Effects of Angong Niuhuang Pills against heart failure based on cross-scale polypharmacological study / 中国中药杂志

Angong Niuhuang Pills(AGNHP) are effective in clearing heat, removing the toxin, and eliminating phlegm for resuscitation. Clinically, it is widely used to treat various diseases such as febrile convulsion due to heat attacking pericardium, but its therapeutic effects on heart failure(HF) have not been well recognized. In this study, the profiles of differential metabolites regulated by AGNHP were identified by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). The underlying mechanism of AGNHP against HF was illustrated based on the integrated analysis of pharmacological data and metabolic molecular network. The HF model was induced by isoproterenol in mice. After oral administration of AGNHP for one week, cardiac functions in HF mice were evaluated by echocardiography, and serum samples of mice were collected for metabolomics analysis. Eight differential metabolites of AGNHP against HF were screened out through partial least square discriminant analysis(PLS-DA) and input into MetaboAnalyst for the analysis of metabolic pathways. Moreover, the critical metabolic pathways regulated by AGNHP were enriched according to the potential targets of major compounds in AGNHP. After AGNHP treatment, the recovered index of relative content of some metabolites underwent cross-scale fusion analysis with therapeutic efficacy data, followed by "compound-reaction-enzyme-gene" network analysis. It is inferred that the anti-HF effects of AGNHP may be attributed to the metabolism of arachidonic acid, amino acid, glycerophospholipid, and linoleic acid. The cross-scale polypharmacological analysis method developed in this study provides a new method to interpret scientific principles of AGNHP against HF with modern technologies.

Network pharmacology study of mechanism on xuesaitong injection against retinal vein occlusion / 中国中药杂志

Retinal vein occlusion (RVO) is a common clinical disease causing vision loss. Risk factors such as diabetes, atherosclerosis are closely associated with RVO. Xuesaitong injection is used extensively in clinical treatment of RVO, however the mechanism is still unclear. In this study, we investigated the protective effect of Xuesaitong injection on RVO rat model. Using a compound-target network of Xuesaitong on anti-RVO constructed by literature mining, we aim to elucidate the multi-compound, multi-target effect of Xuesaitong injection. Fifteen potential targets of Xuesaitong injection associated with inflammation, angiogenesis, apoptosis, and coagulation were identified in this study. VEGF, IL-1beta and IL-6, three important targets in the compound-target network were further experimentally validated. This study provided experimental evidence for Xuesaitong injection being effective in treating RVO and a network view on its anti-RVO mode of action through a multi-compound and multiple-target mechanism.

Study of screening nephroprotective bioactive substances based on triple-color fluorescence probes in Carthami flos / 中国中药杂志

In this study, an approach based on triple-color fluorescence probes was developed for screening potential nephro-protective bioactive substances. Three fluorescent probes (i. e. FDA, MTR and Hoechst 33342) were used to label HK-2 cells injured by doxorubicin hydrochloride, and cellular fluorescence images were subsequently acquired and analyzed by a cellular-fluorescence image microscopy platform. The established method was applied to screening 53 components of Carthami Flos, and three components C17, C18 and C19 were found to exhibit nephroprotective effects against doxorubicin hydrochloride induced injury on HK-2 cells. Eight compounds (i. e. hydroxysafflor yellow A, 6-hydroxykaempferol-3-O-rutinoside-6-O-glucoside, 6-hydroxykaempferol-3,6-di-O-gluco-side or 6-hydroxykaempferol-6, 7-di-O-glucoside, 6-hydroxykaempferol-3-O-rutinoside, 6-hydroxykaempferol-3-O-glucoside or 6-hydroxykaempferol-7-O-glucoside, rutin, isoquercetin, and kaempferol-3-O-rutinoside) in components C17, C18 and C19 were preliminarily identified by liquid chromatography-mass spectrometry (LC-MS). Isoquercetin, rutin, kaempferol-3-O-rutinoside, and hydroxysafflor yellow A were confirmed by comparing with reference substances, Further study indicated that these four compounds had moderate nephroprotective effects, while isoquercetin showed a significant nephroprotective effect in a dose-dependent manner. These results suggest that isoquercetin, rutin, kaempferol-3-O-rutinoside and hydroxysafflor yellow A might be the nephroprotective bioactive substances in Carthami Flos.

