微量热法研究黄连及其主要组分配伍的抑菌作用

基于微量热法,研究黄连、黄连的主要组分小檗碱、药根碱、巴马汀及其配伍模拟方的抑菌作用.以HPLC法测定黄连中小檗碱、药根碱和巴马汀的含量,并根据其含量比值配伍模拟方;微量热法测定黄连、小檗碱、药根碱、巴马汀及其模拟方对痢疾杆菌的生长代谢曲线,得出相应的热动力学参数,并进行对应分析.结果表明黄连、小檗碱、药根碱、巴马汀及其模拟方对痢疾杆菌的生长代谢均有不同程度的抑制作用,黄连作用最强,单体生物碱作用弱,配伍模拟方作用增强,但并未显现明显协同作用,黄连的抑菌作用可能为多种活性成分的综合作用.     

微量热法比较黄连中四种生物碱的抗菌作用

采用微量热法研究和比较中药黄连中四种生物碱(BAs)的抗菌(大肠杆菌)作用。这四种生物碱分别为小檗碱、黄连碱、巴马汀和药根碱。用LKB-2277生物活性检测系统,以停留法测定了37 ℃时大肠杆菌在BAs作用下的热流功率-时间曲线,并记录生长速率常数k, 指数生长期和稳定期的最大热功率Pm, log、Pm, stat, 生长抑制率I, 传代时间tg,指数生长期的总产热量Qt, log,半数抑菌浓度IC50等热动力学参数。结果显示这四种生物碱具有相异的抗大肠杆菌生长代谢的作用,k, Pm, log和Qt, log值随BAs浓度的增加而相应的减少。综合分析k, Pm, log, Qt, log, I 和IC50值可以看出,BAs抑制大肠杆菌生长代谢强度按小檗碱、黄连碱、巴马汀、药根碱的顺序依次减弱。构效关系研究表明,母环C-2和C-3上连接的亚甲二氧基比甲氧基更能显著增强相应化合物的抗菌作用,而C-9和C-10上连接的亚甲二氧基或甲氧基对这种抗菌作用影响不大。     

左金丸及类方HPLC指纹图谱与生物热活性的“谱-效”关系研究

HPLC法建立左金丸及类方水提液的特征指纹图谱, 计算各特征峰的相对峰面积, 并采用中药色谱指纹图谱相似度评价系统A版(2004 A)计算类方间的相似度; 微量热法测定大肠杆菌在左金丸及类方水提液作用下的热谱曲线, 得到相应的热动力学参数, 如生长速率常数k, 最大产热功率Pm, 最高峰的出峰时间tm和总产热量Qt等, 并对这些热动力学参数进行主成份分析. 典型相关分析法对左金丸及类方HPLC 指纹图谱中特征峰的相对峰面积与其类方作用下大肠杆菌生长代谢的主要热动力学参数相关联, 研究“谱-效”相关性. 结果表明, HPLC 指纹图谱与生物热活性存在很好的相关性, 类方中吴茱萸次碱、盐酸巴马汀、盐酸小檗碱的含量差异是导致其生物热活性不同的主要原因.     

卤代邻羟苄基谷氨酸(酯)的合成、表征及抑菌活性

分别用氯代水杨醛、溴代水杨醛与L-谷氨酸及其酯反应,首先制得水杨醛缩谷氨酸类席夫碱(2),再经硼氢化钠还原制得邻羟苄基取代谷氨酸及其酯(3),化合物的结构经IR、1H NMR、MS与元素分析表征确认. 测试表明,在质量浓度为0.05%时,卤代邻羟苄基谷氨酸对大肠杆菌具有高的抑菌活性,抑菌率高达97%,对白色念珠菌、金黄色葡萄球菌的抑菌效果较差,但其甲酯化产物卤代邻羟苄基谷氨酸甲酯对白色念珠菌具有高的抑菌活性,抑菌率高达100%,对大肠杆菌、金黄色葡萄球菌的抑菌效果较差,说明该类衍生物对菌株的抑杀作用具有明显的专一性和特异性,氯代邻羟苄基谷氨酸甲酯、溴代邻羟苄基谷氨酸甲酯是白色念珠菌的优良抗菌化合物。     

