花旗松素对β-淀粉样肽损伤神经元的保护作用及机制

目的: 研究花旗松素对 β-淀粉样肽(β-amyloid peptide,Aβ)损伤神经元的保护作用及其机制. 方法: 从新生乳鼠大脑皮层中分离纯化神经元,与Aβ(5 μmol·L^-1)、不同浓度花旗松素(20~100 μmol·L^-1)共同孵育24 h,CCK8法检测细胞存活率,Hoechst 33258染色和Annexin V-FITC/PI检测细胞凋亡,分光光度法检测半胱天冬酶-3(caspase-3)活性的变化. 结果: 在细胞活力实验中,与正常组相比,模型组细胞活力显著下降(49.2±1.3) %,不同浓度的花旗松素可以提高神经元活力,其中80 μmol·L^-1花旗松素给药组细胞活力增至(68.7±3.2) %,与模型组相比有极显著差异;细胞凋亡结果显示,与正常组相比较,模型组细胞凋亡率为(50.8±1.5) %(P<0.01),不同浓度的花旗松素给药组可以降低细胞凋亡率,其中40 μmol·L^-1 花旗松素凋亡率为(41.5±2.7) %,与模型组比较呈显著性差异(P<0.05);凋亡酶caspase-3的活性与正常组(0.12±0.02) U·μg^-1比较,模型组的caspase-3的活性升高(2.37±0.16) U·μg^-1,P<0.01),与模型组相比,40 μmol·L^-1花旗松素组caspase-3显著降低(1.77±0.07) U·μg^-1, P<0.01).花旗松素能明显提高Aβ损伤神经元的细胞活力,抑制神经元的凋亡,降低凋亡酶caspase-3的活性. 结论: 花旗松素对Aβ损伤神经元具有保护作用,其机制可能与抑制caspase-3活性从而实现抗凋亡作用有关.     

五子承气汤对大鼠高尿酸血症的影响

目的: 研究五子承气汤对高尿酸血症大鼠的影响. 方法: 将雄性SD大鼠60只随机分为6组,即:正常组、模型组、五子承气汤高、中、低剂量组(104,52,26 g·kg^-1)、阳性对照苯溴马隆(10 mg·kg^-1)组,采用腺嘌呤(100 mg·kg^-1,ig,qd × 28 d)和氧嗪酸钾(250 mg·kg^-1,ip,1次/周,连续4周)制备高尿酸血症大鼠模型.造模4周后,检测各组大鼠尿尿酸(UA),血尿酸(UA),血黄嘌呤氧化酶(XOD),血肌酐(SCr),尿素氮(BUN). 结果: 与正常组相比,模型组大鼠的尿UA显著降低,血UA,XOD,Cre及BUN 明显升高,差异具有显著性(P<0.01);五子承气汤低、中、高剂量组尿UA分别为(146.3±22.5),(202.2±53.8),(218.1±62.6) μmol·L^-1,显著高于模型组(89.8±27.1)μmol·L^-1(P<0.01,P<0.05);血UA分别为(207.3±42.5),(177.1±50.5),(158.0±49.0)μmol·L^-1,显著低于模型组(285.0±54.1)μmol·L^-1(P<0.01,P<0.05);五子承气汤还显著降低高尿酸血症大鼠血清XOD活性,显著降低SCr,BUN含量( P<0.01,P<0.05). 结论: 五子承气汤明显促进尿UA排泄,降低血UA含量,抑制血清XOD活性,改善肾功能,对大鼠高尿酸血症有一定的防治作用.     

侧柏酮对人γδT细胞功能的影响

目的: 探讨侧柏酮对人γδ T细胞功能的影响. 方法: 异戊烯焦磷酸/isopentenyl pyrosphate(IPP)法体外扩增人外周血γδ T细胞.各浓度侧柏酮作用人γδ T细胞48 h,CCK-8法检测其增殖力,流式细胞术测其表型、细胞周期、细胞凋亡及表达CD107a,穿孔素(perforin),粒酶B(granzyme B)的情况. 结果: 300～600 μmol·L^-1侧柏酮明显抑制人γδ T细胞生长(P<0.05);0.036 25～150 μmol·L^-1侧柏酮明显促进其生长(P<0.05),9.4 μmol·L^-1时达增殖最高峰(37.03±0.52)%,且穿孔素表达率明显升高(39.94±1.04)%(P<0.05),余与空白组相比无显著差异. 结论: 9.4 μmol·L^-1侧柏酮明显促进人γδ T细胞增殖并上调其穿孔素的表达.     

