少棘蜈蚣水提取物的抗菌活性

用含有大肠杆菌Escherichia coli K12D31、poly IC和昆虫生理盐水注射液诱导3~4 d的少棘蜈蚣Scolo-pendra subspinipes mutilans水提取物表现出较高的抗菌活性,经测定发现在设定的不同温度、pH值及离子强度条件下表现出良好的抗菌活性;对不同的蛋白酶表现出不同的敏感程度;除了对Proteus mirobilis未表现出抑菌外,对其它9种革兰氏阳性菌和阴性菌、真菌都有抗菌作用,表明有广谱抗菌活性;在600μg/m l以下,没有表现出溶血和凝集活性;电镜观察表明可损伤大肠杆菌的外壁,内容物外泄,菌体死亡。     

Alleviation of PEGylated Puerarin on Erythrocyte Hemolysis Induced by Puerarin in Glucose-6-phosphate Dehydrogenase- deficient Rats

Objective To explore and analyze the reducing hemolytic effects of PEGylated puerarin (PEG-PUE) on erythrocytes induced by PUE in glucose-6-phosphate dehydrogenase (G6PD)-deficient rats. Methods The rat model with G6PD-deficiency was established via sc injecting 1% acetylphenyl-hydrazine. Then the G6PD-deficient erythrocyte suspension obtained from this rat model was used to evaluate the hemolytic effects of PUE and the reducing hemolytic effects of PEG-PUE via hemolytic activity and erythrocyte osmotic fragility assay. Results It was found that PUE could cause a serious hemolysis to the erythrocyte suspension with the increase of drug concentration and the prolongation of drug incubation time, the hemolytic rate of PUE was up to 40%, while the addition of PEG-PUE to the erythrocyte suspension revealed no significant hemolysis. Additionally, the result of erythrocyte osmotic fragility indicated that PEG-PUE exerted a slight effect on the erythrocyte membranes, and the NaCl concentration that induced 50% hemolysis (32 mmol/L) was about one-third PUE. Conclusion These results demonstrate that PEG-PUE could play a significant role in reducing the side effect of hemolysis induced by PUE. The low hemolytic activity of PEG-PUE makes it a favorable candidate for in vivo tests and PEG-PUE could also provide the useful insight for the further formulation development as an innovative drug.     

人参皂苷溶血及抗溶血作用研究

目的：通过对几种单体人参皂苷溶血、抗溶血作用进行研究，得到人参皂苷浓度与溶血度间的关系曲线，探讨其作用规律。方法：不同浓度的单体人参皂苷溶液和相同浓度的兔血红细胞悬浊液组成的混合溶液充分作用后，用比色法测定溶血度；不同浓度的单体人参皂苷溶液与相同浓度的对照皂苷溶液混合后加入相同浓度的兔血红细胞悬浊液，三者组成的混合溶液充分作用后，用比色法测定溶血度并计算保护溶血百分数。通过数理统计软件对数据进行分析并拟合人参皂苷溶血或保护溶血作用的曲线。结果：在测定的人参皂苷各单体中分别或兼具有溶血和抗溶血的特性，且溶血和抗溶血能力各不相同。结论：原人参皂苷三醇型人参皂苷Re、Rg1、20（R）-Rg：、20（S）-Rg2和Rh。都具有抗溶血作用，其中20（R）-Rg2、20（S）-Rg2和Rh1，在较高浓度时还表现出溶血作用；原人参皂苷二醇型人参皂苷Rb1、Rb2、Rc和Rd均表现出抗溶血作用，其中Rd还在浓度较高时表现出了溶血作用。齐墩果酸型人参皂苷Ro在较低浓度便表现出抗溶血作用，未见溶血作用。     

Hemolytic polar steroidal constituents of the starfish Aphelasterias japonica

A reinvestigation of the polar steroid fraction from the starfish Aphelasterias japonica, collected near the Russian shore of the Sea of Japan, has afforded two new compounds, the disulfated quinovoside aphelasteroside C (1) and the monosulfated polyhydroxysteroid aphelaketotriol (2). Compounds 1 and 2 contain a unique 23-oxo-24-hydroxylated side chain that is unprecedented in marine steroids. The known compounds cheliferoside L1 (3), 3-O-sulfoasterone (4), forbeside E3 (5), and 3-O-sulfothornasterol A (6) were also isolated from this source. Compounds 1-3, 5, and 6 showed hemolytic activity to mouse erythrocytes.     

