Applications of Molecular Imprinting Technology in the Study of Traditional Chinese Medicine

Traditional Chinese medicine (TCM) is one of the most internationally competitive industries. In the context of TCM modernization and internationalization, TCM-related research studies have entered a fast track of development. At the same time, research of TCM is also faced with challenges, such as matrix complexity, component diversity and low level of active components. As an interdisciplinary technology, molecular imprinting technology (MIT) has gained popularity in TCM study, owing to the produced molecularly imprinted polymers (MIPs) possessing the unique features of structure predictability, recognition specificity and application universality, as well as physical robustness, thermal stability, low cost and easy preparation. Herein, we comprehensively review the recent advances of MIT for TCM studies since 2017, focusing on two main aspects including extraction/separation and purification and detection of active components, and identification analysis of hazardous components. The fundamentals of MIT are briefly outlined and emerging preparation techniques for MIPs applied in TCM are highlighted, such as surface imprinting, nanoimprinting and multitemplate and multifunctional monomer imprinting. Then, applications of MIPs in common active components research including flavonoids, alkaloids, terpenoids, glycosides and polyphenols, etc. are respectively summarized, followed by screening and enantioseparation. Related identification detection of hazardous components from TCM itself, illegal addition, or pollution residues (e.g., heavy metals, pesticides) are discussed. Moreover, the applications of MIT in new formulation of TCM, chiral drug resolution and detection of growing environment are summarized. Finally, we propose some issues still to be solved and future research directions to be expected of MIT for TCM studies.     

双膦酸化合物对基质金属蛋白酶的抑制效应及机理

根据基质金属蛋白酶(MMPs)的活性位点附近三维空间结构, 设计合成了4种双膦酸类化合物, 利用酶促反应动力学方法对比测试了双膦酸化合物及阿伦磷酸钠(Alendronate)对MMPs的抑制效果; 结合分子对接方法以及荧光滴定光谱研究了双膦酸化合物与MMPs的分子识别和作用机理, 并得到了二者的结合模型.     

Expression Optimization and Characterization of the Catalytic Domain of Human MT3-MMP

Introduction Matrixmetalloproteinases(MMPs)areafamilyof calciumandzincrequiringendoproteinasesthattogether candegradeallthemaincomponentsoftheextra cellu larmatrixandbasementmembranes[1].MMPsarein volvedinawiderangeofproteolyticevents,innormal andpatholog…     

用MnO_4~-淬灭H_2QS-OP荧光体系测定中草药微量锰

本文报道使用非离子表面活性剂——聚乙二醇辛基苯基醚(OP乳化剂)对8-羟基喹啉-5-磺酸荧光体系的增敏增稳作用,进行中草药微量锰的淬灭测定,在最佳优选的实验条件下,重现性和灵敏度均有提高,取得了良好的效果。     

Network pharmacology-based approach of novel traditional Chinese medicine formula for treatment of acute skin inflammation in silico

Matrix metalloproteinase-9 (MMP-9) appears to play an important role in acute skin inflammation. Subantimicrobial dose of tetracycline has been demonstrated to inhibit the activity of MMP-9 protein. However, long-term use tetracycline will induce side effect. The catalytic site of MMP-9 is located at zinc-binding amino acids, His401, His405 and His411. We attempted to search novel medicine formula as MMP-9 inhibitors from traditional Chinese medicine (TCM) database by using in silico studies. We utilized high-throughput virtual screening to find which natural compounds could bind to the zinc-binding site. The quantitative structure-activity relationship (QSAR) models, which constructed by scaffold of MMP-9 inhibitors and its activities, were employed to predict the bio-activity of the natural compounds for MMP-9. The results showed that Celacinnine, Lobelanidine and Celallocinnine were qualified to interact with zinc-binding site and displayed well predictive activity. We found that celallocinnine was the best TCM compound for zinc binging sites of MMP-9 because the stable interactions were observed under dynamic condition. In addition, Celacinnine and Lobelanidine could interact with MMP-9 related protein that identified by drug-target interaction network analysis. Thus, we suggested the herbs Hypericum patulum, Sedum acre, and Tripterygium wilfordii that containing Celallocinnine, Celacinnine and Lobelanidine might be a novel medicine formula to avoid the side effect of tetracycline and increase the efficacy of treatment.     