柴胡疏肝散对免疫损伤性肝纤维化的防治作用

目的: 探讨柴胡疏肝散对肝纤维化的防治作用. 方法: 70只Wistar大鼠随机分为7组:正常组、病理模型组、秋水仙碱治疗组、柴胡疏肝散低、中、高剂量组、柴胡疏肝散预防组.除正常组外,其余各组均采用猪血清腹腔注射诱发肝纤维化,0.5 mL/只,2次/周,连续10周,5周后即可形成肝纤维化.预防组于造模同时给药(以柴胡疏肝散6.3 g·kg^-1),各治疗组于造模第6周给药,持续至第10周.秋水仙碱治疗组剂量0.5 mL·kg^-1·d^-1,柴胡疏肝散低、中、高剂量组(3.5,6.3,12.6 g·kg^-1·d^-1).用全自动分析法测定各组血清肝功能、放射免疫法测定肝纤维化指标,采用酸性水解法检测肝组织羟脯氨酸(hydroxyproline,HYP)含量,并观察病理变化. 结果: 与模型组比较,柴胡疏肝散中剂量组血清丙氨酸转氨酶(ALT),(156.5±3.2) U·L^-1 VS (78.4±4.6) U·L^-1(P<0.01);天冬氨酸转氨酶(AST),(387.4±12.7) U·L^-1 VS (232.5±13.5) U·L^-1(P<0.01);r-谷氨酰转肽酶(GGT),(128.2±4.0) U·L^-1 VS (78.5±4.2) U·L^-1(P<0.01);碱性磷酸酶(ALP),(152.75±5.01) U·L^-1 VS(42.51±4.67) U·L^-1(P<0.01);透明质酸(HA),(337.2±22.1)VS(140.6±21.2) μg·L^-1(P<0.01);PCⅢ(145.31±17.14) μg·L^-1VS(85.5±19.7)μg·L^-1(P<0.05);Ⅳ型胶原(135.1±22.4)VS(88.37±21.09)(P<0.05);层黏连蛋白(LN),(317.5±21.3)VS(148.4±18.4),(P<0.05)含量均显著下降, 肝组织(HYP)(527.2±34.2)VS(346.2±43.1)(P<0.01)含量显著降低. 结论: 柴胡疏肝散有明显的抗肝纤维化作用.     

藿香与广藿香抗氧化活性研究

目的: 研究藿香和广藿香的茎、叶提取物抗氧化活性,总多酚与总黄酮含量与抗氧化活性的关系,并分析二者药理药效是否具有可替代性. 方法: 采用DPPH法、ABTS法和FRAP方法进行研究. 结果: 发现总黄酮含量与抗氧化总能力有一定相关性(r=0.609 9).通过比较藿香与广藿香的抗氧化能力发现,广藿香乙酸乙酯部位清除DPPH自由基能力最强[IC₅₀(79.6±3.14) mg·L^-1],藿香乙酸乙酯部位清除ABTS自由基能力最强[IC₅₀ (4.41±0.23) mg·L^-1],广藿香石油醚部位Fe³⁺的还原能力最强[TEAC (1 822.99±1 141.13) μmol·g^-1]. 结论: 广藿香与藿香均具有一定抗氧化能力,且差别不大.     