中药蜈蚣毒溶血活性试验

通过溶血试验对活体少棘蜈蚣,药材少棘蜈蚣和多棘蜈蚣的粗毒进行溶血活性比较。结果表明均有溶血活性,且药材蜈蚣毒活性较活体蜈蚣大大降低,陈药材蜈蚣毒活性较新鲜药材降低一倍,多棘蜈蚣毒活性明显高于少棘蜈蚣。     

Anticomplement activity of polyacetylenes from leaves of Dendropanax morbifera Leveille

The present study evaluated the anticomplement effect of polyacetylenes from Dendropanax morbifera (Araliaceae) in the classical pathway complement system. The leaves of D. morbifera were evaluated with regard to its anticomplement activity, and its active principles identified following activity‐guided isolation. An aqueous CCl₄ fraction of the leaves of D. morbifera exhibited significant anticomplement activity on the classical pathway complement system, which was expressed as total hemolytic activity. Three polyacetylenes isolated from the leaves of D. morbifera, namely (3S)‐falcarinol (1), (3S,8S)‐falcarindiol (2) and (3S)‐diynene (3). Compounds 1, 2 and 3 showed inhibitory activity against complement system with 50% inhibitory concentrations (IC₅₀) values of 87.3 µm , 15.2 µm and 39.8 µm . Among the compounds tested, 2 showed the most potent anticomplement activity (IC₅₀, 15.2 µm ). Copyright © 2010 John Wiley & Sons, Ltd.     

Studies on the antiinflammatory and related pharmacological activities of Psidium guajava: A preliminary report

The methanolic fraction of Psidium guajava fruit extract was found to possess significant inhibitory activity against carragenin, kaolin and turpentine-induced oedema formation. The fraction significantly inhibited protein exudation. The proliferative form of inflammation was significantly counteracted following cotton pellet-induced granuloma formation in rats. Acetic acid-induced algesia (writhing) in mice was significantly inhibited by the fraction. Significant antipyretic activity of the fraction was observed following yeast-induced pyrexia in rats. Potent antiarthritic activity was observed with the fraction against formaldehyde-induced chronic arthritis in rats.     

云芝多糖对小鼠免疫功能的影响

 云芝多糖２５０或５００ｍｇ·ｋｇ－１可显著提高正常小鼠经植物血球凝集素（ＰＨＡ）刺激的淋巴细胞转化率，血清抗绵羊红细胞（ＳＲＢＣ）血凝抗体效价，脾细胞产生溶血抗体能力，自然杀伤（ＮＫ）细胞及巨噬（Ｍ）细胞抗瘤活性。环磷酰胺（Ｃｙ）造成免疫抑制状态下，云芝多糖５００ｍｇ·ｋｇ－１可使淋巴细胞转化率、血清血凝抗体效价及脾细胞产生溶血素能力提高至接近正常水平。这些结果表明，云芝多糖对小鼠免疫功能有明显的增强作用。     

七叶一枝花和滇重楼提取物的制备、表征及其体外溶血作用分析

目的:比较七叶一枝花和滇重楼提取物体外溶血作用强弱,探讨不同结构类型重楼皂苷的溶血作用特点。方法:采用75%乙醇对七叶一枝花和滇重楼根茎进行提取,采用大孔吸附树脂对各类物质进行富集,采用快速制备色谱技术对目标组分群进行定向制备,基于对照品对照和在线紫外吸收光谱对总取物、部位和组分进行化学信息表征,采用分光光度法测定各样品的溶血率。结果:根据质量浓度-溶血率曲线计算七叶一枝花和滇重楼总提物的50%溶血浓度分别为34.97,46.52 mg·L~(-1)。在2.5~10 mg·L~(-1)时,七叶一枝花部位中仅有70%乙醇洗脱部位(PPC-D)显示了较强溶血作用,滇重楼部位中70%乙醇和95%乙醇洗脱部位(PPY-D和PPY-E)均显示溶血作用。5.0,10 mg·L~(-1)时PPC-D的溶血率分别为78.00%和95.68%,PPY-D(PPY-E)的溶血率分别为31.86%和63.29%(15.82%和55.25%)。在2.5~5.0 mg·L~(-1)时,七叶一枝花的组分呋甾皂苷和薯蓣皂苷未显示溶血作用,偏诺皂苷组分在质量浓度为2.5,5.0 mg·L~(-1)的溶血率分别为5.89%和90.33%;滇重楼的呋甾皂苷组分同样未显示溶血作用,偏诺皂苷(薯蓣皂苷)组分在质量浓度为2.5,5.0 mg·L~(-1)的溶血率分别为0.51%和57.75%(4.29%和49.39%)。结论:七叶一枝花和滇重楼均具有溶血作用,其中七叶一枝花强于滇重楼;七叶一枝花溶血成分群主要为偏诺皂苷类,滇重楼溶血成分群为偏诺皂苷和薯蓣皂苷;两者呋甾皂苷均无溶血作用;总提物溶血作用强弱主要由其中所含偏诺皂苷和薯蓣皂苷类成分的绝对含量和相对比例来决定。     