高效液相色谱串联质谱法测定中药注射剂对P815细胞脱颗粒后上清液中组胺含量

建立了液相色谱-串联质谱(LC-MS/MS)检测P815细胞脱颗粒后上清液中组胺含量的方法。采用C18柱(4.6 mm×100 mm,3.5μm)进行分离,以体积比为60∶40的乙腈-0.5 mmol/L乙酸铵溶液(含0.3%甲酸)为流动相,流速0.4 mL/min;在电喷雾正离子化模式下,采用多离子反应检测方式(MRM)进行检测;检测离子对为m/z 112>95、112>68。在优化条件下,样品可在6 min内完成分析,细胞液中组胺在4.0～128.0μg/L范围内线性关系良好(r2=0.997 9)。定量下限(S/N>10)为1.2μg/L,检出限(S/N=3)为0.3μg/L。在加标水平为16、64、128μg/L时,细胞上清液中组胺的平均回收率为90%～94%,相对标准偏差(n=6)为4.1%～5.9%。该方法简便、灵敏、重复性好,可用于中药注射剂对P815细胞脱颗粒后上清液中组胺含量的测定。     

基因芯片用于组分中药新双龙方的配伍机制研究

采用基因芯片技术研究了中药新双龙方及其有效组分人参总皂苷和丹参总酚酸治疗急性心肌梗塞(AMI)大鼠的作用机制. 以结扎大鼠冠脉左前降支的方法制备急性心梗模型, 采用大鼠全基因组表达谱芯片分别筛选出AMI大鼠给药前及新双龙方与组分给药后的差异表达基因(Ratio>2或<0.5), 通过对4组差异基因的综合分析, 比较了新双龙方与组分作用方式的异同, 从分子调控水平上探讨复方配伍后的增效作用, 同时对与药效作用机理直接相关的钙离子转运相关的Cacna1d基因和血管修复相关的Flnb基因进行了实时荧光逆转录聚合酶链式反应(RT-PCR)定量. 结果表明, 新双龙方与2种有效组分对心梗大鼠基因表达的影响不同. 对差异基因的聚类分析以及药物的调控作用比较结果均表明复方比组分的疗效更好; 通路分析结果表明, 新双龙方与组分均主要通过调控钙信号转导通路、 丝裂原活化蛋白激酶信号转导通路(MAPK)信号等通路发挥药效作用, 且复方涉及更多通路; 筛选出了复方与组分共同调控的靶点基因, 其在3组中具有不完全相同的表达趋势, 但复方具有更积极的调控作用. 本研究结果从分子调控角度证实了复方新双龙方的配伍优势.     

QuEChERS-气相色谱法快速检测五十种中药材中九种有机氯农药残留的方法研究

建立了简便、快速测定五十种中药材中九种农药的多残留检测方法.以体积比95:4:1的乙腈-水-1%乙酸混合溶液作为提取剂,QuEChERS(Quick,Easy,Cheap,Effective,Rugged,Safe)方法进行净化,采用气相色谱法进行检测.结果表明:九种农药在1.0～100ng/mL范围内线性良好,3个添...     

基因芯片研究蟾酥急性毒性及配伍减毒机制

利用基因芯片技术研究蟾酥对大鼠心脏的急性毒性和其组方成麝香保心丸后的配伍减毒机制。通过表达谱芯片检测药物作用后的基因表达差异,对差异表达基因进行生物信息学研究并结合实时荧光定量PCR分析。结果表明低剂量蟾酥可以通过干扰离子稳态和肌动蛋白构建影响心脏的收缩,同时还会导致心脏细胞的抗凋亡和脂类代谢等应激反应;高剂量蟾酥除进一步干扰离子稳态和肌动蛋白构建外,还会引发铁离子蓄积,最终可能导致细胞凋亡;且蟾酥对心脏的影响具有剂量依赖性;蟾酥组方成麝香保心丸后,上述的影响均不明显,主要影响到血压调节和心肌修复等作用,体现了中药配伍的减毒作用。     

氢化物发生-原子荧光光谱法测定中草药中的痕量铋

本文报道了采用氢化物-原子荧光法测定中草药中的痕量铋。试验了酸介质和还原剂用量对测定铋的影响,考察了共存金属离子的干扰。在最佳测定条件下,方法检出限为0.1μg/L,线性范围为0.1~200μg/L,加标回收率为91.0%~104.2%。     

火焰原子吸收光谱法分析中药中锌的形态

采取正辛醇水分配体系模拟中药水煎液中锌在人体胃肠中的分配情况，用原子吸收光谱法测定了十二味中药药材及其水剪液中锌的含量，水煎液中水溶态锌、醇溶态锌的含量。实验结果表明，中药中锌的水煮溶出率较氏，中药中锌的形态与中药成分，配伍情况及作用靶位（胃肠）的酸度有关，酸度及配伍对中药中锌的溶出率及水煎液中锌的形态影响较大。     