微量量热法研究黄连中小檗碱类生物碱对金黄色葡萄球菌生长代谢的影响

采用微量量热法研究中药黄连中具有抑菌作用的3种小檗碱类生物碱成分对金黄色葡萄球菌生长代谢的影响．在不同给药条件下,以表达功率．时间曲线（热谱曲线）的特征参数生长速率常数k、半数抑菌浓度（IC50）、最大输出功率（Pmax）及达峰时间（tp）和总产热量（Qt）为指标,对3种小檗碱类生物碱成分抑制金黄色葡萄球菌生长代谢程度进行客观地量化评价．结果表明,k、Pmax和Qt均随3种小檗碱类生物碱成分浓度的增加而相应的减小;tp随3种小檗碱类生物碱成分浓度的增加而相应的延长;其IC50分别为小檗碱101．4μg／mL、巴马汀241．0μg／mL和药根碱792．3μg／mL,即抑菌活性大小顺序为小檗碱〉巴马汀〉药根碱．初步构效关系研究提示,位于苯环的C2、C3上的亚甲二氧基或甲氧基较羟基更有助于提高其抑菌活性．     

微量热法测定细菌的最低生长温度

微量热法测定细菌的最低生长温度南照东，刘永军，孙海涛，张洪林（曲阜师范大学化学系曲阜２７３１６５）关键词微量热法，细菌的生长热谱，最低生长温度，生长速率常数我们应用微量热法测定了大肠杆菌（Ｅ．ｃｏｌｉ），白色葡萄球菌（Ｓ．ａｌｂｕｓ），金黄色葡萄球菌...     

用微量热法研究左金丸及类方药的药性差异

采用微量热法测定了大肠杆菌在左金丸、甘露散、茱萸丸和反左金丸水煮液作用下的生长热谱曲线,获得相应的生物热动力学参数,经主成份分析并结合本草文献报道,综合分析左金丸及类方的药性差异,以从生物物理和生物化学的角度,探讨中医经典类方药性差异的客观真实性.实验发现,左金丸及类方的水煮液对大肠杆菌的生长代谢表现出不同的活性作用.寒凉药方左金丸和甘露散不同程度的抑制细菌的生长代谢.温热药方反左金丸和茱萸丸对细菌的生长代谢有不同程度的促进作用.类方之间存在较稳定的药性差异.类方药性主要受生物热动力学参数:第二指数生长期的生长速率常数k2和最大发热功率Pm2的影响.     

微量热法研究鹿茸4个活性部位对肠道特征菌群生长代谢的影响

建立灵敏表征鹿茸中具有补益活性的4个部位对两种肠道特征菌群生长代谢影响的评价方法.采用微量热法,在不同给药条件作用下,以产热功率-时间曲线(热谱曲线)的特征参数(生长速率常数(k)、最大产热功率(Pmax)、达峰时间(tp)及有效率(E)等)为指标,对肠道主要有益菌(青春双歧杆菌)生长代谢程度进行客观地量化评价.挑选鹿茸促菌最强的活性部位,进一步考察该部位对病原微生物(金黄色葡萄球菌)生长抑制的作用规律.结果表明,鹿茸不同活性部位促进青春双歧杆菌生长代谢强弱顺序为:正丁醇乙醚氯仿乙酸乙酯.活性最强的正丁醇萃取液对病原微生物(金黄色葡萄球菌)抑制影响的研究表明,正丁醇萃取液在浓度大于200μg·mL-1时,对金葡菌生长具有抑制作用.且tp随药液浓度的增加而相应地延长;k,Pmax均随浓度的增加而相应地减小.微量热法具有灵敏准确、实时在线、重现性好等特点,可用于名贵药材——鹿茸的活性部位的筛选,也为药效物质的研究提供了一个新的研究思路和方法.     