Antimicrobial investigation of semipurified fractions of Ginkgo biloba leaves

A total methanolic extract of Ginkgo biloba leaves was fractionated by solvent partition using ethyl acetate (fraction A), n-butanol (fraction B) and water (fraction C). The antimicrobial activity of the three fractions was evaluated using a number of Gram-positive and -negative bacteria and yeasts. The apolar fraction A appeared to be the most interesting because of its activity against several microorganisms; this fraction was further separated by high performance liquid chromatography, and shown to contain substances with strong inhibitory activity against Enterococcus faecalis 31, different from the major known chemical components of G. biloba leaves.     

Prediagnostic methods for the hemolysis of herbal medicine injection

Ethnopharmacological relevance

The Xue-Sai-Tong injection, a traditional Chinese medicine injection with total saponins extracted from Sanchi Ginseng, has been used for more than half a hundred years to treat coronary artery disease. The study is to establish a prediagnostic method for the hemolytic adverse effect of herbal medicine injection by taking Xue-Sai-Tong injection as an example.

Materials and methods

A new method named “fuzzy dissemination” was established to identify the hemolytic ginsenosides in Xue-Sai-Tong injection on the basis of fuzzy changes of individual ginsenosides in the injections altered by re-adding the fractions prepared from the total saponins and statistic analysis between hemolytic degrees and individual ginsenosides. Related substances test, safety tests and fingerprints of the injections in different batches were tested.

Results

HD₅₀, P₅₀ and interactions on hemolysis of individual ginsenosides were examined. Experiment indicated that the content of Rg₁, Rg₂, M₅₁ (an unknown ingredient with retention time at 51 min in HPLC) and M₇₀ in Xue-Sai-Tong injection showed a significant positive correlation with hemolytic degree, and the content of R₁, Re, Rb₁ and Rd showed a significant negative correlation with hemolytic activity. Furthermore HD₅₀ of injection exhibits superiority to other tests for the hemolysis of injections. Abnormal hemolysis in some batches of injections was observed, but there were no significant differences among injections of different batches in related substances test, safety test and fingerprints.

Conclusions

This is an original method to analyze active ingredients of a complicated integrity instead of studying on individual ingredients, it showed that the interactions of some individual ginsenosides and some unknown micro-ingredients in Xue-Sai-Tong injection were the major factors causing hemolysis, and this method could also be utilized in research of corresponding aspects. HD₅₀ of injection can reflect the changes of hemolytic property of injections caused by not only the change of active constituents of injection, but also the auxiliary materials. Thus it was recommended as an index for the hemolytic prediagnosis of the injections in practice.     

Anti‐complement activity of isolated compounds from the roots of Clerodendrum bungei Steud.

To determine the anti‐complement activity of natural diterpenes, chromatographic separation of the acetone‐soluble fraction from the roots of Clerodendrum bungei (Verbenaceae) led to the isolation of five diterpenoids. An acetone‐soluble extract of the roots of C. bungei exhibited significant anti‐complement activity on the classical pathway complement system, which was expressed as total hemolytic activity. Five compounds isolated from the roots of C. bungei, namely 12‐O‐β‐d ‐glucopyranosyl‐3,11,16‐trihydroxyabieta‐8,11,13‐triene (1), 3,12‐O‐β‐d ‐diglucopyranosyl‐11,16‐dihydroxyabieta‐8,11,13‐triene (2), ajugaside A (3), uncinatone (4) and 19‐hydroxyteuvincenone F (5). Compounds 1, 2, 3, 4 and 5 showed inhibitory activity against complement system with 50% inhibitory concentrations (IC₅₀) values of 24 µm , 138 µm , 116 µm , 87 µm and 232 µm . Among the compounds tested, 1 showed the most potent anti‐complement activity (IC₅₀, 24 µm ). Copyright © 2010 John Wiley & Sons, Ltd.     