新型乌贝速释片的生物等效性研究

目的: 制备一种新型乌贝速释片,并研究该剂型与乌贝散在Beagle犬体内的药代动力学和生物等效性。方法: 采取粉末直接压片法制备乌贝速释片,将6只Beagle犬随机分为2组,分别口服给予乌贝速释片和乌贝散,采用LC-MS/MS测定给药后不同时刻血浆中贝母素甲的浓度,采用DAS 2.0软件按非房室模型处理得药动学参数,评价速释片剂与散剂的生物等效性。结果: 乌贝速释片中贝母素甲的主要药动学参数为C_max(7.4±2.3) μg·L^-1,AUC_0-t (59.13±15.25) μg·L^-1·h^-1,T_max(1.5±0.0) h,乌贝散中贝母素甲的主要药动学参数为C_max(8.0±1.7) μg·L^-1,AUC_0-t (68.78±16.27) μg·L^-1·h^-1,T_max(1.5±0.0) h,乌贝速释片中贝母素甲的lnAUC_0-t和lnC_max的90%置信区间分别为乌贝散相应参数的95.4%~104.6%,90.9%~109.1%。结论: 自制的乌贝速释片与乌贝散具有生物等效性,且生产工艺简单,服用方便。     

北五味子乙素抗AngⅡ诱导的心肌成纤维细胞增殖及作用机制

目的：探讨北五味子木脂素（schisandra lignin,SCL）中的活性成分之一五味子乙素（schisandrin B,Sch B）对血管紧张素Ⅱ（angiotensinⅡ,AngⅡ）诱导心肌成纤维细胞（cardiac fibroblast,CFb）增殖的作用及作用机制。方法：以培养的新生Wistar大鼠乳鼠CFb为实验模型,实验分为对照组、AngⅡ模型（10^-1μmol·L^-1）组、Sch B 1,10,30 μmol·L^-1 3个剂量组。采用胰酶消化、差速贴壁法培养CFb,MTT法测定CFb增殖活力;分光光度计测定CFb中超氧化物歧化酶（SOD）活力和丙二醛（MDA）含量;流式细胞仪检测线粒体膜电位水平（mitochondrial membrane potential,MMP）ΔΨm;激光共聚焦显微镜检测细胞内钙离子水平。结果：AngⅡ组SOD 活力（122.72±21.59） U·mL^-1低于对照组（135.81±26.62）U·mL^-1,P<0.01,MDA水平（2.82±0.14） μmol·L^-1高于对照组（1.17±0.06） μmol·L^-1,P<0.01。SchB 1~30 μmol·L^-1能显著提高CFb中SOD活力,降低MDA含量,与AngⅡ相比（P<0.05或P<0.01）;AngⅡ组线粒体膜电位明显下降（587.6±55.2）ΔΨm,钙离子水平升高（335.6±28.3）,与对照组比较（3 069.2±282.4）ΔΨm,（50.7±4.6）,P<0.01或P<0.001,给予Sch B处理后ΔΨm升高（Sch B 10,30 μmol·L^-1）,钙离子水平下降（SchB 1~30 μmol·L^-1）,与AngⅡ组比较（P<0.05或P<0.01）。结论：Sch B通过提高CFb SOD活力、降低MDA水平、增加清除自由基能力抑制CFb增殖,其作用与其升高线性体膜电位水平,抑制细胞内钙离子浓度,减少促进CFb增殖的氧化物质生成有关。     

芪芍胶囊联合厄贝沙坦治疗糖尿病肾病

目的:探讨芪芍胶囊联合厄贝沙坦治疗糖尿病肾病(DKD)患者的疗效。方法:选取2014年1月至2015年4月期间河北省中医院收治的84例DKD患者为研究对象,并随机分为观察组和对照组,两组各42例。对照组给予厄贝沙坦治疗,150 mg qd,观察组在对照组治疗的基础上加用芪芍胶囊,5粒tid。比较两组治疗前后的糖代谢指标和肾功能指标,并比较两组患者的疗效。结果:治疗后观察组的空腹血糖(FPG),糖化血红蛋白(HbA1c)均显著低于对照组(P<0.05),治疗后观察组的血肌酐(SCr),尿蛋白排泄率(UAER),24 h尿蛋白(24 hUP),尿微量白蛋白(mALB),尿微量白蛋白与肌酐比(mALB/UCr),血尿素氮(BUN)分别为(135.7±18.9) μmol·L^-1,(155.3±28.1) mg·(24 h)^-1,(1.53±0.44) g,(39.7±12.9) μg·L^-1,(15.5±3.9) mg·g^-1,(6.59±0.87) mmol·L^-1,均显著低于对照组的(163.6±18.0) μmol·L^-1,(170.6±32.3) mg·(24 h)^-1,(1.79±0.52) g,(47.4±14.4) μg·L^-1,(17.7±3.8) mg·g^-1,(7.06±0.93) mmol·L^-1(P<0.05)。观察组的临床疗效明显优于对照组(P<0.05)。结论:与单用厄贝沙坦相比,芪芍胶囊联合厄贝沙坦治疗能更加显著地改善DKD患者的肾功能,延缓其进展,疗效显著,充分显示了中西医治疗的独特优势,值得临床推广应用。     

