草连生物碱成分的研究

目的研究草连的生物碱成分。方法采用硅胶柱色谱法,进行单体化合物的分离纯化,通过波谱学方法鉴定其结构。结果分离并鉴定出7个原小檗碱型生物碱,分别为小檗碱、药根碱、巴马汀、非洲防己碱、黄连碱、甲基黄连碱和groen-landicine。结论7种生物碱均首次从草连中分得。草连所含生物碱的种类与黄连属其他植物中的生物碱种类不完全相同,生物碱种类可作为黄连属植物分类及生药鉴定的依据之一。     

黄连中原小檗型生物碱的分离与鉴定

目的:研究黄连中的原小檗型生物碱成分。方法:采用溶剂提取、色谱分离等手段对样品进行分离、纯化,根据波谱数据进行化合物结构鉴定。结果:从黄连中分离出6个生物碱成分,分别鉴定为盐酸小檗碱、盐酸黄连碱、盐酸药根碱、盐酸非洲防己碱、盐酸表小檗碱、盐酸巴马汀。结论:该方法简单、可行,可为黄连药材及其制剂的质量标准提供对照品。本试验为首次采用直接分离纯化的方法从黄连中得到盐酸表小檗碱。     

Protoberberine alkaloids as antimalarials

The protoberberine alkaloids berberine (1), palmatine (2), jatrorrhizine (3), and several berberine derivatives (4-10) were tested for antimalarial activity in vitro against Plasmodium falciparum and in vivo against Plasmodium berghei. The berberine derivatives 4-10 were designed and synthesized to maximize structural diversity within a modest set of compounds. Palmatine (2) and jatrorrhizine (3) were isolated as their chlorides from Enantia chlorantha. None of the protoberberine alkaloids was active in vivo, although compounds 1, 2, 3, 5, and 6 exhibited a potency comparable to that of quinine in vitro.     

黄连有效成分及其组合物对荷糖小鼠胰岛素和胰高血糖素样肽1(GLP-1)分泌影响的初步研究

目的：探讨黄连有效成分及其组合物对荷糖小鼠胰岛素和胰高血糖素样肽1（GLP-1）分泌的影响。方法：制备黄连粉末模拟胃液溶出物并检测其中盐酸小檗碱（BER）、黄连碱（COP）、盐酸药根碱（JAT）、阿魏酸（FA）、盐酸巴马丁（BM）含量,结合BER治疗2型糖尿病（T2D）有效剂量确立其他成分剂量,组合物剂量为5种化合物剂量的1/10倍量。对照组灌服葡萄糖（2 g·kg^-1）,药物组灌服等剂量葡萄糖及不同药物。各组分别在灌服的前1 h及灌服1 h时眼眶静脉采血、灌服1.5 h时摘眼球采血,分离血清,采用酶联免疫吸附法（ELISA）检测血清中的GLP-1和胰岛素含量。结果：BER、COP、JAT、组合物及黄连粉末均能在1,1.5 h升高荷糖小鼠血清胰岛素浓度,与对照组比较,P<0.05;BER、组合物及黄连粉均能在1,1.5 h升高荷糖小鼠血清GLP-1浓度,即促进GLP-1分泌;药物组其他成分呈现一定升高GLP-1水平作用趋势,但统计学差异不显著。结论：黄连及其部分活性成分能有效刺激小鼠GLP-1及胰岛素分泌,且低剂量组合物作用活性高于高剂量单个活性成分活性,黄连刺激肠道GLP-1分泌作用是其治疗T2D机制之一。     

一测多评法测定黄连须中的5种生物碱

目的 采用一测多评法同时测定黄连须中5种生物碱(盐酸小檗碱、盐酸药根碱、盐酸表小檗碱、盐酸黄连碱和盐酸巴马汀)的含量.方法 以盐酸小檗碱为内参物,采用HPLC法测定其与盐酸药根碱、盐酸表小檗碱、盐酸黄连碱、盐酸巴马汀的相对校正因子,利用该相对校正因子计算其他4种生物碱的含量;同时利用外标法测定5种生物碱的含量,比较两种测定方法的差异,验证一测多评法的可行性和准确性.结果 在线性范围内,盐酸小檗碱与盐酸药根碱、盐酸表小檗碱、盐酸黄连碱、盐酸巴马汀的相对校正因子分别为1.128、1.008、1.070、1.025,在不同实验条件下,相对校正因子的重复性良好,5种生物碱成分含量的计算值与实测值间无显著性差异.结论 一测多评法同时测定黄连须中5种生物碱的含量是可行的、准确的.     

Quaternary alkaloids of tinospora species

The occurrence of quaternary alkaloids in TINOSPORA (and PARABAENA) species (Menispermaceae) has been studied. The main components were generally the protoberberine bases berberine and palmatine, with jatrorrhizine an occasional minor constituent, and the aporphine base magnoflorine. Choline was also often present. Only magnoflorine was detected in the PARABAENA material examined.     