桃红四物汤各分离部位对体外ADP诱导血小板聚集、凝血酶活性的影响及效应物质基础研究

评价桃红四物汤各分离部位对体外二磷酸腺苷(ADP)诱导血小板聚集及凝血酶活性的影响,探讨其效应物质基础。采用比浊法测定体外ADP诱导家兔血小板聚集率;采用改进凝血酶时间法测定抑制凝血酶活性;采用UPLC-Q-TOF-MS对各分离部位进行定性分析鉴定。桃红四物汤醇沉沉淀部位、醇沉上清液部位及20%~30%醇洗脱部位对ADP诱导的血小板聚集具有显著的抑制作用;水不溶部位、醇沉上清液部位及40%~70%醇洗脱部位具有显著的抑制凝血酶活性的作用;芳香酸类成分、苷类成分和苯酞类成分是醇沉沉淀部位、醇沉上清液部位及20%~40%醇洗脱部位的主要成分。桃红四物汤抑制ADP诱导家兔血小板聚集及抑制凝血酶活性的主要效应物质基础是芳香酸类成分、苷类成分和苯酞类成分。该实验为进一步探讨活血化瘀方桃红四物汤治疗血瘀证的物质基础及作用机制提供了科学依据。     

Cytotoxic triterpenoid saponins acylated with monoterpenic acid from Pithecellobium lucidum

Three new oleanane-type triterpene saponins, named pithelucosides A-C (1-3), together with two known saponins (4, 5) were isolated from the roots of Pithecellobium lucidum. The structures of the new saponins were established on the basis of extensive 1D and 2D NMR experiments and mass spectrometry and confirmed by acid and alkaline hydrolysis. Compounds 1-5 and 7 (pro-sapogenin obtained from the mild alkaline hydrolysate of 1) were evaluated for cytotoxic activity on five human tumoral cell lines (HCT-8, Bel-7402, BGC-823, A549, and A2780) and for hemolytic property against rabbit erythrocytes. Compounds 2-5 showed significant cytotoxic activities with IC50 values of 0.61-7.56 microM. All tested compounds did not exhibit any hemolytic activity in the concentration range 0.01-100 microM.     

A Kunitz proteinase inhibitor from corms of Xanthosoma blandum with bactericidal activity

Bacterial infections directly affect the world's population, and this situation has been aggravated by indiscriminate use of antimicrobial agents, which can generate resistant microorganisms. In this report, an initial screening of proteins with antibacterial activity from corms of 15 species of the Xanthosoma genus was conducted. Since Xanthosoma blandum corms showed enhanced activity toward bacteria, a novel protein with bactericidal activity was isolated from this particular species. Edman degradation was used for protein N-termini determination; the primary structure showed similarities with Kunitz inhibitors, and this protein was named Xb-KTI. This protein was further challenged against serine proteinases from different sources, showing clear inhibitory activities. Otherwise, no hemolytic activity was observed for Xb-KTI. The results demonstrate the biotechnological potential of Xb-KTI, the first proteinase inhibitor with antimicrobial activity described in the Xanthosoma genus.     

晋产远志种质资源溶血性研究

目的：研究晋产远志种质资源溶血特性，借以间接反映其品质状况。方法：采用溶血试验测定红细胞半数致死量（LD50）。结果：山西不同气候区远志资源具有明显差异。结论：晋产远志种质资源溶血特性与该资源分布的农业气候区具有一定相关性。     

黄黑小斑蝥纤溶活性蛋白分离纯化及性质研究

目的以药用斑蝥(Mylabris)为原料,从其中分离纯化得到一种具有纤溶性的蛋白MFP(Fibrinolyric Protein of Myla-bris)。方法采用硫酸铵分段盐分、透析和DEAE-32纤维素离子交换层析等纯化方法进行分离纯化,SDS-PAGE电泳测定其分子量,纤维蛋白平板法测定其纤溶活性。结果该目的蛋白相对分子质量约为95.5 kD,其水解纤维蛋白的相对比活力为14.7 u/mg;该成分在35℃下最稳定,65℃及以上活性丧失,金属离子K+,Na+对其活力无影响,Mn2+,Ca2+对其有明显抑制作用,最适pH为6.0。结论研究分离出的斑蝥活性蛋白确为一种具有纤溶性的蛋白组分,并证明其纤溶作用机理为纤溶酶原激活;体外溶栓实验表明,其对血栓的溶解具有一定作用;溶血实验结果表明,该纤溶活性蛋白溶液无溶血反应。     