大豆异黄酮对去卵巢大鼠学习记忆和海马谷氨酸细胞及NR2B受体的影响

目的：通过建立H₂O₂诱导心肌细胞氧化应激损伤模型,观察番茄红素（lycopene）对心肌细胞的保护作用。方法：将原代培养的乳鼠心肌细胞随机分为正常组、模型组（H₂O₂）、番茄红素低剂量组（2.5 μmol·L^-1）、番茄红素中剂量组（5 μmol·L^-1）、番茄红素高剂量组（10 μmol·L^-1）。番茄红素与心肌细胞孵育4 h后,再加入H₂O₂,24 h后MTT法测定细胞存活率,比色法测定细胞培养上清液中乳酸脱氢酶（LDH）、肌酸激酶（CK）的含量和细胞超氧化物歧化酶（SOD）、过氧化氢酶（CAT）及丙二醛（MDA）含量,荧光电子显微镜下观察细胞核的形态变化,采用Western blot方法检测细胞凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）的变化。结果：与正常组比较,模型组心肌细胞存活率降低,心肌细胞LDH和CK释放量增加,SOD,CAT活性降低,MDA含量增加（P<0.01）,呈现出凋亡细胞的特征,Caspase-3的表达增高（P<0.01）。番茄红素高剂量组与模型组比较,心肌细胞的存活率增加（86.36±2.54）%,（54.15±10.97）%,P<0.01,H₂O₂所致乳鼠心肌细胞LDH释放量降低（319.53±23.48）,（495.27±31.57）U·L^-1,P<0.01,CK释放量降低（279.29±32.91）,（397.01±29.36）U·L^-1,P<0.01,SOD活性增高（47.94±2.64）,（37.19±5.78）U·mg^-1,P<0.01,CAT活性增高（34.95±4.67）,（16.35±4.84）U·mg^-1,P<0.01,MDA含量降低（14.27±2.43）,（20.11±2.09）μmol·g^-1,P<0.01,细胞核形态改善,Caspase-3的表达降低（1.40±0.20）,（1.94±0.22）,P<0.05。结论：番茄红素对H₂O₂所致乳鼠心肌细胞氧化应激损伤有明显保护作用。     

黄芩与五味子配伍对黄芩苷、五味子酯甲代谢动力学的影响

目的: 研究黄芩与五味子配伍对黄芩苷和五味子酯甲的大鼠药代动力学规律的影响。方法: 大鼠灌胃黄芩提取物2.5 g ·kg^-1﹑五味子提取物2.5 g ·kg^-1和黄芩提取物加五味子提取物各2.5 g ·kg^-1,分别于0.25,1,2,4,8,12,24 h取血分离血浆,采用HPLC测定其黄芩苷﹑五味子酯甲的含量,结果运用DAS 3.0计算其药动学参数。结果: 测定黄芩组黄芩苷的药动学参数分别为t1/2=(6.203±3.324)h,AUC=(41.868±28.254)μg ·L^-1 ·h^-1,C_max=(2.883±0.684) μg ·L^-1,MRT=(11.697±4.108) h,V_d=(568 112.69±81 364.658)L ·kg^-1,CL=(98 526.569±93 774.892)L ·h^-1 ·kg^-1;黄芩+五味子组黄芩苷的药动学参数分别为t1/2=(10.686±1.533)h,AUC=(53.064±30.047) μg ·L^-1 ·h^-1,C_max=(3.168±1.312) μg ·L^-1,MRT=(16.147±1.79) h,V_d=(2 240 724.1±1 824 718.9)L ·kg^-1,CL=(139 855.27±107 995.59)L ·h^-1 ·kg^-1;黄芩+五味子组五味子酯甲的药动学参数分别为t_1/2=(21.544±14.611)h,AUC=(11.554±5.516) μg ·L^-1 ·h^-1,Cmax=(0.311±0.074) μg ·L^-1,MRT=(34.139±18.532) h,V_d=(12 153 917±5 806 489.8)L ·kg^-1,CL=(564 758.71±384 128.86)L ·h^-1 ·kg^-1;五味子组五味子酯甲的药动学参数分别为t1/2=(4.926±5.371)h,AUC=(4.988±3.029) μg ·L^-1 ·h^-1,C_max=(0.287±0.071) μg ·L^-1,MRT=(12.002±6.854) h,V_d=(3 091.656±1 585.602)L ·kg^-1,CL=(615.571±250.643)L ·h^-1 ·kg^-1。结论: 黄芩与五味子配伍,可以促进黄芩苷和五味子酯甲在血液中的吸收,并可延长这两种成分的半衰期,使其血药浓度维持在较高水平,达到协同增效的目的。     

