA22T cells through PP2A activation.     

椒目油改善铁过载雌性去势大鼠骨质疏松的作用研究

目的 椒目油（Zanthoxylum bungeanum Maxim seed oil, ZBSO）对铁过载雌性去势SD大鼠骨质疏松的保护作用及机制初探。方法 将40只雌性SD大鼠随机分为假手术组（SHAM）、去势组（OVX）、铁过载组（Fe）、铁过载+椒目油组（Fe+ZBSO）。除SHAM组外，其余各组均实施卵巢切除术，Fe、Fe+ZBSO组以100 mg/kg·bw/w腹腔注射右旋糖酐铁，此外Fe+ZBSO组以3.46 ml/kg·bw/d干预12周。干预结束后，取股骨Micro-CT扫描骨结构和骨密度；ELISA法测定血清Ⅰ型胶原交联羧基端肽（CTX Ⅰ）、Ⅰ型前胶原氨基端原肽（PⅠNP）、8-羟基脱氧鸟苷（8-OHdG）、3-硝基酪氨酸（3-NT）的含量；比色法测定血清丙二醛（MDA）的浓度。结果 Fe、Fe+ZBSO组相较于OVX组，血清铁蛋白含量上升（q=19.230,q=10.050,P＜0.001）；相较于SHAM组，OVX（Z=3.977,P＜0.001）、Fe（Z=4.542,P＜0.001）、Fe+ZBSO（Z=3.241,P=0.004）组子宫脏器指数降低。相较于Fe组，Fe+ZBSO组股骨结构更完整，骨密度上升（t=6.222,P＜0.001）；骨代谢结果显示，Fe+ZBSO组CTX Ⅰ降低（t=2.603,P=0.019）、PⅠNP升高（t=2.839,P=0.012）；氧化损伤标志物结果显示，Fe+ZBSO组3-NT降低（t=2.894,P=0.015）、MDA降低（t=4.456,P＜0.001），8-OHdG无差异（F=0.101,P=0.959）。结论 椒目油能改善铁过载去势雌性SD大鼠的股骨骨骺端结构，提高骨密度，调节骨代谢，这可能是通过降低氧化损伤实现的。     

HPLC同时测定甘肃产竹叶椒根中白藓碱和茵芋碱含量

目的建立同时测定甘肃产竹叶椒根中白藓碱和茵芋碱含量的高效液相色谱方法。方法采用Kromasil-C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(65∶35),检测波长254 nm,流速1.0 m L/min,柱温30℃。结果白藓碱进样量在0.205~2.05μg(r=0.999 9)、茵芋碱进样量在0.199 6~1.996μg(r=0.999 7)范围内线性关系良好,白藓碱和茵芋碱的平均加样回收率分别为99.69%(RSD=0.57%)、99.49%(RSD=1.04%)。结论该方法简便、准确,适用于甘肃产竹叶椒根中白藓碱和茵芋碱的含量测定。     

花椒挥发油对离体家兔结肠平滑肌收缩功能的作用

目的研究花椒挥发油（essential oil of Zanthoxyhm Maxim,EOZM）对离体家兔结肠平滑肌的作用及其可能机制。方法分别观察EOZM和维拉帕米（verapamil,Ver）对结肠自律性收缩的影响、对乙酰胆碱（acetylcholine,ACh）和氯化钙（calciumchloride,CaCI2）引起的结肠收缩的影响、对ACh诱导的依内钙性收缩和依外钙性收缩的影响。结果EOZM和Ver均可抑制结肠自律性收缩以及ACh和CaCI2引起的结肠收缩,且具有剂量依赖性;EOZM对ACh诱导的依内钙性收缩和依外钙性收缩均有抑制作用;Ver仅对ACh诱导的依内钙性收缩有抑制作用,对ACh诱导的依外钙性收缩则没有影响。结论EOZM可能主要通过阻断钙通道,减少外钙内流和内钙释放从而抑制家兔结肠平滑肌的收缩。     

Effect of Zanthoxylum schinifolium on TNF-α-induced vascular inflammation in human umbilical vein endothelial cells

Pro-inflammatory cytokines induce the injury of endothelial cells in response to increases of adhesion molecules, leading to vascular inflammation and the development of atherosclerosis. In this study, we evaluated an ethanol extract of Zanthoxylum schinifolium (EZS) to determine if it inhibits the expressions of cellular adhesion molecules in human umbilical vein endothelial cells (HUVEC). When pretreatment of HUVEC with EZS, EZS suppressed the expression levels of cell adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and E-selectin induced by TNF-α. The adhesion of HL-60 cells to TNF-α-induced endothelial cells was decreased significantly in a concentration-dependent manner. Furthermore, TNF-α-induced MCP-1 and IL-8 mRNA expression levels were also attenuated by pretreatment with EZS. In addition, EZS suppressed TNF-α-induced production of reactive oxygen species (ROS). EZS inhibited NF-κB activation and IκB-α phosphorylation induced by TNF-α, subsequent degradation of IκB-α. Finally, EZS inhibited TNF-α-induced p38 MAPK and c-Jun N-terminal kinase (JNK) phosphorylation. Taken together, these results demonstrate that EZS suppresses vascular inflammatory process, which may be closely related to the inhibition of ROS, JNK, p38 MAPK and NF-κB activation in HUVEC.     

