Construction and application of covalently bonded CD147 cell membrane chromatography model based on polystyrene microspheres.

In this study, a novel cell membrane chromatography (CMC) model was developed to investigate cluster of differentiation 147 (CD147) targeted anti-tumor drug leads for specific screening and ligand-receptor interaction analysis by SNAP-tagged CD147 fusion protein conjugation and polystyrene microspheres (PS) modification. Traditional Chinese medicines (TCMs) are widely used in the treatment of cancer. CD147 plays important roles in tumor progression and acts as an attractive target for therapeutic intervention; therapeutic drugs for CD147-related cancers are limited to date. Thus, a screening method for active components in TCMs is crucial for the further research and development of CD147 antagonists. However, improvement is still needed to perform specific and accurate drug lead screening using the CMC-based method. Recently, our group developed a covalently immobilized receptor-SNAP-tag/CMC model using silica gel as carrier. Besides the carboxyl group on multi-step modified silica particles, the amino group of benzyl-guanine (BG, substrate of SNAP-tag) also possesses reactivity towards the carboxyl group on available carboxyl-modified PS. Herein, we used PS as carrier and an extended SNAP-tag with CD147 receptor to construct the PS-BG-CD147/CMC model for active compound investigation coupled with HPLC/MS and applied this coupled PS-BG-CD147/CMC-HPLC/MS two-dimensional system to drug lead screening from Nelumbinis Plumula extract (NPE) sample. In addition, to comprehensively verify the pharmacological effects of screened ingredients, a cell proliferation inhibition assay was performed, and the interaction between the ingredients and CD147 was studied by the frontal analysis method. This study developed a high-throughput PS-based CMC screening platform, which could be widely applied and utilized in chromatographic separation and drug lead discovery.

Molecular Mechanism of Nelumbinis Plumula Alkaloids in Prevention and Treatment of Non-Small Cell Lung Cancer： an Analysis Based on Network Pharmacology and Cell Experiment / 中国实验方剂学杂志

Objective:To explore the potential molecular mechanism of Nelumbinis Plumula alkaloids （NAPs） in the prevention and treatment of non-small cell lung cancer （NSCLC） based on network pharmacology and cell experiment. Method:The main active components of NAPs were obtained by searching Traditional Chinese Medicine System Pharmacology Database and Analysis Platform （TCMSP） and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine （BATMAN-TCM）， and their main targets were predicted and analyzed by employing Swiss Target Prediction. The main target genes of NSCLC were retrieved from GeneCards， Online Mendelian Inheritance in Man （OMIM） and DrugBank databases. The resulting common targets were imported into STRING platform for constructing the protein-protein interaction （PPI） network， followed by gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis based on Database for Annotation， Visualization， and Integrated Discovery （DAVID）. The NAPs-common target -pathway network was constructed by Cytoscape 3.7.1. After NSCLC cell line A549 was treated with isoliensinine， the cell morphology was observed under an inverted fluorescence microscope. The effect of isoliensinine on A549 vitality was detected by cell counting kit-8 （CCK-8） assay and the target protein changes were verified by Western blot. Result:The main active components for NAPs against NSCLC were lysicamine， liensinine， and isoliensinine. The phosphatidylinositol-3-kinase-protein kinase B （PI3K-AKT）， RAS-related protein 1 （Rap1）， epidermal growth factor family of receptor tyrosine kinases （ErbBs）， and hypoxia inducible factor-1 （HIF-1） pathways were mainly involved for binding adenosine triphosphate （ATP） and regulating protein kinase activity. The main targets included protein kinase B-1 （AKT1）， alpha catalytic subunit of phosphoinositol-3-kinase （PIK3CA）， cyclin-dependent kinase 2 （CDK2）， mitogen-activated protein kinase-1 （MAPK1）， epidermal growth factor receptor （EGFR）， adenosine triphosphate-binding cassette B1 （ABCB1）， mammalian target of rapamycin （mTOR）， tyrosine kinase （Src）， Janus kinase 1 （JAK1）， and G1-phase-specific gene cyclin-D₁ （CCND1）. The <italic>in vitro</italic> cell experiment also revealed that isoliensinine down-regulated the expression of phosphorylated AKT （p-AKT） and phosphorylated mTOR （p-mTOR） in a concentration- and time-dependent manner and inhibited the growth of A549 cells. Conclusion:NAPs exert the preventive and therapeutic effects against NSCLC through multiple components， multiple targets， and multiple pathways， especially the PI3K-AKT pathway.

