线性探针微透析研究阿魏酸经皮给药后药动学

目的:建立线性探针测定阿魏酸皮下浓度方法,考察温度、灌流液流速、浓度对线性探针回收率的影响,研究阿魏酸溶液经皮给药后体内药动学.方法:采用减量法测定阿魏酸线性探针的体内外同收率的稳定性,采用增量法测定体外回收率的影响因素,选择CD-1裸鼠作为实验动物,采用经皮微透析法进行体内药动学研究,HPLC测定透析液中阿魏酸的浓度.结果:温度对阿魏酸体外同收率有显著性影响,随着温度的升高,阿魏酸的体外回收率显著提高,随着流速的增加阿魏酸的体外同收率呈指数下降,在测定范围内,浓度对阿魏酸的体内外回收率无影响,在测定时间内阿魏酸体内外同收率稳定性、重复性良好,体外平均回收率为(24.82±1.01)％;体内平均回收率为(16.50±1.92)％;经皮给药后,阿魏酸86 min达到峰值,平均滞留时间为291 min.结论:线性探针可用于阿魏酸经皮给药的研究;阿魏酸可迅速透过角质层,适宜制成经皮给药制剂.     

阿魏酸体内外微透析回收率及其乳剂的经皮皮肤药动学研究

目的 研究阿魏酸在自制线性微透析探针上的体内外回收率与灌流速度、药物浓度之间的关系,并对自制阿魏酸乳剂在大鼠上进行经皮皮肤药动学研究.方法 采用HPLC测定透析液中阿魏酸浓度,考察不同灌流速度、不同药物浓度对探针体外正、反向回收率和体内反向回收率的影响,采用微透析技术进行经皮皮肤药动学研究.结果 自制探针性质稳定;在所...     

阿魏酸、川芎及复方脑得生中阿魏酸在大鼠体内的药动学研究

目的:建立RP-HPLC测定大鼠血浆中阿魏酸浓度的方法,研究阿魏酸、川芎及复方脑得生片中阿魏酸的药动学特点,评价中药材中其他成分和复方中其他配伍对阿魏酸药动学的影响.方法:大鼠分别灌胃给予阿魏酸对照品、川芎药材提取物、复方脑得生片,断尾采血,离心,取血浆适量,加甲醇沉淀蛋白,取上清液过滤,用HPLC分析,以C_(18)为固定相,流动相为甲醇-0.5%醋酸水溶液(30:70),在320 nm检测阿魏酸血药浓度,3p97软件处理数据.结果:阿魏酸血浆在0.05～1 mg·L~(-1)线性关系良好,血浆中最低检测质量浓度为13μg·L~(-1).大鼠灌胄给予阿魏酸对照品、川芎提取物和脑得生片后的药-时曲线均符合二房室模型,主要药动学参数AUC_(0～t),C_(max),CL在阿魏酸对照品、川芎提取物和脑得生片各组间均有统计学意义.结论:本实验建立的RP-HPLC测定法,专属、准确、灵敏,适用于阿魏酸在大鼠体内的药动学研究.川芎药材中的其他组分会影响阿魏酸的口服吸收,而复方配伍后可促进川芎中阿魏酸的吸收,提高阿魏酸的生物利用度.     

邓氏冠心止痛巴布剂中阿魏酸的透皮行为研究

目的研究邓氏冠心止痛巴布剂中阿魏酸的体外经皮渗透性。方法采用改良Franz扩散池法,生理盐水为吸收液,HPLC测定经皮渗透的阿魏酸量,研究邓氏冠心止痛巴布剂中阿魏酸的经皮渗透行为。结果邓氏冠心止痛膏巴布剂中阿魏酸的经皮渗透是由皮肤控制的。在1～18 h内,阿魏酸经大鼠皮的透皮速率常数为14.015μg.cm-2.h-1;在1～9 h内,阿魏酸经小鼠皮肤透皮速率常数为20.541μg.cm-2.h-1;在48 h内,阿魏酸经大鼠皮的平均相对渗透率为26.16%;经小鼠皮的平均相对渗透率为23.44%。结论邓氏冠心止痛巴布剂有较好的经皮渗透性能和缓释效果。     