Strategies and key technologies for risk control and management in quality of Chinese medicine injections based on integrated pharmacology / 中国中药杂志

This paper focuses on the quality risk control and management of Chinese medicine (CM) injections. The most important technological requirements are analyzed, and a strategy for integrated pharmacology to study CM mechanism is proposed. A key technology system for quality risk control and management was further constructed. The strategy and technology system was finally applied to Shengmai injection for quality risk control and management.

Study on screening cardioprotective components from Zhishi Xiebai Guizhi Tang / 中国中药杂志

Zhishi Xiebai Guizhi Tang (ZSXBGZT), a famous traditional Chinese formula from Synopsis of Golden Chamber, composing of five herbal medicines, has been routinely used for the therapy of cardiovascular diseases in clinical. However, the active components of ZSXBGZT have not been investigated thoroughly. A reliable cell model was established by H2O2-induced injury in neonatal rat cardiomyocytes (NRCMs), with fluorescein diacetate (FDA) labelling the live cells, to screen cardioprotective substances. Seven components were found to have obvious cardioprotective effects through screening ZSXBGZT, Vc as the positive control. Eleven compounds were identified by liquid chromatography-mass spectrometry (LC-MS) from the active components C18, D14, D15, D16 and E09, and 6 of them were investigated further, which showed that hesperidin, neohesperidin and eriodictyol could protect NRCMs from H2O2-induced injury. In particular, eriodictyol showed significant cardioprotective effects in a dose-response relationship.

Development of a rapid screening method for discovering neuroprotective components from traditional Chinese medicine / 中国中药杂志

This study developed a method for rapid screening neuroprotective compounds with FDA (fluorescein diacetate) labeled SH-SYSY cells, which was injuried by Glu (L-glutamic acid). The cell viability was determined by fluorescence automatic mi-croscopy screening system. Then, neuroprotective components from Gegen Qinlian decoction were screened by the proposed method. The results demonstrated that 4 chemical components, C15, D06, D07 and E05 from Gegen Qinlian decoction, showed significantly neuroprotective effects. Furthermore, 8 compounds (i. e. daidzin, 3'-methoxydaidzin, liquiritin apioside, 6-C-L-alpha-arabinopyranosyl-8-C-beta-D-glucopyranosyl chrysin, isoliquiritin apioside, baicalin, oroxylin-A-7-O-fP-D-glucuronide and wogonoside) were identified from these active components by LC-Q/TOF-MS technology (liquid chromatography quadrupole time of flight tandem mass spectrometry). These 8 compounds may be the potential neuroprotective substances in Gegen Qinlian decoction.

An approach to screen nephrotoxic components in traditional Chinese medicine based on fluorescent probe and HK-2 cells and its application / 中国中药杂志

Using fluorescein diacetate labeled HK-2 cells, a fast method for screening nephrotoxic components in traditional Chinese medicines (TCMs) was proposed in this study. The methodology validation showed that the linearity, stability and accuracy of the proposed method were suitable for screening nephrotoxic components in vitro . This method was further applied on screen 352 components from 32 Chinese Pharmacopoeia-indexed toxic TCMs. The results indicated that 31 components from 14 toxic TCMs, including Badou, had significant toxicity on HK-2 cells, which suggested these components may cause nephrotoxicity. The components from the other 18 toxic TCMs had no significant toxicity on HK-2 cells.

Study on rapid screening and identifying hepatotoxic compounds in toosendan fructus / 中国中药杂志

Fluorescein diacetate-labeled HepG2 cells model and flouresence automatic microscopy screening assay were used for fast screening 23 components from Toosendan Fructus, in which 5 components showed significant toxicity on HepG2 cells. The 10 compounds in the 2 components were tentatively identified with LC-MS(n), and 3 of them (meliasenin B, trichilinin D and 1-O-tigloy-1-O-debenzoylohchinal) were prepared and identified. Further experiments showed that the 3 compounds displayed dose-dependent toxicity on HepG2 cells, suggesting that these compounds in Toosendan Fructus may cause hepatotoxicity.