茜素红荷移分光光度法测定酮康唑含量

酮康唑为合成的咪唑二烷衍生物,对皮真菌、酵母菌(包括念珠菌属、糠秕孢子菌属、球拟酵母菌属、隐球菌属)、双相真菌和真菌纲具有抑菌和杀菌活性,临床用于治疗深部和浅部真菌病,包括胃肠道真菌感染、局部用药无效的阴道白色念珠菌病、皮肤真菌感染、白色念珠菌和球孢子菌等。目前,已报道的     

黄柏生物碱成分在二妙丸类方中保护尿酸盐致人血管内皮细胞损伤作用及其配伍变化的研究

研究对二妙丸类方(二妙丸、三妙丸、四妙丸和加味四妙丸)起重要作用的基本方二妙丸的物质基础及其配伍变化. 正交组合二妙丸类方组方药有效部位, 四氮唑盐(MTT)法评价优选出对保护尿酸钠结晶(MSU)致人血管内皮细胞(HUVEC)损伤起重要作用的二妙丸黄柏生物碱部位, HPLC/DVD法建立黄柏生物碱部位指纹图谱, HPLC-ESI-MS/MS分析共有峰成分, 鉴定了12个生物碱成分, 并比较了二妙丸类方有效部位群HPLC/DVD指纹图谱配伍类方前后黄柏生物碱共有峰相对峰面积比值的变化. 结果表明, 黄柏生物碱配伍二妙丸及其类方后总相对含量分别为122.9%, 80.7%, 60.2%和94.2%, 小檗碱、巴马汀等重要生物碱也呈现较大的变化趋势, 茵芋碱、5-羟基小檗碱和四氢巴马汀等成分配伍二妙丸类方后消失. 研究显示, 基本方二妙丸中的黄柏生物碱是二妙丸类方中保护HUVEC起重要作用的物质基础, 其相对含量变化与配伍类方后的功效变化一致.     

靶向亲和-液相色谱-质谱联用技术快速筛选黄藤总生物碱中乙酰胆碱酯酶抑制剂

采用靶向亲和-液相色谱-质谱联用技术(Target molecule affinity-LC-ESI-MSn)快速筛选黄藤总生物碱中能够抑制乙酰胆碱酯酶活性的成分,共筛选出12种具有潜在抑制乙酰胆碱酯酶的活性成分,并鉴定了6种成分,分别为黄藤素(Palmatine)、小檗碱(Berberine)、药根碱(Jatrorrhizine)、巴马汀红碱(Palmatrubine)、7,8-二氢-8-羟基小檗碱(7,8-Dihydro-8-hydroxyberberine)、Groenlandicine,结合体外酶学实验对这6种化合物进行了活性验证实验.结果表明,黄藤素抑制活性最强,其抑制作用强于阳性对照药盐酸多奈哌齐,说明黄藤素具有开发成抗阿尔茨海默症药物的潜力.本方法简单、快速、准确地从复杂的中药提取物中筛选出具有抑制乙酰胆碱酯酶活性的成分,适用于复杂体系中的高通量筛选.     

基于Mn掺杂ZnS量子点室温磷光法检测盐酸巴马汀

本文以3-巯基丙酸(3-Mercaptopropionic Acid,MPA)为稳定剂,采用水相合成法制备了Mn掺杂ZnS量子点(Mn∶ZnS QDs),基于Mn∶ZnS QDs的室温磷光性质,盐酸巴马汀(Palmatine Hydrochloride,PaH)可与Mn∶ZnS QDs发生静电作用,使得Mn∶ZnS QDs发生室温磷光猝灭效应,从而发展了一种高效、快速检测人体体液中痕量PaH的新方法。实验结果表明,当PaH的浓度在0.75~30μmol/L范围时,其浓度与室温磷光猝灭强度(ΔIRTP)呈良好的线性关系,相关系数为0.996,检出限为0.35μmol/L,加标回收率为94.0%~103.3%。     

Photochemistry and photocytotoxicity of alkaloids from Goldenseal (Hydrastis canadensis L.) 3: effect on human lens and retinal pigment epithelial cells

