甘草抗动脉粥样硬化机制的网络药理学研究

利用网络药理学方法探讨甘草在抗动脉粥样硬化中的分子机制。本研究利用中医药系统药理学数据库和分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)分析甘草中的有效活性成分,并获得有效成分的作用靶点。通过Cytoscape软件构建可视化靶点互相作用网络,对网络中的关键靶点进行基因本体(GO)富集分析和KEGG通路富集分析。结果显示甘草中40种有效活性成分的预测靶点共97个,47个靶点与动脉粥样硬化(AS)相关,其中18个是血管保护药物和脂质修饰药物的作用靶点,表明甘草可作为调控AS发展的药物。基于97个预测靶点的GO富集分析,发现甘草可参与多种生物学过程,尤其是应对外源性刺激,以及参与细胞凋亡等过程。通过构建甘草靶点与AS疾病靶点相互作用网络(PPI),确定了AKT1、MAPK3、MAPK1、JUN和CASP3等关键靶点,并对关键靶点进行KEGG富集分析,结果表明甘草主要影响调控细胞增殖、生存以及凋亡的细胞信号转导相关通路,并激活先天免疫相关信号通路,调节炎性细胞因子释放,从而发挥抗动脉粥样硬化作用。甘草具有多成分、多靶点、多途径的作用特点,主要通过PI3K-AKT信号途径、MAPK信号途径、NOD样受体信号通路调控细胞增殖和凋亡,同时发挥免疫调控作用,从而影响动脉粥样硬化的发展,由此可见,甘草可作为动脉粥样硬化疾病治疗的候选中草药。     

液相色谱/离子阱质谱法研究何首乌中糖苷类化合物

采用液相色谱／离子阱质谱（HPLC／IT-MS）联用技术研究何首乌中糖苷类化合物,并对大黄素甲醚糖苷和大黄素糖苷的分子离子或准分子离子的离子化机理进行探讨。实验采用反相C18色谱柱,二元线性梯度洗脱,分离出12个主要成分。利用质谱的诱导碰撞解离技术获得碎片裂解信息,结合文献鉴定出9种糖苷的化学结构,其中顺式-2,3,5,4＇-四羟基二苯乙烯-2-O-β-D-毗喃葡萄糖苷、决明酮-8—O-（6＇-O-乙酰基）-β—D-吡喃葡萄糖苷和大黄素甲醚-8-O-（6＇-O-乙酰基）-β-D-吡喃葡萄糖苷为首次鉴定。     

Occurrence and fate of fetal lumbar rib induced by Scutellariae radix in rats

BACKGROUND: This study was conducted to evaluate the occurrence and fate of fetal lumbar rib induced by Scutellariae radix (SR) in rats. METHODS: Water extracts of SR were orally administered to pregnant rats from day 7 to day 17 of gestation at a dose of 186 mg/kg/day, equivalent to 25 g/kg of starting material, representing a 100‐fold increase over typical human intake level. RESULTS: The incidence of fetal lumbar rib in the SR‐treated group was increased on gestational day 20 and then decreased on postnatal day 50. The weight of fetuses in the SR‐treated group tended to be less than that in the control group. Alkaline phosphatase in SR‐treated dams was increased on gestational day 20, but was decreased on postnatal day 50. There were no significant differences between the vehicle control and SR‐treated groups in maternal body weight, embryological, histopathological, hematological, and serum biochemical changes. CONCLUSIONS: The present data suggest that the appearance of lumbar rib induced by SR is a transient fetal variation rather than teratogenicity or maternal toxicity. Birth Defects Res (Part B) 89:201–206, 2010. © 2010 Wiley‐Liss, Inc.     

Protective Effects of Platycodi radix on Alcohol-Induced Fatty Liver

The protective effects of Platycodi radix (PR), the root of Platycodon grandiflorum A. DC, on alcohol-induced fatty liver and possible mechanisms involved in this protection were investigated in rats. Administration of PR significantly prevented alcohol-induced elevation of serum and liver lipids. Furthermore, PR treatment normalized hepatic liver fatty acid binding protein (L-FABP) expression and cytochrome P450 2E1 (CYP2E1) activity in alcohol-treated rats. These results suggest that inhibition of CYP2E1 and regulation of L-FABP by PR play an important role in alcohol-induced hepatoprotection.     

