HPLC法测定复方板蓝根胶囊中靛蓝、靛玉红的含量

目的:建立以高效液相色谱法测定复方板蓝根胶囊中靛蓝、靛玉红含量的方法。方法:色谱柱为SHLM-PACK-ODS C18(150mm×4.6mm,10μm),流动相为甲醇-0.1%醋酸铵-醋酸(70:30:1),流速为1.0mL.min-1,检测波长为286nm。结果:靛蓝、靛玉红进样量分别在0.0152～0.3048μg(r=0.9997)、0.0166～0.3312μg(r=0.9995)范围内与各自峰面积积分值呈良好线性关系;二者平均加样回收率分别为97.8%和96.6%,RSD分别为1.87%(n=6)和2.32%(n=6)。结论:本法简便、快捷、分离度高、重现性好,可用于复方板蓝根胶囊的质量控制.     

高效液相色谱法测定抗感利咽糖浆中靛玉红的含量

目的:建立抗感利咽糖浆中靛玉红的含量测定方法。方法:以Dikma DiamonsilTM C18(200 mm×4.6 mm,5μm)为色谱柱;流动相为甲醇-0.1%醋酸铵-醋酸(70:30:1);流速1 ml·min-1;检测波长289 nm;柱温40℃。结果:靛玉红在0.02～0.24μg范围内线性良好,r=0.999 5。平均加样回收率为101.3%,RSD 1.1%(n=6)。结论:该法操作简便,结果准确,重现性好,可作为该制剂的质量控制方法。     

青黛中靛玉红含量测定方法的研究

目的建立青黛中靛玉红含量的测定方法,为2005年版《中国药典》提供含量测定控制方法。方法采用薄层扫描法对青黛中靛玉红进行含量测定,以苯-氯仿-丙酮(5∶4∶1)为展开剂,λS=540nm,λR=700nm;同时,采用薄层色谱法对青黛中的靛蓝和靛玉红进行定性鉴别。结果靛玉红的回归方程为y=11.915x 1052.4,r=0.9967;平均回收率为99.40%,(n=8),RSD为3.50%。测得7批青黛样品,含量在0.161%~0.225%之间。结论该方法具有简便、迅速、灵敏、重现性好等特点,可用于控制青黛药材的质量。     

RP-HPLC测定大鼠静脉注射靛玉红后的血药浓度

目的 建立大鼠血浆中靛玉红浓度的测定方法.方法 采用HPLC测定,血浆样品用丙酮沉淀蛋白后直接进样,色谱柱为Diamonsil C18(150 mm×4.6 mm,5μm),流动相为甲醇-0.1 mol·L-1醋酸铵-冰醋酸(73:26:1),流速1.0 ml·min-1,检测波长287 nm.结果 靛玉红在0.025～1.600 μg·ml-1范围内线性关系良好(r=0.9997,n=7),高、中、低浓度测定的方法回收率为94.5%～105.0%,日内RSD为0.14%～5.46%(n=5),日间RSD为2.54%～6.27%(n=5).结论 所用方法预处理简便,专属性强、灵敏度高,适用于靛玉红未知样品的血药浓度测定.     

板蓝根中的抗病毒活性成分

目的：寻找板蓝根中的主要抗病毒成分。方法：利用色谱法和结晶法对成分进行分离，鸡胚法为抗病毒活性检验法。结果：分离得到3个化合物，2，4（1H，3H）喹唑二酮（Ⅰ）、表告依春（Ⅱ）、靛玉红（Ⅲ），其中Ⅱ显示出较强抗病毒活性？结论：表告依春为板蓝根抗流感病毒的主要有效成分之一。     

7-Bromoindirubin-3'-oxime uncovers a serine protease-mediated paradigm of necrotic cell death