列当属、肉苁蓉属和草苁蓉属植物传统药物学调查

通过野外资源调查、整理各大标本馆标本原始记录和查阅文献记载等方法,系统整理、总结、归纳了列当属、肉苁蓉属和草苁蓉属民族药用植物种类、功效及民间使用情况,结果表明列当属6种药用植物在4个少数民族间作为7种民族药应用,草苁蓉属2种药用植物在8个少数民族间作为10种民族药应用,肉苁蓉属2种药用植物在3个少数民族间作为3种民族药应用,且同种药用植物常作不同民族药;发现3属植物的传统疗效主要集中在补肾壮阳、止血和止痛3个方面,并且该传统疗效与现代药理研究结果基本吻合。因此深入研究植物种类丰富的列当属植物资源对缓解肉苁蓉植物资源匮乏局面和扩大药源具有积极意义。     

厚朴与凹叶厚朴群体遗传学研究

目的:对厚朴与凹叶厚朴的群体遗传学进行研究,为中药厚朴的质量控制提供分子生药学方面的依据。方法:对厚朴与凹叶厚朴15个居群应用2个叶绿体基因间序列psbA-trnH和trnL-trnF进行PCR扩增并测序,计算厚朴与凹叶厚朴单倍型频率,用程序HaploNst分析遗传多样性和遗传结构,应用TCS version 1.13软件构建单倍型网状进化树。结果:厚朴与凹叶厚朴均无特有单倍型存在,但单倍型频率存在显著差异,已开始出现遗传分化的趋势,NST略大于GST。结论:厚朴与凹叶厚朴在遗传上已出现遗传分化的趋势,但尚未完全分化成彼此独立的单系。     

追风伞中黄酮类成分的研究

目的:对追风伞中的黄酮类成分进行分离、鉴定。方法:追风伞干燥全草用95%乙醇溶液加热回流提取,减压回收乙醇,浓缩液依次用石油醚、氯仿萃取后,水层部分利用大孔吸附树脂、硅胶柱色谱、反相Rp-18柱色谱及重结晶等方法进行分离及纯化,并通过1H-NMR,13C-NMR,EI-MS及理化常数对分离化合物进行结构鉴定。结果:从追风伞提取物中分离鉴定9个黄酮类化合物,分别为:木犀草素(1),木犀草素-4′-O-β-D-葡萄糖苷(2),刺槐素-7-O-β-D-葡萄糖苷(3),芦丁(4),刺槐素(5),槲皮素(6),槲皮素-3-O-β-D-葡萄糖苷(7),山柰酚-3-O-β-D-葡萄糖苷(8),异鼠李素-3-O-β-D-葡萄糖苷(9)。结论:所得化合物均为首次从追风伞中分离鉴定。     

Antimalarial and safety evaluation of Pluchea lanceolata (DC.) Oliv. & Hiern: In-vitro and in-vivo study

Ethnopharmacological relevance

Many of the effective therapeutic strategies have been derived from ethnopharmacologically used natural products. Pluchea lanceolata is an herb employed in Indian folk medicine for malaria like fever but it lacks proper pharmacological intervention.