The dried root or rhizome of Goldenseal (Hydrastis canadensis L.) contains several alkaloids including berberine, hydrastine, palmatine and lesser amounts of canadine and hydrastinine. Preparations derived from Goldenseal have been used to treat skin and eye ailments. Berberine, the major alkaloid in Goldenseal root powder, has been used in eye drops to treat trachoma, a disease characterized by keratoconjunctivitis. Berberine and palmatine are also present in extracts from Berberis amurensis Ruprecht (Berberidaceae) which are used to treat ocular disorders. We have previously shown that Goldenseal alkaloids are phototoxic to keratinocytes (Chem Res Toxicol. 14, 1529, 2001; ibid 19, 739, 2006) and now report their effect on human lens and retinal pigment epithelial cells. Human lens epithelial cells (HLE-B3) were severely damaged when incubated with berberine (25 microM) and exposed to UVA (5 J cm(-2)). Under the same conditions, palmatine was less phototoxic and hydrastine, canadine and hydrastinine were inactive. Moderate protection against berberine phototoxicity was afforded by the antioxidants ascorbate (2 mM) and N-acetylcysteine (5 mM). When exposed to UVA (5 J cm(-2)) both berberine (10 microM) and palmatine (10 microM) caused mild DNA damage as determined by the alkaline comet assay which measures single strand breaks. Berberine and palmatine are the only Goldenseal alkaloids with appreciable absorption above 400 nm. Because light at wavelengths below 400 nm is cut off by the anterior portion of the adult human eye only berberine and palmatine were tested for phototoxicity to human retinal pigment epithelial (hRPE) cells. Although berberine did damage hRPE cells when irradiated with visible light (lambda > 400 nm) approximately 10 times higher concentrations were required to produce the same amount of damage as seen in lens cells. Palmatine was not phototoxic to hRPE cells. Neither berberine nor palmatine photodamaged DNA in hRPE. Infusions of Goldenseal are estimated to contain approximately 1 mM berberine, while in tinctures the alkaloid concentration may be more than 10 times higher. Our findings show that eyewashes and lotions derived from Goldenseal or containing berberine must be used with caution when the eyes are exposed to bright sunlight but that oral preparations are not likely to cause ocular phototoxicity.     

In vitro, in situ and in vivo studies on the anticandidal activity of Cassia fistula seed extract

Cassia fistula seeds have many therapeutic uses in traditional medicine practice. The present investigation was undertaken to demonstrate the anticandidal activity of the C. fistula seed extract at ultra-structural level through transmission electron microscope (TEM) and scanning electron microscope (SEM) observations. The effect of seed extract on the growth profile of the Candida albicans was examined via time-kill assays and in vivo efficacy of the extract was tested in an animal model. In addition, the anticandidal effect of seed extract was further evaluated by microscopic observations using SEM and TEM to determine any major alterations in the ultrastructure of C. albicans. The complete inhibition of C. albicans growth was shown by C. fistula seed extract at 6.25 mg/mL concentration. The time-kill assay suggested that C. fistula seed extract had completely inhibited the growth of C. albicans and also exhibited prolonged anti-yeast activity. The SEM and TEM observations carried out to distinguish the metamorphosis in the morphology of control and C. fistula seed extract-treated C. albicans cells revealed the notable effect on the outer cell wall and cytoplasmic content of the C. albicans and complete collapse of yeast cell exposed to seed extract at concentration 6.25 mg/mL at 36 h. The in vitro time-kill study performed using the leaf extract at 1/2, 1 or 2 times of the MIC significantly inhibited the yeast growth with a noticeable drop in optical density (OD) of yeast culture, thus confirming the fungicidal effect of the extract on C. albicans. In addition, in vivo antifungal activity studies on candidiasis in mice showed a 6-fold decrease in C. albicans in kidneys and blood samples in the groups of animals treated with the extract (2.5 g/kg body weight). The results suggested that the C. fistula seed extract possessed good anticandidal activity and is a potential candidate for the development of anticandidal agents.     

