载紫杉醇的大黄酸偶联物纳米胶束的制备及初步评价

目的制备载紫杉醇的D-α-生育酚聚乙二醇1000琥珀酸酯(D-α-tocopheryl polyethylene glycol 1000 succinate,TPGS)修饰的羧甲基壳聚糖-大黄酸偶联物(PTX/TPGS-CR)纳米胶束,并对其进行初步评价。方法采用透析法,以载药量、包封率及粒径为指标,通过单因素考察优化PTX/TPGS-CR纳米胶束的制备工艺并进行验证。以溶血实验及血管刺激性实验初步考察PTX/TPGS-CR纳米胶束的安全性。四甲基偶氮唑盐微量酶反应比色法(MTT)法考察PTX/TPGS-CR纳米胶束对Hela细胞的毒性。通过激光扫描共聚焦显微镜定性和流式细胞仪定量考察Hela细胞对PTX/TPGS-CR纳米胶束的摄取情况。结果制备工艺优化后制得的PTX/TPGS-CR纳米胶束粒径为(197.3±4.4)nm,PDI为(0.131±0.021),电位为(-31.8±0.5)mV,载药量为(48.20±3.03)%,包封率为(87.26±4.91)%。溶血实验结果表明,其溶血率低于1.71%;血管静脉注射无明显刺激性。其对Hela细胞的杀伤作用具有浓度和时间依赖性,能被Hela细胞高效摄取。结论PTX/TPGS-CR纳米胶束载药量和包封率高,安全性好,其体外抗肿瘤活性稍优于Taxol?。     

大黄酸通过抑制CDK9/STAT3途径防治低氧性肺动脉高压

目的：探究中药单体大黄酸对低氧性肺动脉高压（HPH）的影响及其作用机制。方法：以SPF级SD大鼠为研究对象,设置常氧组（N）、低氧组（H）、低氧+大黄酸组（HR）。Western blot法检测每组大鼠体内提取的肺组织匀浆中p-CDK9、CDK9、p-STAT3/STAT3、PCNA、Bcl-2、Bax、Caspase-3、Cleaved-Caspase-3（CC-3）蛋白表达差异。以右心导管法插管检测平均肺动脉压（mPAP）评估大鼠的血流动力学情况,称重法检测右心肥厚程度,HE染色后观察血管形态以评估大鼠的肺血管重塑情况。结果：与N组比,H组肺匀浆中p-CDK9、CDK9、p-STAT3/STAT3、PCNA、Bcl-2 蛋白表达均上调,而Bax、Caspase-3、CC-3 蛋白表达下调（均P ＜0.05）,H组大鼠的mPAP及右心肥厚指数显著上升（均P ＜0.05）,管壁面积（WA）/管总面积（TA）明显增加（P ＜0.05）,提示大鼠的肺血管重塑程度明显加重;与H组比,HR组肺匀浆中p-CDK9、CDK9、p-STAT3/STAT3、PCNA、Bcl-2蛋白表达均下调,而Bax、Caspase-3、CC-3 蛋白表达上调（均P ＜0.05）,mPAP及右心肥厚指数显著降低（均P ＜0.05）,WA/TA明显降低（P ＜0.05）,提示肺血管重塑改善。结论：大黄酸可能通过抑制CDK9的表达及磷酸化,抑制STAT3的磷酸化,缓解低氧诱导的大鼠肺动脉高压,改善肺血管重塑和右心肥厚。     

基于支持向量机的5种大黄苷元治疗脑缺血的配伍研究

目的筛选大黄中5种能够治疗脑缺血的大黄苷元(芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚)的最佳配伍。方法采用均匀设计法将大鼠分为14组,每组15只。采用线栓法制备大鼠大脑中动脉闭塞(MCAO)模型,以神经功能症状评分、脑梗死面积为指标,探讨不同配伍的大黄苷元对脑缺血大鼠的影响,同时基于支持向量机(SVM)建立大黄苷元药效预测模型。结果经SVM回归分析,得到大黄苷元最优配伍为芦荟大黄素6.653 4 mg/kg、大黄酸26.000 8 mg/kg、大黄素11.004 2 mg/kg、大黄酚3.841 4 mg/kg和大黄素甲醚3.862 0 mg/kg。结论不同配比的大黄苷元能有效改善大鼠脑缺血的各个指标,采用均匀设计结合SVM的模拟预测方法优选出了5种大黄苷元组分的最佳配比。     

Anti‐fibrotic and Anti‐tumorigenic Effects of Rhein,a Natural Anthraquinone Derivative,in Mammalian Stellate and Carcinoma Cells