Aim of the study

To evaluate antimalarial and safety profile of Pluchea lanceolata: an in-vitro, in-vivo for its ethnopharmacological validation.

Materials and methods

Methanol, butanol, ethyl acetate, chloroform, hexane extracts and its isolate, taraxasterol acetate (TxAc) were obtained from air dried aerial part of Pluchea lanceolata. These were tested in-vitro against chloroquine-sensitive strain of Plasmodium falciparum NF54 by measuring the parasite specific lactate dehydrogenase activity. The most potent hexane extract and TxAc were further validated for in-vivo antimalarial and safety evaluation. TxAc, a pentacyclic-triterpene isolated from the most active fraction was further evaluated with special emphasis on inflammatory mediators involved in malaria pathogenesis. Murine malaria was induced by intra-peritoneal injection of Plasmodium berghei infected red blood cells to the male Swiss inbred mice. Mice were orally treated following Peters 4-Day suppression test. In-vivo antimalarial efficacy was examined by evaluating the parasitaemia, percent survival, mean survival time, blood glucose, haemoglobin and pro-inflammatory mediators involved in malaria pathogenesis.

Results

Hexane extract and TxAc showed promising antimalarial activity in-vitro and in-vivo condition. TxAc attributed in inhibition of the pro-inflammatory cytokines as well as afford to significant increase in the blood glucose and haemoglobin level when compared with vehicle treated infected mice. We have not observed the synergistic action of combinations of chloroquine and TxAc from our experimental results. In-vitro and in-vivo safety evaluation study revealed that hexane extract is non toxic at higher concentration.

Conclusion

Present study further validates the ancient Indian traditional knowledge and use of Pluchea lanceolata as an antimalarial agent. Study confirms the suitability of Pluchea lanceolata as a candidate for further studies to obtain a prototype for antimalarial medicine.     

In vitro and in vivo anti-Helicobacter pylori activity of Calophyllum brasiliense Camb.

Aims of the study

Calophyllum brasiliense (Camb.) is a medicinal tree that grows particularly in the hilly and forested regions of Brazil. Preparations from its stem bark are popular remedies for the treatment of chronic ulcers. Since earlier investigations on bark extracts evidenced gastroprotective and gastric acid inhibitory properties, this study evaluated the effects of hydroethanolic extract (HEECb) and the dichloromethanic fraction (DCMF), from Calophyllum brasiliense stem bark, against Helicobacter pylori, in vitro and in vivo.

Materials and methods

The in vitro assays were performed using the disk diffusion and broth microdilution methods to determine the minimum inhibitory concentration (MIC) values. The test substances were evaluated in vivo taking into account the delay in the gastric ulcer healing in Wistar rats, infected with Helicobacter pylori.

Results

DCMF appeared the most active and potent in vitro against Helicobacter pylori growth with an MIC of 31 μg/mL. In the in vivo assays, rats ulcerated by acetic acid, and inoculated with Helicobacter pylori showed a marked delay in ulcer healing. Treatment with HEECb (50, 100 and 200 mg/kg) and DCMF (100 and 200 mg/kg) reduced the ulcerated area in a dose-dependent manner. While DCMF, at 200 mg/kg, increased the prostaglandin E₂ (PGE₂) level, both HEECb and DCMF decreased the number of urease-positive animals, as confirmed by the reduction of Helicobacter pylori presence in histopathological analysis.

Conclusion

The results suggest that the antiulcer activity of Calophyllum brasiliense is due, in part, to its anti-Helicobacter pylori action, validating the popular use of this species.     