Effects of canthin-6-one alkaloids from Zanthoxylum chiloperone on Trypanosoma cruzi-infected mice

Canthin-6-one (1), isolated from Zanthoxylum chiloperone (Rutaceae), possesses a broad sprectum of antifungal and leishmanicidal activities. In this study, we have examined the antiparasitic effects of canthin-6-one (1), 5-methoxycanthin-6-one (2), canthin-6-one N-oxide (3), as well as that of the total alkaloids of Zanthoxylum chiloperone stem bark, in Balb/c mice infected either acutely or chronically with Trypanosoma cruzi. The compounds were administered orally or subcutaneously at 5mg/kg/day for 2 weeks, whereas the alkaloidal extract was given at 50mg/kg/day for 2 weeks. The antiparasitic activity was compared with that of benznidazole given at 50mg/kg/day for 2 weeks. In the case of acute infection, parasiteamia was significantly reduced following oral treatment with canthin-6-one (1). Moreover, the total alkaloids of Zanthoxylum chiloperone stem bark led to high levels of parasitological clearance. Seventy days post-infection, the serological response in the acute model was significantly different between oral canthin-6-one (1) and benznidazole-treated mice. Chronic model of the disease showed that both canthin-6-one (1) and the alkaloidal extract at the above dosage induced 80-100% animal survival compared to untreated controls. These results indicate that canthin-6-one (1) exhibits trypanocidal activity in vivo in the mouse model of acute or chronic infection. This is the first demonstration of anti-Trypanosoma cruzi activity for a member of this chemical group (canthinones). Considering the very low toxicity of canthin-6-one (1), our results suggest that long-term oral treatment with this natural product could prove advantageous compared to the current chemotherapy of Chagas disease.     

Protective effects of leaf extract of Zanthoxylum ailanthoides on oxidation of low-density lipoprotein and accumulation of lipid in differentiated THP-1 cells

It has been reported that extracts of stem and leaf of Zanthoxylum ailanthoides (ZLE) possess antioxidative properties. However, the biological importance of the ZLE is not well known. In our preliminary study, it showed that ZLE prepared from 75% alcohol highly contains flavonoids (5.8%). By HPLC analysis, it shows that the ZLE consists of flavonoid glycosides including rutin and hyperoside. We investigate the effects of ZLE on the oxidation of low-density lipoprotein (LDL; d = 1.019–1.063 g/mL) and the uptake of lipid in macrophage. Firstly, we explored the effect of ZLE on the oxidation of LDL by employing copper (II) sulfate (CuSO₄) as an oxidative inducer. Oxidation was monitored by the formation of conjugated diene and thiobarbituric acid relative substances (TBARS), relative electrophoretic mobility (REM), and fragmentation of apolipoprotein B-100 (Apo B). Our data showed that ZLE reduced the oxidation properties of LDL induced by CuSO₄. In addition, ZLE inhibited lipid accumulation in differentiated THP-1 cells treated with ox-LDL involving decreasing the expression of scavenger receptors such as scavenger receptor class AI (SR-AI) and CD36, which belongs to the class B scavenger receptor (SR-B). These results demonstrate the protective effect of ZLE on LDL oxidation and lipid accumulation in macrophage.     

Comparative repellency of 38 essential oils against mosquito bites

The mosquito repellent activity of 38 essential oils from plants at three concentrations was screened against the mosquito Aedes aegypti under laboratory conditions using human subjects. On a volunteer's forearm, 0.1 mL of oil was applied per 30 cm2 of exposed skin. When the tested oils were applied at a 10% or 50% concentration, none of them prevented mosquito bites for as long as 2 h, but the undiluted oils of Cymbopogon nardus (citronella), Pogostemon cablin (patchuli), Syzygium aromaticum (clove) and Zanthoxylum limonella (Thai name: makaen) were the most effective and provided 2 h of complete repellency.From these initial results, three concentrations (10%, 50% and undiluted) of citronella, patchouli, clove and makaen were selected for repellency tests against Culex quinquefasciatus and Anopheles dirus. As expected, the undiluted oil showed the highest protection in each case. Clove oil gave the longest duration of 100% repellency (2-4 h) against all three species of mosquito.     

刺异叶花椒根中香豆素类成分

目的:对刺异叶花椒Zanthoxylum dimorphophyllum var. spinifolum根的化学成分进行研究.方法:采用现代色谱分离技术进行化学成分的分离,运用波谱方法确定结构.结果:从中分离得到5个化合物,分别鉴定为5-甲氧基-7-羟基-6-(3'-甲基-2',3'-二羟基丁基)-香豆素(Ⅰ),6-(3'-甲基-2',3'-二羟基丁基)-7-甲氧基-8-(3″-甲基-丁-2″-烯基)-香豆素(Ⅱ),6-(3'-甲基-2',3'-二羟基丁基)-7-羟基-8-(3″-甲基-丁-2″-烯基)-香豆素(Ⅲ),6-(3'-甲基-2',3'-环氧丁基)-7-甲氧基-8-(3″-甲基-丁-2″-烯基)-香豆素(Ⅳ),7-甲氧基-8-(3'-甲基-丁-2'-烯基)-香豆素(Ⅴ).结论:以上化合物均为首次从该植物中分得.     

花椒及其同类品的形态组织学研究

据文献记载和作者实际调查,我国各地花椒属(Zanthoxylum L.)植物果实作药用者计约18种。本文先报道其中8种果皮的形态组织学研究结果:花椒Zanthoxylum bungeanum Maxim.青椒Z.schinifolium Sieb.et Zucc.,竹叶椒Z.armatum DC.,野花椒Z.simulans Hance,勒(木党)Z.avicennae(Lam.)DC.,椿叶花椒Z.ailanthoides Sieb.et Zucc.,朵椒Z.molle Rehd.及两面针Z.nitidum(Roxb.)DC.。本文附形态组织图并分别列有生药性状及显微特征检索表。