LCST demixing in poly(vinyl methyl ether)/water studied by means of a High Resolution Ultrasonic Resonator

The inhibitory effects of three berberine alkaloids (BAs) from Coptis chinensis Franch on Bifidobacterium adolescentis growth were investigated by microcalorimetry. The growth rate constant (k) and maximum heat-output power (Pmax) decreased and peak time of maximum heat-output power (tp) prolonged with the increase of BAs concentration. Half inhibitory ratios (IC50) BAs were respectively 790.3 (berberine), 339.6 (coptisine) and 229.8 μL⁻¹ (palmatine), which indicated the sequence of their antimicrobial activity: berberine<coptisine<palmatine. Combined with previous findings, the sequence which could show the bioactivity of Bacillus shigae and Escherichia coli was: berberine>coptisine>palmatine. The structure-function relationship of BAs indicated that the functional group methylenedioxy or methoxyl at C2 and C3 might be the major group inducing the activities of BAs on E. coli and B. adolescentis. Meanwhile, the substituent groups at C2, C3, C9 and C10 almost had equal effect on B. shigae.     

COMPARATIVE STUDIES ON THE PHOTOSENSITIZING POTENCY OF 5-METHOXYPSORALEN and 8-METHOXYPSORALEN AS MEASURED BY CYTOLYSIS IN PARAMECIUM CAUDATUM AND TETRAHYMENA PYRIFORMIS, AND GROWTH INHIBITION AND SURVIVAL IN CANDIDA ALBICANS

Abstract— The photosensitizing potencies of 5-methoxypsoraIen and 8-methoxypsoralen were investigated using the following biological end-points for lethality: (i) cytolysis in the protozoans Paramecium caudatum and Tetrahymena pyriformis, (ii) inhibition of growth in the yeast Candida albicans and (iii) survival as measured by colony counts in Candida albicans. In all cases, 5-methoxypsoralen proved to be the more potent photosensitizing agent. The preliminary action spectra of the 2 compounds for growth inhibition in C. albicans were similar and showed maximal spectral efficiency in the 320–340 nm waveband. The molecular basis for the described end-points is unknown. Although it is well known that 5-methoxypsoralen and 8-methoxypsoralen readily photoreact with DNA, it is considered that photo-reaction with protein should also be given serious consideration as the possible lethal event. The superior effect of 5-methoxypsoralen is in accordance with some of the physical and photochemical properties of this molecule, but this result is at variance with other studies in different test systems that have been used to compare the photobiological efficacy of these 2 compounds.     

Microcalorimetric investigation of the antibacterial activity of curcumin on Staphylococcus aureus coupled with multivariate analysis

The antibacterial effect of Curcumin on Staphylococcus aureus growth was evaluated by microcalorimetry. The heat flow power?Ctime curves and nine quantitative parameters of the S. aureus growth were applied to investigate the inhibitory effect with Curcumin. By analyzing these curves and some quantitative parameters using multivariate analytical methods, similarity analysis and principal component analysis, the antibacterial activity of Curcumin on S. aureus could be accurately evaluated from the change of the two main parameters, the second exponential growth rate constant k ₂ and the maximum heat flow power P _m ² . The main two thermal parameters played more important role in the evaluation: at low concentration (0?C10.5???g?mL^?1), Curcumin hardly influence the growth of S. aureus, while at high concentration (10.5?C43.4???g?mL^?1) it could notably inhibit the growth. All these illustrated that the antibacterial activity of Curcumin on S. aureus was enhanced with the increase of the concentration of this compound. This study might provide an useful method and idea accurately evaluate the antibacterial effects of Curcumin, which provides some useful methods for evaluate the nature antibacterial agents.     

Influences of levofloxacin salts on the metabolism of Escherichia coli by microcalorimetry

A microcalorimetric technique was used to evaluate the influence of both Levofloxacin lactate in sodium chloride injection (drug A) and Levofloxacin hydrochloride in sodium chloride injection (drug B) on the metabolism of Escherichia coli. By means of an isothermal calorimeter and ampoule method at 37 °C, the power-time curves of E. coli growth were obtained under different conditions. The parameters such as the growth rate constant k, maximum power output P _m, time t _m corresponding to the maximum power output and inhibitory ratio I of these two drugs were obtained. The results reveal that the inhibitory abilities enhance with increasing concentrations of the two drugs. The critical growth concentration and the half-inhibitory concentration IC₅₀ were 0.15 and 0.079 μg mL^?1 (for drug A), 0.13 and 0.061 μg mL^?1 (for drug B), respectively. These results show the drug A has slightly better inhibitory effect on E. coli than that of drug B.