A comprehensive review on the stinging nettle effect and efficacy profiles. Part II: Urticae radix

Nettle root is recommended for complaints associated with benign prostatic hyperplasia (BPH). We therefore conducted a comprehensive review of the literature to summarise the pharmacological and clinical effects of this plant material. Only a few components of the active principle have been identified and the mechanism of action is still unclear. It seems likely that sex hormone binding globulin (SHBG), aromatase, epidermal growth factor and prostate steroid membrane receptors are involved in the anti-prostatic effect, but less likely that 5alpha-reductase or androgen receptors are involved. Extract and a polysaccharide fraction were shown to exert anti-inflammatory activity. A proprietary methanolic nettle root extract and particular fractions inhibited cell proliferation. Isolated lectins (UDA) were shown to be promising immunomodulatory agents, having also anti-viral and fungistatic effects. However, despite these in vitro studies it is unclear whether the in-vitro or animal data are a surrogate for clinical effects. The clinical evidence of effectiveness for nettle root in the treatment of BPH is based on many open studies. A small number of randomised controlled studies indicate that a proprietary methanolic extract is effective in improving BPH complaints. However, the significance and magnitude of the effect remains to be established in further confirmatory studies before nettle root treatment may be accepted in the guidelines for BPH treatment. The risk for adverse events during nettle root treatment is very low, as is its toxicity. Pre-clinical safety data remain to be completed.     

DNA barcoding Chinese medicinal Bupleurum

We tested 4 markers, namely nuclear internal transcribed spacer 2 (ITS2), psbA-trnH, matK, and rbcL, to evaluate these candidate DNA barcodes for distinguishing Bupleuri radix (Chaihu) from its adulterants. 51 plant samples of Bupleurum representing 19 species were collected from different areas in China. Amplification and sequencing were attempted for all the 4 candidate barcode regions, whose validity was assessed in terms of the success rate of PCR amplification and sequencing, differential intra- and inter-specific divergences, DNA barcoding gap and the ability to discriminate species. The results showed that ITS2 had the best performance in identifying Bupleurum with an identification efficiency of 73.68%, which, after combining with psbA-trnH, increased to 83.33%. We further evaluated the efficiency of ITS2 for discriminating the species of Bupleurum using a large database from GenBank, which archived data of 223 samples from 74 species, and ITS2 successfully discriminated 64.13% of the samples at the species level. In conclusion, the ITS2 can serve as a potentially useful barcode for Bupleurum species, with psbA-trnH as a supplementary locus.     

当归对实验性血虚便秘模型小鼠结肠组织形态和黏液分泌的影响

目的：观察当归对实验性血虚便秘模型小鼠的治疗作用及其对小鼠结肠组织形态和黏液分泌的影响。方法：60只KM小鼠,雌雄各半,随机分为6组（n=10）：正常对照组、血虚便秘组、阳性对照组、当归高、中、低剂量组。除正常对照组小鼠外,其余各组小鼠灌胃给予复方地芬诺酯（DNP）,皮下注射乙酰苯肼（APH）,腹腔内注射环磷酰胺（CPA）,复制血虚便秘小鼠模型。从实验的第14天起,当归组小鼠灌胃给予不同剂量的当归煎液（16.67,8.33,4.17 g/kg）,阳性对照组给予常通舒颗粒（5 g/kg）,血虚便秘模型组和正常对照组小鼠给予等体积生理盐水,每日1次,连续28 d。观察小鼠一般状态,检测首粒黑便排出时间（FBDT）、粪便和结肠含水量,取结肠组织进行HE和AB-PAS染色,观察小鼠结肠组织形态变化和黏液分泌情况。结果：与正常对照组比较,血虚便秘组小鼠出现血虚和便秘体征,FBDT显著延长、粪便和结肠含水量显著降低（P<0.01）,结肠黏膜和大肠腺萎缩、黏膜层变薄（P<0.01）,黏液分泌减少。与血虚便秘模型组比较,3个剂量的当归均能改善血虚便秘体征,显著缩短FBDT（P<0.05,P<0.01）,显著升高小鼠结肠含水量（P<0.05,P<0.01）;高、中剂量的当归能显著升高小鼠粪便含水量（P<0.05）;3个剂量当归均能改善血虚便秘模型小鼠的结肠黏膜和大肠腺萎缩,增加结肠黏液分泌,改善结肠润滑功能。结论：当归可改善结肠黏膜萎缩、增加结肠黏液分泌,从而软化粪便促进粪便排出。     