大鼠口服川芎和通脉方后阿魏酸的药动学比较

目的:比较大鼠灌服川芎提取物和通脉方后阿魏酸的药代动力学特征差异。方法:Wistar大鼠分别灌胃给予川芎提取物和通脉方(相当于阿魏酸单体9.0 mg.kg-1)后,于不同时间点采集大鼠血浆。血浆样品经乙酸乙酯萃取后,用高效液相色谱法测定阿魏酸的血药浓度,并用3P97药动学软件对其血药浓度-时间数据进行处理。结果:阿魏酸在大鼠体内的药代动力学行为符合开放一室模型,川芎提取物和通脉方中阿魏酸Ke分别为(0.015±0.002),(0.008±0.001)min,t1/2(Ke)分别为(46.850±5.414),(87.033±11.025)min,存在显著性差异(P<0.01);AUC分别为(328.440±115.461),(625.718±139.798)μg.min-1.mL-1,CL/F(s)分别为(0.032±0.013),(0.015±0.004)L.kg-1.min-1,存在显著性差异(P<0.05)。结论:丹参、葛根与川芎配伍使用,可以降低阿魏酸的消除速率,延长其体内作用时间和提高生物利用度。     

小儿咳喘贴黑膏药体外经皮渗透行为研究

目的研究小儿咳喘贴黑膏药体外透皮吸收特性,从渗透屏障和渗透介质角度考察其体外经皮渗透的最佳条件。方法以芥子碱硫氰酸盐为指标,HPLC测定小儿咳喘贴黑膏药透皮接收液中有效成分。采用改良的Franz扩散池法,分别以蒸馏水、生理盐水、磷酸盐缓冲液为渗透介质,大鼠、小鼠、兔和裸鼠腹皮为渗透屏障,考察其体外经皮渗透的最佳条件。结果以裸鼠腹皮为渗透屏障,以磷酸盐缓冲液为渗透介质时效果最佳。芥子碱硫氰酸盐自小儿咳喘贴黑膏药中体外释放的方程为QR=0. 3254t+0. 1485,r=0. 9966,释放速率为0. 3254μg/(cm2·h),透皮特性方程为QT=0. 5147t+0. 476,r=0. 9969,透皮速率0. 5147μg/(cm2·h)。结论小儿咳喘贴黑膏药中,芥子碱硫氰酸盐的体外透皮行为符合零级动力学方程,芥子碱硫氰酸盐的释放速率小于透皮速率,表明小儿咳喘贴黑膏药是通过控制药物释放达到长效控释的目的,不同皮肤及渗透介质对其体外渗透行为有影响。     

薄荷脑对雪上一枝蒿甲素经皮促渗的药动学及体内外相关性研究

目的:考察不同浓度的薄荷脑对雪上一枝蒿甲素经皮促渗的药动学及体内外相关性。方法:分别制备含1%、3%、5%薄荷脑的雪上一枝蒿总碱凝胶,采用改良的Franz扩散池法进行体外释放及透皮研究;结合UPLC-MS/MS联用技术检测雪上一枝蒿甲素在小鼠血液中的药物浓度随时间的变化过程,并对其进行体内药动学研究;以体外24h溶出率为横坐标,AUC为纵坐标,对体外透皮结果与体内药动学结果进行回归分析。结果:与未加薄荷脑的雪上一枝蒿总碱凝胶相比,1%、3%、5%三种不同浓度薄荷脑的增渗倍数(ER)分别为2. 62、3. 26、4. 48,促渗雪上一枝蒿甲素的相对生物利用度分别为163. 39%、189. 16%、310. 69%,体内外相关系数为0. 9591。结论:体内外促渗效果一致,此方法可用于评价透皮给药系统体内外的相关性。     

黄芩苷透皮给药系统的体内药代动力学研究

目的 研究黄芩苷透皮给药系统的体内药代动力学.方法 采用高效液相色谱法测定家兔血清中的黄芩苷透皮给药系统中黄芩苷及在兔体内的药代动力学参数.结果 血清药物浓度在1.5～50 μg/ml范围内线性关系良好,体内药代动力学呈一级吸收的二室模型.结论 黄芩苷透皮给药系统在家兔体内的药代动力学参数的变化表明其药动学符合二室模型,该实验可为临床用药提供药动学参数.     