Monoamine oxidase inhibitors from rhizoma of Coptis chinensis

Three protoberberine alkaloids jatrorrhizine, berberine and palmatine were isolated from the monoamine oxidase (MAO) inhibitory fraction of the methanol extract of Coptis chinensis rhizoma. Jatrorrhizine was shown to inhibit non-competitively both MAO-A and -B from rat brain mitochondria with the IC50 values of 4 and 62 microM, respectively. Berberine only competitively inhibited MAO-A with an IC50 values of 126 microM whereas palmatine exhibited, up to 200 microM, no inhibition on any type of the enzyme. The structure-activity relationship was briefly discussed.     

大鼠口服泻心汤及其配伍组方中原小檗碱类成分尿排泄动力学

目的研究大鼠灌服泻心汤及其配伍组方中3种原小檗碱类生物碱(小檗碱、巴马汀、黄连碱)尿排泄动力学的变化。方法取SD大鼠20只随机分成黄连、大黄黄连、黄芩黄连、泻心汤4组,采集灌胃给药前及给药后不同时间段尿液,用HPLC法测定尿药浓度,由尿排泄-时间数据计算尿排泄动力学参数,采用方差分析比较各组中3种原小檗碱类生物碱尿排泄动力学参数的差异。结果3种原小檗碱类生物碱排泄速率常数相近,排泄半衰期为9~16h。与黄连组比较,泻心汤组、大黄黄连组、黄芩黄连组中黄连碱尿排泄量占给药量比值减少,其中泻心汤组与之比较差异具有统计学意义(P<0.05)。结论3种原小檗碱类生物碱具有类似的尿排泄特征,大黄、黄芩和黄连配伍能减少黄连碱的尿排泄量。     

Intestinal absorption mechanisms of berberine,palmatine, jateorhizine,and coptisine: involvement of P-glycoprotein

1. The absorption and transport mechanisms of berberine, palmatine, jateorhizine, and coptisine were studied using a Caco-2 cells uptake and transport model, with the addition of cyclosporin A and verapamil as P-glycoprotein (P-gp) inhibitors and MK-571 as a multidrug resistance-associated protein 2 (MRP₂) inhibitor.

2. In the uptake experiment, berberine, palmatine, jateorhizine, and coptisine were all taken into Caco-2 cells, and their uptakes were increased in the presence of cyclosporin A or verapamil.

3. In the transport experiment, P_app (AP-BL) was between 0.1 and 1.0?×?10⁶ cm/sec for berberine, palmatine, jateorhizine, and coptisine and was lower than P_app (BL-AB). ER values were all >2. Cyclosporin A and verapamil both increased P_app (AP-BL) but decreased P_app (BL-AB) for berberine, palmatine, jateorhizine, and coptisine; ER values were decreased by >50%. MK-571 had no influence on the transmembrane transport of berberine, palmatine, jateorhizine, and coptisine.

4. At a concentration of 1–100 μM, berberine, palmatine, jateorhizine, and coptisine had no significant effects on the bidirection transport of Rho123.

5. Berberine, palmatine, jateorhizine, and coptisine were all P-gp substrates; and at the range of 1–100 μM, berberine, palmatine, jateorhizine, and coptisine had no inhibitory effects on P-gp.

     

高效液相色谱法测定黄连药材中小檗碱型生物碱的含量

目的:分离并测定黄连药材中小檗碱型生物碱的含量。方法:采用ODS柱(15 mm×6.0 mm),以乙腈-50 mmol·L~(-1)磷酸二氢钾溶液(磷酸调 pH=3.0)(50:50),内含 25 mmol·L~(-1)SDS为流动相,345nm检测。结果:在以上条件下,药根碱、表小檗碱、黄连碱、巴马亭和小檗碱5种生物碱可以完全分离,测定了8种不同黄连药材中4种生物碱的含量。结论:本法可用于黄连药材的研究,也可用于黄连等含小檗碱中药材及制剂的常规检验。     

Simultaneous quantification of three alkaloids of Coptidis Rhizoma in rat urine by high-performance liquid chromatography: application to pharmacokinetic study

A high-performance liquid chromatographic method with ultraviolet detection was established and validated for quantification of three alkaloids (coptisine, palmatine and berberine) in rat urine. Following a single-step liquid-liquid extraction, the analytes were separated on a reversed-phase C(18) column with water-formic acid-triethylamine-methanol as the mobile phase at a flow rate of 1 ml/min. The linear ranges of the calibration curves were 1.6-160 ng/ml for all three alkaloids. The lower limit of quantification was 1.6 ng/ml for all three alkaloids. The within-batch accuracy was 90.4-108.3% for coptisine, 88.6-107.8% for berberine and 88.4-110.1% for palmatine. The between-batch accuracy was 99.3-100.3% for coptisine, 94.3-100.6% for berberine and 93.7-100.0% for palmatine. The within-batch and between-batch precisions were 相似文献   