Hwangryun-Hae-Dok-tang (Huanglian-Jie-Du-Tang) extract and its constituents reduce ischemia-reperfusion brain injury and neutrophil infiltration in rats

The preventive effect of Hwangryun-Hae-Dok-tang (HHDT, Huanglian-Jie-Du-Tang), a Chinese herbal medicine, and its ingredients on ischemia/reperfusion-induced brain injury was evaluated in the rat brain. HHDT consists of four herbs, namely, Coptidis rhizoma, Scutellariae radix, Phellodendri cortex, and Gardeniae fructus. Ischemia was induced by intraluminal occlusion of the right middle cerebral artery for 120 min and reperfusion was continued for 22 h. HHDT (200 mg/kg), Coptidis rhizoma (100 mg/kg), Scutellariae radix (100 mg/kg), Phellodendri cortex (100 mg/kg), and Gardeniae fructus (100 mg/kg) were orally administered, promptly prior to reperfusion and 2 h after reperfusion. Baicalein, a component of Scutellariae radix, was also examined at a dosage of 50 mg/kg given 2 h apart, promptly prior to and 2 h after reperfusion. Total infarction volume in the ipsilateral hemisphere of ischemia/reperfusion rats was significantly lowered by treatment with HHDT, Scutellariae radix, and balicalein. However, the other ingredient of HHDT did not show any ameliorating effects on total infarction volume. The inhibiting effect of Scutellariae radix on total infarction volume was much higher than that of the others. In addition, HHDT, Scutellariae radix, and baicalein significantly inhibited myeloperoxidase (MPO) activity, an index of neutrophil infiltration in ischemic brain tissue at about the same rate (30%). There was marked mismatch between total infarction volume and MPO activity in the Scutellariae radix-treated rats but not in the HHDT- and baicalein-treated groups. Our findings suggest that Scutellariae radix as an ingredient of HHDT plays a crucial protective role in ischemia-induced brain injury. In addition, it is apparent that the effect of Scutellariae radix is the result, in part, of baicalein, a compound contained in Scutellariae radix.     

A novel homodimeric lectin from Astragalus mongholicus with antifungal activity

A novel lectin (AMML) was isolated from a Chinese herb, i.e., the roots of Astragalus mongholicus, using a combination of ammonium sulfate fraction and ion exchange chromatographies. The molecular mass of intact AMML was determined to be 66,396 Da by MALDI-TOF mass spectrometry and 61.8 kDa by gel filtration, respectively. AMML was a dimeric protein composed of two identical subunits each with a molecular mass of 29.6 kDa. The lectin was a glycoprotein with a neutral carbohydrate content of 19.6%. The purified lectin hemagglutinated both rabbit and human erythrocytes, and showed preference for blood types O (native) and AB (trypsin-treated). Among various carbohydrates tested, the lectin was best inhibited by D-galactose and its derivatives with pronounced preference for lactose (3.13 mM). N-terminal amino acid sequence of AMML was determined as ESGINLQGDATLANN. The optimal pH range for lectin activity was between pH 4.5 and 7.5, and the lectin was active up to 65 degrees C. It also exerted antifungal activity against Botrytis cincerea, Fusarium oxysporum, Colletorichum sp., and Drechslera turcia but not against Rhizoctonia solani and Mycosphaerella arachidicola.     

甘草根中甘草酸的免疫组织化学定位研究

应用免疫组织化学定位和HPLC分析方法,对人工种植乌拉尔甘草根中的甘草酸分布和积累变化规律进行分析,以阐明药用甘草根在发育过程中甘草酸的积累变化规律,探讨甘草药材质量形成的内在机制。结果表明:(1)甘草酸主要分布在根的次生韧皮部薄壁细胞和维管射线细胞中,且主要积累在这些细胞的细胞壁上。(2)HPLC分析显示,主根的韧皮部中甘草酸含量最高,其次是木质部,根皮中含量最少。(3)甘草酸在主根中的含量随着生长年限的增加而提高,2、3年生甘草根中甘草酸含量上升较快,3年后甘草酸的含量达到3.66%,超过了国家药典规定的甘草酸含量标准。