暗紫贝母、栽培瓦布贝母及浙贝母药效学比较

目的:比较暗紫贝母、栽培瓦布贝母及浙贝母镇咳、祛痰及平喘作用,明确栽培瓦布贝母作为野生川贝母替代资源的可行性.方法:采用小鼠氨水引咳及豚鼠枸橼酸引咳法比较3种贝母的镇咳作用.小鼠酚红排痰法及大鼠毛细管排痰实验法比较3种贝母的祛痰作用;整体动物引喘实验法比较3种贝母的平喘作用.每个实验均分为5个组,分别为阴性对照组,阳性对照组,暗紫贝母组,栽培瓦布贝母组及浙贝母组.阴性组采用蒸馏水或生理盐水灌胃5d.镇咳实验中3种贝母的剂量分别为2.5g·kg-1·d-1(小鼠),1.5g·kg-1·d-1(豚鼠);祛痰实验中3种贝母的剂量分别为2 g·kg-1·d-1(小鼠),1 g·kg-1·d-1(大鼠),平喘实验中3种贝母的剂量为1.5 g·kg-1·d-1.结果:栽培瓦布贝母祛痰作用与暗紫贝母相当,小鼠酚红排泌量与阴性对照组比较,P＜0.01,大鼠毛细管排泌量与阴性对照组比较P＜0.05,栽培瓦布贝母平喘作用与暗紫贝母相当,哮喘潜伏期与阴性对照组比较,P＜0.05,暗紫贝母与栽培瓦布贝母作用差异无统计学意义;栽培瓦布贝母镇咳作用与暗紫贝母及浙贝母相当,减少枸橼酸引咳后5 min内咳嗽次数与阴性组比较,差异有统计学意义.结论:栽培瓦布贝母可作为野生川贝母的替代资源推广种植与应用.     

In vitro activity of Tridax procumbens against promastigotes of Leishmania mexicana

Ethnopharmacological relevance

Tridax procumbens is an active herb against leishmaniasis.

Aim of the study

Leishmaniasis is a group of diseases caused by Leishmania protozoa. We investigated the antileishmanial activity of Tridax procumbens extracts and a pure compound against promastigotes of Leishmania mexicana, the causative agent of cutaneous leishmaniasis in the New World.

Materials and methods

Extracts and (3S)-16,17-didehydrofalcarinol (1) were obtained by chromatographic methods from Tridax procumbens, and the latter identified by spectroscopic analysis. The effect of these extracts and 1 on the growth inhibition of promastigotes of Leishmania mexicana was evaluated. In order to test the safety of extracts and 1, mammalian cells were treated with them, and cell viability was assessed using trypan blue and MTT.

Results

We demonstrated that extracts of Tridax procumbens and 1 showed a pronounced activity against Leishmania mexicana. The methanol extract inhibited promastigotes growth of Leishmania mexicana with a 50% inhibitory concentration (IC₅₀) of 3 μg/ml, while oxylipin 1 exhibited the highest inhibition at IC₅₀ = 0.478 μg/ml.

Conclusions

In this study we report the biological activity of extracts and (3S)-16,17-didehydrofalcarinol (1), obtained from Tridax procumbens, on the promastigote form of Leishmania mexicana, with no effect upon mammalian cells.     

Anthelmintic activity of Arisaema franchetianum and Arisaema lobatum essential oils against Haemonchus contortus

Ethnopharmacological relevance

Arisaema franchetianum and Arisaema lobatum are two perennial plants native to China. Arisaema franchetianum is universally used to promote the subsidence of induration and swelling, quicken blood and relieve pains, and kill intestinal parasites in humans and animals. Arisaema lobatum is used to treat malaria, intestinal parasites, and snake and insect bites in humans and animals. The aim of this study was to determine the composition of the essential oils from Arisaema franchetianum and Arisaema lobatum and evaluate the anthelmintic effect against Haemonchus contortus.

Materials and methods

Two oils were investigated by GC and GC–MS. The anthelmintic bioassay tests of Arisaema franchetianum and Arisaema lobatum essential oil, linalool and carvacrol were performed using egg hatch assay (EHA), larval development assay (LDA) and larval migration inhibition assay (LMIA).