Interaction of six protoberberine alkaloids with human organic cation transporters 1, 2 and 3

1.?Organic cation transporters (OCTs) play an important role in drug safety and efficacy. Protoberberine alkaloids are ubiquitous organic cations or weak bases with remarkable biological actives. This study was to elucidate the potential interaction of alkaloids (coptisine, jatrorrhizine, epiberberine, berberrubine, palmatine and corydaline) with OCTs using Madin–Darby canine kidney (MDCK) cells stably expressing human OCT1, OCT2 and OCT3.

2.?All the tested alkaloids significantly inhibited the uptake of MPP⁺, a model OCT substrate, in MDCK-hOCTs cells with the IC₅₀ of 0.931–9.65?μM. Additionally, coptisine, jatrorrhizine and epiberberine were substrates of all the hOCTs with the K_m of 0.273–5.80?μM, whereas berberrubine was a substrate for hOCT1 and hOCT2, but not for hOCT3, the K_m values were 1.27 and 1.66?μM, respectively. The transport capacity of coptisine in MDCK cells expressing the variants of hOCT1-P341L or hOCT2-A270S was significantly higher than that in wild-type (WT) cells with the Cl_int (V_max/K_m) of 379?±?7.4 and 433?±?5.7?μl/mg protein/min, respectively.

3.?The above data indicate that the tested alkaloids are potent inhibitors, and coptisine, jatrorrhizine, epiberberine and berberrubine are substrates of hOCT1, hOCT2 and/or hOCT3 with high affinity. In addition, the variants (OCT1-P341L and OCT2-A270S) possess higher transport capacity to coptisine than WT hOCTs.     

Increased methylmercury toxicity related to obesity in diabetic KK‐Ay mice

We examined the toxic effects of methylmercury (MeHg) in KK‐Ay type 2 diabetic mice to clarify how metabolic changes associated with type 2 diabetes mellitus affect MeHg toxicity. MeHg (5 mg Hg kg ^–1day^–1 p.o.) was given to 4‐week‐old male KK‐Ay and C57BL/6J (BL/6) mice three times per week for 6 weeks. Average body weights (BW) of vehicle‐treated BL/6 and KK‐Ay mice were 16.3 and 16.4 g respectively on the first day, and 24.8 and 42.3 g respectively on the last day of the experiment. MeHg‐treated KK‐Ay mice began to lose weight about 5 weeks after MeHg administration. Six of seven MeHg‐treated KK‐Ay mice showed hind‐limb clasping in the final stage of the experiment. The mean blood mercury level of MeHg‐treated KK‐Ay mice reached a maximum of 9.8 µg ml^–1, whereas that of the MeHg‐treated BL/6 mice was 2.8 µg ml^–1 after 10 days of treatment. The average total mercury concentrations in the cerebrum and epididymal fat pad were 7.4 and 0.57 µg g^–1, respectively, for BL/6 mice and 27 and 1.6 µg g^–1, respectively, for KK‐Ay mice. In MeHg‐treated KK‐Ay mice with neurological symptoms, CD204‐positive macrophages were observed in the brain, kidney and spleen, indicating CD204 could be a marker for injured tissues. BW loss and significant pathological changes were not observed in other groups of mice. These results indicate that body fat gain in type 2 diabetes mellitus and low mercury accumulation in adipose tissue increased MeHg concentrations in organs and enhanced toxicity in KK‐Ay mice at the same dose of MeHg per BW. Copyright © 2013 John Wiley & Sons, Ltd.     

正交试验优化提取黄连中小檗碱型生物碱

目的:研究黄连中小檗碱型生物碱的最佳提取工艺。方法:采用正交试验设计,以小檗碱、巴马汀、黄连碱、表小檗碱和药根碱的总含量为考察指标,考察乙醇浓度、提取时间、提取次数、乙醇用量4个因素对提取结果的影响。结果:最佳提取工艺为8倍量的50%乙醇,回流提取4次(每次2h)。结论:本工艺重现性好,测定方法可行,为大批量黄连提取生物碱提供了理论依据。     

胶束色谱分析黄连及含黄连中成药中小檗碱型生物碱的研究

本文报道用胶束色谱法分离测定黄连(黄柏)及含黄连(黄柏)中成药中黄连碱、药根碱、巴马亭和小槃碱。考察了固定相、胶束浓度、有机改性剂浓度及pH对组分保留行为的影响。实验结果表明;影响该色谱分离的主要因素为固定相极性和胶束浓度。同时在肢束流动相中加入甲醇改性剂,可在室温下显著地提高柱效。用键合苯基固定相,以3.5%十二烷基硫酸钠/0.1 N酒石酸—甲醇(30:70)胶束溶液作流动相,测定了十几种黄连(黄柏)及含黄连(黄柏)的中成药。     