226例新生儿溶血病ABO血型不合的检测与分析研究

目的：探讨由母婴ABO血型不合引起的新生儿溶血病（HDN）的血型分布及其溶血三项检测试验的重要性.方法：对226例临床新生儿溶血病进行ABO新生儿溶血（HDN）血清学检测,包括直接抗人球蛋白试验、红细胞抗体释放试验、血清游离抗体试验.结果：在226例临床新生儿溶血病中由ABO血型抗原不合引起的新生儿溶血病138例,占送检标本的61.06%,其抗体释放试验均为阳性,A型患儿为63例,阳性率45.65%,B型患儿为75例,阳性率54.35%.结论：ABO血型不合引起的新生儿溶血病检测以抗体释放试验阳性为确诊试验,ABO新生儿溶血病的明确诊断可为临床诊疗及预防提供可靠依据.     

金樱根提取物不同极性部位的体外抗氧化活性考察及其化学成分分析

目的:考察金樱根75%乙醇提取物及其不同极性萃取部位的抗氧化活性和化学组成。方法:采用有机溶剂萃取法将金樱根75%乙醇提取物分为二氯甲烷、乙酸乙酯、正丁醇和水部位,利用1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除试验、铁离子还原能力(FRAP)试验考察金樱根乙醇提取物及其不同极性萃取部位的体外抗氧化活性。采用紫外分光光度法测定各部位中总三萜、总酚、总鞣质和缩合鞣质的含量。运用SPSS24.0对各部位抗氧化活性与其化学成分含量进行Pearson相关分析,筛选活性部位及主要的活性成分;采用超高效液相色谱-四极杆飞行时间质谱法(UPLC-Q-TOF-MS/MS)对该活性部位化学成分进行分析,推测主要色谱峰的结构。结果:金樱根乙醇提取物及其不同极性萃取部位均有一定的抗氧化活性,且在一定浓度范围内呈显著剂量-效应关系,但不同部位的抗氧化活性有差异;在DPPH和ABTS自由基清除试验中,阳性药维生素C(VC)和各部位抗氧化能力排序为乙酸乙酯部位>VC>正丁醇部位>乙醇提取物>水部位>二氯甲烷部位;FRAP试验中乙酸乙酯部位的活性弱于VC,其他次序不变。含量测定结果显示,乙酸乙酯部位中总三萜、总酚、总鞣质和缩合鞣质的质量分数分别为3.81%,50.33%,3.32%,39.79%,正丁醇部位中依次为0.88%,41.42%,2.25%,23.55%,乙醇提取物中依次为2.90%,41.95%,3.43%,20.14%,水部位中依次为0,26.80%,16.90%,7.57%,二氯甲烷部位中依次为21.23%,12.90%,1.59%,6.17%。相关性分析结果显示金樱根中总酚和缩合鞣质含量与抗氧化活性呈正相关,总三萜含量与抗氧化活性呈负相关,总鞣质含量与抗氧化活性相关性不明显。经液质联用分析,从乙酸乙酯部位中共鉴定出26个化合物,包括11个缩合鞣质,4个可水解鞣质,6个三萜类,3个黄酮类,1个苯甲酸衍生物和1个绿原酸类化合物。结论:乙酸乙酯部位是金樱根发挥抗氧化作用的主要活性部位,而以缩合鞣质为主的酚类物质是其主要活性成分,可为天然抗氧化剂的开发提供实验依据。     

Two new biologically active triterpenoidal saponins acylated with salicylic acid from Albizia adianthifolia

Two new oleanane-type triterpene saponins, adianthifoliosides A (1) and B (2), were isolated from a 95% ethanolic extract of roots of Albizia adianthifolia. Their structures were elucidated mainly by using a combination of 600 MHz 1D and 2D NMR techniques (COSY, NOESY, TOCSY, HSQC, and HMBC) and by FABMS and HRESIMS. Compounds 1 and 2 were characterized as glycosides of acacic acid acylated by an o-hydroxybenzoyl unit. The crude saponin mixture (CSM), compounds 1 and 2 together with 3 and 4 (prosapogenins obtained from the mild alkaline hydrolysate of the CSM), were evaluated for immunomodulatory activity on the Jurkat T cell line and for hemolytic property against sheep erythrocytes. Compound 2 and, to a lesser extent, 1 and 3 were found to exhibit a dose-dependent immunomodulatory effect in the concentration range 10(-2)-10 microM, whereas 4 showed a lymphoproliferative activity in the same concentration range. Among the compounds tested, only 1 and 2 were found to be hemolytic.