The new 7-bromoindirubin-3'-oxime (7BIO) compound induces caspase-independent cell death in all cell lines tested to date. Irrespective of the cell line, a 25 microM treatment for 24 h is lethal for the entire cell population. In SH-SY5Y and Jurkat cells, 7BIO (25 microM) was found to collapse the mitochondrial transmembrane potential (DeltaPsi m) at only 2-3 h of treatment. Concomitantly mitochondria swelled, cristae disrupted and, after 9 h, external cell membranes ruptured. In addition, endoplasmic reticulum dilated and, unexpectedly, the acute cytoplasmic destruction yielded isolated nuclei with preserved morphology and DNA integrity. Furthermore, the process was independent of both Bax and Bak, since cell viability and DeltaPsi m decayed indistinguishably in double Bax-/-Bak-/- mouse embryonic fibroblasts (MEFs) and their wild type counterparts. Pharmacological inhibition of the mitochondrial permeability transition pore (MPTP) did not prevent 7BIO-induced DeltaPsi m loss in none of the aforementioned cell lines. Caspase-independent inducers of cell death like AIF (Apoptosis Inducing Factor), cathepsins and calpains were not involved. Only the chemical inhibitors of serine proteases and, particularly, AEBSF afforded a significant protection thus suggesting a process regulated by this type of enzymes. As far as we know, these features are quite unique once taken together. Therefore, we propose 7BIO is triggering a specific type of necrotic cell death. Finally, the cytotoxicity of 7BIO on apoptosis-resistant cells like double Bax-/-Bak-/- MEFs seems of great interest envisaging cancer therapy.     

大青叶和板蓝根药材及其制剂质量控制的研究

首次提出以2,4(1H,3H)喹唑二酮[2,4(1H,3H)quinazolinedione]作为马蓝、蓼蓝药材及其制剂的质量控制指标。此成分系最近发现的,含量稳定且有代表性,尤其适用于评价此类中药制剂的真伪优劣,为当前难以解决的感冒退热冲剂和板蓝根冲剂的质量监控提供了可靠的方法。特别要指出的是本文是在高效液相色谱法中采用变换检测波长和梯度洗脱的方法,一次进样即可完成三种化学性质不同组分的含量分析,缩短了分析时间,提高了分析效率。     

Indirubin shows anti-angiogenic activity in an in vivo zebrafish model and an in vitro HUVEC model

Aim of the study

Indirubin is an active ingredient of the traditional Chinese medicine, Dang Gui Long Hui Wan, commonly used for the treatment of chronic myelocytic leukemia (CML) and other inflammatory conditions. These anti-leukemic and anti-inflammatory activities may be mediated by anti-angiogenic action. To investigate the anti-angiogenic activity of indirubin, we tested its inhibitory effect on blood vessel formation in zebrafish embryos and on endothelial cell proliferation in culture.

Materials and methods

The anti-angiogenic activity of indirubin was tested using transgenic zebrafish embryos with fluorescent vasculature and human umbilical vein endothelial cells (HUVECs). Apoptosis was analyzed with a terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay.

Results

Indirubin dose-dependently inhibited intersegmental vessel formation in zebrafish embryos. It also inhibited HUVEC proliferation by the induction of cellular apoptosis and cell-cycle arrest at the G0/G1 phase.

Conclusions

The anti-angiogenic activity of indirubin may partly contribute to its anti-leukemic and anti-psoriatic properties and may be valuable for the treatment of diseases with excessive angiogenesis. The zebrafish model of angiogenesis was further validated in this study.     

靛玉红类衍生物电子结构与抗癌活性关系的研究

目的:研究靛玉红类衍生物电子结构与抗癌活性的构效关系。方法:采用CNDO/2分子轨道法计算靛玉红类衍生物分子的电子结构,应用分子力学方法计算分子的构象,并结合相关分析和逐步回归分析方法探讨其构效关系。结果:靛玉红类衍生物分子中4号氧原子的净电荷Q4、3号C原子的净电荷Q3与其抗癌活性显著相关,并得到与Q4的定量构效关系:Iw=-40.03-246.96×Q4。当分子的键角L1为125°-132°,扭转角L2为11°-12°时,分子活性构象能够与靶标的活性部位更好地结合。结论:靛玉红类衍生物分子构象中有两个活性点(即药效基团),这两个活性点提供电子给靶标的活性部位。而有利于它们之间结合的构象,都可使其相互间的结合力提高,从而使抗癌活性增加。     

高效液相色谱法测定自制板蓝根免煎颗粒中靛玉红含量

目的建立自制板蓝根免煎颗粒中靛玉红含量测定方法。方法用高效液相色谱法,流动相:甲醇∶氯仿(85∶15)(加0.1%三乙氨,醋酸调至中性),流速:1ml/min,检测波长:290nm,柱温:25℃。结果方法学考察合格,准确测定出自制板蓝根免煎颗粒中靛玉红含量。结论本法快速准确,可行性高。