Determination of the alkaloid content in different parts of some Mahonia plants by HPCE

The contents of three quaternary alkaloids (berberine, palmatine, jatrorrhizine) in different parts of the genus Mahonia were determined by high-performance capillary electrophoresis (HPCE). The background electrolyte system composed of 0.1 M phosphate buffer (pH 7.0)-methanol (2:1 V/V) was found to be the most suitable solution for this separation. Brucine was used as internal standard. The linear calibration ranges were 0.004986-0.4986 mg ml-1 (r = 0.9990, n = 5) for berberine, 0.005049-0.5049 mg ml-1 (r = 0.9996, n = 5) for palmatine, and 0.005058-0.5058 mg ml-1 (r = 0.9984, n = 5) for jatrorrhizine. The relative standard deviations were 1.56%, 1.02%, and 1.60% for berberine, palmatine, and jatrorrhizine (n = 6), respectively. The recoveries were determined to be 96.00-101.66% for berberine, 100.15-102.97% for palmatine, and 96.68-102.44% for jatrorrhizine. By using proposed HPCE method, three alkaloids were well-separated within only 5.0 min.     

用LC／MS／MS和RP—HPLC法对泻心汤煎煮产生的沉淀物的研究

目的：用ＬＣ／ＭＳ／ＭＳ和ＲＰ－ＨＰＬＣ法研究沔心汤煎煮产生的沉淀物。方法：利用ＬＣ－ＭＳ－ＭＳ法对淀心肠沉淀物进行了初步研究,又利用ＲＰ－ＨＰＬＣ法,测定了淀心汤沉淀物中小檗碱在小鼠体内的吸收。结果：发现其中含有小檗碱、巴马丁碱、黄连碱、药根碱,但在给小鼠灌服淀心汤沉淀物后２４ｈ内的几个时间段中,均未能从血浆中测到小檗碱。结论：淀心汤沉淀物进入体内后小檗碱并未进入血液,推测如果淀心汤沉淀物具有恬     

Application of <Superscript>1</Superscript>H-NMR spectroscopy to validation of berberine alkaloid reagents and to chemical evaluation of Coptidis Rhizoma

Berberine, palmatine, and coptisine are major pharmacologically active protoberberine alkaloids in Coptidis Rhizoma, and have been used as indices for chemical evaluation of the crude drug. ¹H-NMR spectroscopy was applied to determination of purities of commercial reagents of protoberberine alkaloids. The purities of the alkaloids were calculated from the ratios of the intensities of the H-13 singlet signal at about δ 8.7 ppm of target protoberberine alkaloids to integration of a hexamethyldisilane (HMD) signal at 0 ppm. The concentration of HMD was corrected with SI traceability using potassium hydrogen phthalate of certified reference material (CRM) grade. The purity of the reagent estimated by the ¹H-NMR was, in general, lower than that claimed by the manufacturer, leading to over-estimation of the alkaloid contents of Coptidis Rhizoma when determined by HPLC. The present quantitative ¹H-NMR method was also applicable to direct determination of protoberberine alkaloid contents in Coptidis Rhizoma.     

黄连有效成分盐酸小檗碱、盐酸药根碱、黄连碱及其混合物对NCI-H716细胞分泌胰高血糖素样肽1(GLP-1)的影响

目的:研究黄连有效成分对NCI-H716细胞分泌胰高血糖素样肽1(GLP-1)的影响。方法:分别设置空白对照组、盐酸药根碱(JAT)组、盐酸小檗碱(BER)组、黄连碱(COP)组、盐酸巴马汀(BM)及BER十COP十JAT高剂量组和BER+COP+JAT低剂量组,同时以BER组为阳性对照组。采用CCK-8法检测各组药物不同浓度下对细胞增殖的抑制作用,确定药物浓度范围;采用酶联免疫吸附法(ELISA)检测NCI-H716细胞分泌GLP-1的量;采用聚合酶链式反应(PCR)检测NCI-H716细胞表达GLP-1RNA的量。结果:COP、JAT,以及高、低浓度BER+COP+JAT均能促进NCI-H716细胞分泌GLP-1(P<0.05),其中,BER+COP+JAT高浓度组及BER+COP+JAT低浓度组促GLP-1分泌作用均优于阳性对照组(P<0.05);COP、JAT组与阳性对照组相比无显著性差异(P>0.05)。结论:JAT、COP对NCI-H716细胞分泌GLP-1均有促进作用,且BER+COP+JAT混合组效果优于其单体组。