Anthraquinone compounds have been recognized to possess antiinflammatory, anti‐fibrotic and anti‐tumour properties and thus applied in human and veterinary therapeutics as active substances of medicinal products. Amongst the anthraquinones isolated from Rheum palmatum, also known as da‐huang, rhein was detected as one of the highest metabolite contents in the bloodstream of mammals. The biological activities of rhein therefore deserve detailed investigation. In this study, we aimed to delineate the mechanism of inhibitory actions of rhein on fibrotic and tumorigenic processes by means of various biochemical assays, such as immunofluorescent staining, real‐time polymerase chain reaction (PCR) and western blotting analyses in rat pancreatic stellate cells (LTC‐14), human pancreatic ductal adenocarcinoma cells (PANC‐1) and human colon carcinoma cells (SW480 and SW620). Our results demonstrated that the application of rhein notably suppressed the mRNA and protein levels of various fibrotic and tumorigenic mediators including alpha‐smooth muscle actin, type I collagen, fibronectin, N‐cadherin and matrix metalloproteinases in the testing mammalian cells. The mechanism of the suppressive actions of rhein was associated with the modulation of the sonic hedgehog and serine‐threonine kinase signalling pathways. In conclusion, we suggest that rhein may serve as a therapeutic or an adjuvant agent in anti‐fibrotic and anti‐tumorigenic approaches. Copyright © 2014 John Wiley & Sons, Ltd.     

赤芍、大黄药对提取物中丹皮酚和大黄酸的组织分布研究

目的:建立同时测定大鼠组织中丹皮酚和大黄酸含量的高效液相色谱法。方法:采用Phenomsil BDS C18(4.6 mm×250 mm,5μm),以乙腈-0.1%磷酸(37∶63)为流动相,流速1.0 m L·min-1,以橙黄决明素为内标,在274 nm下进行检测。结果:丹皮酚的线性范围为Y=-0.055+0.03X(r=0.993 2),日内和日间的RSD均4.1%;大黄酸的线性范围为Y=-0.152+0.014X(r=0.993 5),日内和日间的RSD均4.9%。组织样品的稳定性符合要求。结论:所建立的大鼠组织中丹皮酚和大黄酸HPLC测定方法,适用于大鼠体内赤芍、大黄药对的组织分布研究。     

A New Natural Compound with Cytotoxic Activity from Tripterygium hypoglaucum

Objective To find antitumor constituents from Tripterygium hypoglaucum. Methods The chloroform extract of T. hypoglaucum was separated by silica gel column chromatography and preparative HPLC. The structures of compounds isolated were identified by spectral analysis and chemical evidence. Results Seven compounds were isolated and identified as rhein ethyl ester (1), chrysophenol (2), physcion (3), emodin (4), wilfordine (5), wilforgine (6), and wilforine (7). The cytotoxic activities of the compounds against cancer cell lines were assayed. Conclusion Compound 1 is a new natural compound with strong activities against human cancer cell lines (A2780 and OVCAR-3). Compounds 2－4 are isolated from this genus plants for the first time. The possible structure-activity relationship among compounds 1－4 shows that the methoxy group or oxyethyl moiety might be responsible for the cytotoxity.     

高效液相色谱法同时测定黄氏响声丸中5种蒽醌类成分含量

目的:建立黄氏响声丸中5种蒽醌类活性成分的高效液相色谱含量测定方法,有利于制剂的质量控制.方法:采用高效液相色谱法,色谱柱:Shimadzu VP-ODS柱(4.6mm×250mm,5μm),流动相为乙腈-0.1%磷酸梯度洗脱,流速为1.0ml/min,检测波长为254nm,柱温30℃.结果:5种成分为芦荟大黄素0.0...     

大黄及其蒽醌对自身免疫性疾病治疗的研究进展

大黄始载于《神农本草经》,药理活性广泛。自身免疫性疾病是一类因机体免疫耐受功能受损而致免疫细胞及其成分对自身组织结构和功能破坏的疾病,包括动脉粥样硬化、多发性硬化、痛风、类风湿性关节炎、自身免疫性甲状腺炎、溃疡性结肠炎、1型糖尿病、IgA肾病等。近年临床和实验研究显示,大黄及其蒽醌类成分对自身免疫性疾病具有良好的治疗潜力和开发前景。该文以中医药理论为指导,综述了大黄及其主要活性成分蒽醌对自身免疫性疾病的治疗和干预作用并对其作用机制进行解析,还针对大黄及其蒽醌目前研究存在的问题提出见解,旨在为大黄及其蒽醌在自身免疫性疾病的临床治疗和科学研究方面提供参考。     

多元回归分析大黄不同炮制品中蒽醌与抑菌活性相关性

目的:采用多元回归分析不同大黄炮制品蒽醌含量和大黄抑菌能力之间的相互关系。方法:采用HPLC法建立大黄中蒽醌的指纹图谱,流动相为乙腈-0.1%磷酸溶液梯度洗脱,检测波长为254 nm。通过大黄不同炮制样品对大肠埃希菌、金黄色葡萄球菌、铜绿假单胞菌的抑制效果来评价其抑菌活性,采用主成分、多元线性回归分析大黄不同蒽醌与抑菌效力的关系。结果:建立大黄不同炮制品的HPLC指纹图谱;大黄蒽醌成分中芦荟大黄素和大黄酸在大黄抑菌中起主要作用,抑菌关联度均达到了90%以上。结论:大黄蒽醌成分与其抑菌效能存在显著相关性,其中芦荟大黄素和大黄酸起到主要作用。