血液微透析技术分析阿魏酸口服给药后药代动力学过程

目的:应用血液微透析技术测定阿魏酸口服给药后药代动力学参数.方法:采用浓差法(增量法和减量法)考察探针回收率的日内稳定性及灌流速度、阿魏酸质量浓度、温度对探针回收率的影响,通过零净流量法测定阿魏酸体外回收率,并将结果与浓差法所测回收率进行比较.采用血液微透析法进行体内药动学研究,HPLC测定透析液中阿魏酸含量.结果:探针回收率日内稳定性良好;体外回收率与阿魏酸质量浓度无关,与灌流速度成指数型负相关;温度越高,回收率越高;利用零净通量法能准确地测定透析介质中阿魏酸含量及回收率;增量法、减量法及零净通量法测定的回收率一致.在2.5 μL·min-1流速下,体内平均回收率(28.69±1.95)％,口服给药后阿魏酸9.33 min达峰值,消除半衰期12.52 min.结论:微透析取样技术可用于阿魏酸的药动学研究,减量法可作为体内微透析研究中阿魏酸回收率的测定方法.     

单方与复方给药后洋川芎内酯I在大鼠体内的药动学比较研究

目的:建立LC-MS法测定大鼠血浆洋川芎内酯I(senkyunolide I,SI)的浓度,比较活络效灵丹加减方(HLXL)和川芎单味药材提取物中SI在大鼠体内的药代动力学参数有无显著性变化,评价HLXL提取物中其他成分对SI药动学的影响.方法:将12只大鼠随机分成2组,按含SI计4.53 mg· kg-1分别灌胃给予HLXL提取物和川芎提取物,不同时间点采血后进行LC-MS分析,测定血浆SI浓度,采用DAS 2.0软件对主要药动学参数进行计算,并用SPSS 16.0软件对其进行独立样本t-检验和非参数t-检验分析.结果:SI在6.750～675.0 μg·L-1线性关系良好,最低检测限为6.750 μg·L-1.大鼠灌胃HLXL提取物和川芎提取物的药-时曲线均符合二房室模型,测得SI的Cmax,AUCo-24h和CL有显著性差异,其他参数均无显著性差异.结论:该实验建立的LC-MS测定法专属、准确、灵敏,适用于HLXL提取物和川芎提取物中SI的药动学研究.实验结果表明,HLXL中其他药材对SI的吸收有较大影响.     

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。     

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.     

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。     

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.     

北柴胡糖基转移酶基因BcUGT3,BcUGT6的表达分析及其原核表达

该研究测定了北柴胡中2个糖基转移酶基因BcUGT3和BcUGT6在不同组织中的转录水平及MeJA处理后不定根中的转录水平。BcUGT3在根、叶、花和果中的转录水平无明显差异,但均高于茎中的转录水平。BcUGT6在叶中转录水平最高,在花中最低。以未处理的为对照,BcUGT6在MeJA处理后2 h,8 h,24 h,2 d,4 d的北柴胡不定根中,转录水平均提高近2倍,表明BcUGT6的转录受MeJA影响较小。BcUGT3转录水平随处理时间的延长不断增加,4 d时,达7倍左右。利用载体pET-28a（+）,进行了2个基因的原核表达。IPTG诱导后,宿主菌表达出了目的蛋白,并获得了纯化蛋白。为后续开展这2个基因的功能研究奠定了基础。     

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.