聚类分析辅助中药寡糖电泳分析鉴定中药

基于中药多糖结构的复杂性和特征性,针对多糖部分降解后的寡糖片段,建立了一种采用毛细管区带电泳法(CZE)分离分析中药寡糖,并利用其特征性电泳谱图信息,结合聚类分析(CA)进行中药鉴定的方法。该方法以1-苯基-3-甲基-5-吡唑啉酮(PMP)为寡糖柱前衍生化试剂,对3个科属的6种中药如黄精、玉竹等同时进行鉴定。采用的电泳条件:未涂层熔融石英毛细管柱(49 cm(有效长度40 cm)×50 μm),以50 mmol/L磷酸盐缓冲液(pH 2.5)为运行缓冲液,检测波长为245 nm,运行电压为15 kV,虹吸进样10 cm×4 s,柱温为室温。结果表明聚类分析辅助中药寡糖电泳分析法可有效用于3个科属6种中药的鉴定。本方法结果可靠,重现性好,可以作为中药鉴定的一种有效手段。     

高分辨率电喷雾飞行时间质谱测定丹参脂溶性组分

丹参 [1] (Salvia miltiorrhiza Bage)为唇形科鼠尾草属多年生草本植物 ,临床上用于治疗妇科病、冠心病、缺血性中风及动脉粥样硬化等症 .近年来 ,飞行时间质谱 (TOF- MS)的分辨率优于 1 0 4 [2 ] ,尤其是再配合 ESI/MAL DI/APCI及新型脉冲电喷雾 [3 ]等离子源 ,使之成为当今最     

质谱技术在中药研究中的应用进展

中药是中华传统医学的瑰宝,在我国已广泛应用于疾病预防和治疗中,但其存在化学成分复杂、作用机制不明确等问题,制约了中药现代化及国际化的发展进程。质谱技术具有高灵敏度、高选择性、高稳定性及高通量的特点,特别适合于中药复杂成分及其代谢物的定性定量分析。该文综述了近年来质谱技术在中药成分鉴定及质量控制、中药代谢组学及中药药代动力学研究方面的典型应用,并对存在的问题及改进方法进行了展望。     

中药补气与活血分子作用机理的计算机模拟

采用分子相似性分析、分子对接和生物网络技术等计算机模拟方法对治疗冠心病的中药活血与补气的分子作用机理进行研究. 结果表明, 计算机模拟方法可以体现化合物的结构差异, 并提示中药中化学成分与相关靶标相互作用的分子机理, 而生物网络的构建和分析可以将化学成分的聚类、差异, 以及有效成分与相关靶标的复杂分子作用机理可视化.     

Role of counter-current chromatography in the modernisation of Chinese herbal medicines

This review focuses on the growing popularity of using counter-current chromatography (CCC), with its liquid stationary phase, as one of the prime methods for isolating compounds from Chinese herbal medicines (CHMs). 198 publications are reviewed covering 108 different plant species from 56 plant families. These describe the isolation of 354 different molecules across a wide range of polarities, chemical classes and molecular weights (in the range 100–1000 Da). The suitability of CCC for the separation of active compounds from CHM, the phase systems used, how CCC has developed in China, compounds isolated, CCC instrumentation, performance, operational issues and innovations, all supported by detailed cross-referencing, are described. It is concluded that CCC is making an increasingly important contribution to the modernisation of Chinese herbal medicines.     

中药指纹图谱研究的某些进展与展望

在中药现代化的过程中,中药的质量控制标准直接影响到中药的国际化问题。近年来,国内很多学者对中药指纹图谱用于质量控制的问题进行了深入的研究,取得了很多成果。本文着重就近几年来中药指纹图谱的某些进展进行了综述,内容包括气相色谱指纹图谱、液相色谱指纹图谱以及其他指纹图谱。最后就指纹图谱研究的发展提出了几点看法。共引用文献161篇。     

Association of Matrix Metalloproteinase-9 and p53 Gene Polymorphisms with Genetic Susceptibility to No-small-cell Lung Cancer

Matrix metalloproteinase-9(MMP-9) and p53 genes play an essential role in the multi-step process of tumorigenesis in lung cancer. Single nucleotide polymorphisms(SNPs) of MMP-9 and p53 genes are associated with the risk and progression of many cancers. In this study, we evaluated the association of the R279Q polymorphism of MMP-9 or the A1/A2 polymorphism of p53 gene with the risk of no-small-cell lung cancer(NSCLC) in Han population of Northeast China. We examined the frequency of SNPs in the two kinds of ...     

一种基于DAD二极管阵列检测器技术的中药指纹图谱方法

用HPLC/DAD分别建立血府逐瘀口服液单波长指纹图谱和最大吸收指纹图谱, 对比两者峰数量和面积的变化情况, 采用药典委员会2004A版相似度软件评价两种方法的差异. 结果表明, 最大吸收指纹图谱比单波长指纹图谱或多波长指纹图谱更能全面地反映血府逐瘀口服液的色谱峰信息, 并且相似度结果差异明显. 该方法信息量大, 中药的整体性、宏观性及复杂性分析相适应.