Screening and identifying hepatotoxic components in Aucklandiae Radix with GC-MS / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To screen the potential hepatotoxic components in Chinese herb medicine Aucklandiae Radix.</p><p><b>METHODS</b>The potential hepatotoxic components were screened using HepG2 cells labeled with fluorescein diacetate from 25 fractions of Aucklandiae Radix, in which the hepatotoxic compounds were further identified with GC-MS.</p><p><b>RESULTS</b>Ten potential hepatotoxic fractions were screened. The identification results by GC-MS indicated that the main compounds in C09 were dehydrocostuslactone, santamarine (or magnolialide) and reynosin, and in C11 were α-costol and elemol.</p><p><b>CONCLUSION</b>Dehydrocostuslactone, santamarine (or magnolialide), reynosin, α-costol and elemol are potential hepatotoxic compounds in Aucklandiae Radix.</p>

Development of fluorescence imaging based assay for screening compounds with anti-migration activity / 药学学报

In the present study, A fluorescent imaging-based high-throughput screening method was developed for identifying anti-migratory compounds with 96-well Transwell plates. The correlation, precision and stability of this method were examined and the incubation time of dye Hoechst 33342 in addition to migration time was optimized. In addition, The inhibitory activity of anti-cancer drug paclitaxel on tumor cell migration was assayed and an IC50 value of 0.717 micromol x L(-1) was obtained. Using this method, 24 components from Rhizoma Alismatis were screened and one component with anti-migration activity was found. These results show that the new proposed method with good precision, stability and linear range has the potential to assay the inhibitory activity of anticancer compounds.

Development of fluorescence imaging-based system for screening compounds with antitumor activity / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To develop a fluorescence imaging-based novel system for quick screening of antitumor compounds in vitro.</p><p><b>METHODS</b>The antitumor activity of 26 components from Lindera aggregate were determined by relative number of viable cell labelled with fluorescein diacetate (FDA) in multiwell plates after exposure to these 26 different components. Then, the linearity and precision of this method were validated. The structures of active compounds in components with strong antitumor activity were deduced by LC/MS.</p><p><b>RESULTS</b>The linearity of this method for cells stained with FDA was validated (r² = 0.9858) in the range of 0-10⁴ cells per well, and the in-plate precision was 9.41 %. Two of 26 components from Lindera aggregate showed significant inhibition effect on proliferation of HepG2 cells (inhibition rate >90%).</p><p><b>CONCLUSION</b>This proposed rapid and reliable approach can be used for screening compounds with antitumor activity from Traditional Chinese Medicine in vitro. The major active compound of Lindera aggregate was putatively identified as norboldine by LC/MS analysis.</p>

Transport mechanism of danshensu across Caco-2 monolayer model / 中国中药杂志

<p><b>OBJECTIVE</b>To study the absorption mechanism of Danshensu by using Caco-2 monolayer model .</p><p><b>METHOD</b>Caco-2 cell monolayer model was used to study the bi-direction transport of Danshensu. An LC-MS method was developed to measure the concentration of Danshensu in cell culture medium and calculate the apparent permeability coefficients (P(app)). The effects of time, drug concentration and inhibitor on the absorption of Danshensu were studied.</p><p><b>RESULT</b>Transport of Danshensu was time and concentration dependent, and it was also effected by P-glycoprotein inhibitor. P(app) increased with time increase and tended to become saturated at some point. It, however,decreased while concentration of Danshensu increased. P(ratio) is larger than 1.5 . Verapamil can cause significantly effect on transport of Danshensu: P(app,A-B) increased and P(app,B-A) decreased .</p><p><b>CONCLUSION</b>The absorption of Danshensu in Caco-2 cell model may be an active transportation mediated by P-glycoprotein transporter.</p>

A method for predicting activity of traditional Chinese medicine based on quantitative composition-activity relationship of neural network model / 中国中药杂志

<p><b>OBJECTIVE</b>To study a method for evaluating the quality of traditional Chinese medicine (TCM) according as their activity.</p><p><b>METHOD</b>Combined with partial least squares (PLS), BP and RBF neural networks were selected to establish the model of quantitative composition-activity relationship (QCAR) due to their strong approximation capabilities for nonlinear function respectively. The activity of TCM was predicted with the QCAR model, and the quality of TCM was evaluated according to the predicted activity.</p><p><b>RESULT & CONCLUSION</b>The proposed method was applied to evaluate the quality of Chuanxiong. The results indicated that, in the indexes including training error, prediction error and correlation coefficient, the established model is better than the model established by principal component regression or PIS regression. The new model can accurately represent the complicated nonlinear relationship between the components and the bioactivity of Chuanxiong. Consequently, this method has potential to evaluate the quality of TCM according to bioactivity.</p>