Results

Fifty six components representing 96.88% of the Arisaema franchetianum oil and 64 components representing 96.88% of the Arisaema lobatum oil were identified. Carvacrol and linalool were found to be the major constituents of two oils. In the EHA, greater than 99% inhibition were observed with Arisaema franchetianum oil at 10 mg/mL (CE₅₀ 1.63 mg/mL) and Arisaema lobatum oil at 5 and 10 mg/mL (CE₅₀ 0.48 mg/mL). In the LDA, both oils induced complete inhibition at 10 mg/mL, with the CE₅₀ being 1.10 mg/mL for Arisaema franchetianum oil and 0.73 mg/mL for Arisaema lobatum oil. In the LMIA, the Arisaema franchetianum oil and Arisaema lobatum oil at best inhibited 74.1% and 95.6% of larval migration at 10 mg/mL, respectively. Carvacrol exhibited similar activity to Arisaema lobatum essential oil and linalool did not show high activity in every assay.

Conclusions

These data show for the first time that the essential oils obtained from Arisaema franchetianum or Arisaema lobatum had promising anthelmintic activity against Haemonchus contortus. Arisaema plant may offer an alternative source for the control of gastrointestinal nematodes of sheep and goats.     

丛枝菌根真菌与哈茨木霉菌合用对连作丹参生长及质量的影响

该文探讨田间条件下丛枝菌根真菌(AMF)与哈茨木霉菌合用对丹参生长及质量的影响。采用田间小区试验来进行研究,使用HPLC测定丹参地上部分和地下部分有效成分含量,通过形态观察并计算根部发病率,并结合方差统计学方法对各指标的测定结果进行分析。结果显示,单独接种AMF以及AMF与哈茨木霉菌合用能够有效的降低连作丹参根部病害的发生率,其中AMF与哈茨木霉菌合用效果更佳,比CK组降低了61.50%。几种处理对丹参生物量影响没有显著差异,但都能提高丹参根部各种有效成分的含量。单独接种AMF以及AMF+哈茨木霉都能显著提高丹参根中丹酚酸B的含量(P<0.05),其中AMF对丹酚酸B促进作用最明显,提高了27.97%;接种AMF能显著提高丹参根丹参酮I和丹参酮ⅡA的含量(P<0.05),其中丹参酮I约提高了70.14%;丹参酮ⅡA约提高了51.42%。另外,单独接种AMF以及AMF+哈茨木霉均能显著提高丹参根中隐丹参酮的含量,提高了约30.67%。从而得出单独接种AMF以及AMF+哈茨木霉能够有效减低连作丹参病害的发生率,同时提高连作丹参药材的质量。     

金银花类药用植物IspE和IspH基因克隆和生物信息学分析

目的:克隆金银花类药用植物4-二磷酸胞苷-2-C-甲基赤藓糖激酶(4-diphosphocytidyl-2-C-methyl-D-erythritol kinase,IspE)和4-羟基-3-甲基-2-邻苯基二磷酸还原酶(4-hydroxy-3-methylbut-2-enyl diphosphate reductase,IspH)基因,并对其基因序列、蛋白特性和转录活性进行分析、比较.方法:从忍冬Lonicera japonica转录组测序结果中分析获得了IspE,IspH基因.分别以忍冬、红白忍冬L.japonica var.chinensis、红腺忍冬L.hypoglauca和水忍冬L.dasystyla新鲜花蕾为材料,利用RT-PCR技术克隆获得了4种金银花类药用植物IspE和IspH基因的全长cDNA.运用生物信息学分析软件,预测编码蛋白的结构和功能,并通过RT-PCR检测IspE和IspH在忍冬、红白忍冬、红腺忍冬、水忍冬花蕾中的转录情况.结果:金银花类药用植物IspE基因含有1个完整的开放阅读框,长度为1 221 bp,编码406个氨基酸;IspH含有一个完整的开放阅读框,长度为1 380 bp,编码459个氨基酸.IspE和IspH均为非分泌蛋白,均定位于叶绿体中.RT-PCR分析结果表明在忍冬、红腺忍冬和水忍冬的花蕾中IspE,IspH基因的转录水平没有显著差异,但红白忍冬花蕾中IspE,IspH基因的转录水平均显著高于忍冬.结论:克隆获得忍冬、红白忍冬、红腺忍冬和水忍冬中IspE,IspH基因,并证实了其在不同金银花类药用植物中的表达,为进一步研究IspE,IspH基因对萜类化合物生物合成和花香气以及颜色的影响奠定了基础.