[Pharmacological mechanism of traditional sedative effect of alkaloids in Nelumbinis Plumula based on network pharmacology].

Nelumbinis Plumula has the traditional sedative effect,but its mechanism is unclear. In this study,the relationship between traditional sedative effect and hypnotic effect of Nelumbinis Plumula was taken as the starting point to study the hypnotic mechanism of the major medicinal components in Nelumbinis Plumula by the network pharmacology method. Targets of active Nelumbinis Plumula alkaloids were screened by Swiss Target Prediction server,TCMSP and BATMAN-TCM. Targets of hypnotic drugs approved by FDA were screened from Drug Bank,OMIM,TTD databases. The common targets were screened by GO and KEGG pathways. Cytoscape 3. 7. 1 software was used to construct the network of " active component-target-pathway-disease". The results of network analysis showed that 21 active compounds were associated with 44 targets and 28 pathways. Among them,21 compounds,35 targets and 15 pathways were predicted to be related to sedative hypnosis. Nelumbinis Plumula showed the hypnotic effect by acting on neuroactive ligand-receptor interaction pathway,regulation of actin cytoskeleton pathway,calcium signaling pathway,cholinergic synapse pathway.This study preliminarily revealed the potential active compounds and possible mechanisms of traditional sedative effect of Nelumbinis Plumula,which provided a theoretical basis for further experimental studies on medicinal materials and its mechanisms.

Evaluation of different types of bitterness masking on inhibition of bitterness efficiency and law of four Chinese materia medicine decoctions of Gentianae Radix et Rhizoma, Sophorae Flavescentis Radix, Andrographis Herba, and Nelumbinis Plumula / 中草药

Objective To investigate the effects and the law of anti bitterness efficiency of hydroxypropyl-β-cyclodextrin (HP-β-CD) and other different types bitterness masking at different concentrations on the bitterness decoction of Chinese materia medica (BDCMM) by using the traditional human taste panel method (THTPM) and Electronic tongue (E-tongue). Methods Based on THTPM, the bitterness reduction value (ΔI), the correction rate of bitterness suppression (CRBS), and the correction rate of bitterness suppression potency index (PI50) were used as indicators to evaluate the bitterness suppression effect of four bitterness masking at different concentrations on BDCMM such as Sophorae Flavescentis Radix (SFR), and the relationship model between the concentrations of ΔI and bitterness masking was established to explore the bitterness suppression rule of bitterness masking on different BDCMM. Based on the E-tongue method, the E-tongue taste response information values of HP-β-CD and Acesulfame K to BDCMM such as SFR were measured at different concentrations, and the electronic tongue bitterness reduction value (ΔIe) was used as an index to explore the change rule of ΔIe with bitterness masking concentration. THTPM and E-tongue was combined to establish a prediction model of the effect of bitterness suppression. Results The ΔI, CRBS, and PI50 of four kinds of BDCMM with different concentrations of bitterness masking were measured, which can be used to compare the bitterness suppression effect of bitterness masking; The relation between ΔI and bitterness masking concentration accorded with Weibull curve model; Based on E-tongue, ΔIe was obtained, and the model of the relationship between ΔIe and bitterness masking concentration was established. The relationship between ΔIe and ΔI was established. The model determination coefficients of HP-β-CD for the two types of ΔIe of the two BDCMM were 0.986 1, 0.977 9, 0.989 0, and 0.982 0 respectively (P < 0.01, n = 6). Acesulfame had no response to the sensor and did not establish a model of bitterness suppression law. Conclusion Based on THTPM and E-tongue, a method for evaluating the bitterness suppression effect of bitterness masking on BDCMM was established. The bitterness suppression effect of bitterness masking on BDCMM such as SFR was studied by molecular inclusion, high-efficiency sweetening, and other bitterness suppression mechanisms were inverstigated.

[Studies on the chemical components of Nelumbinis Plumula and the inhibitory activity on protein disulfide isomerase].

Increasing evidence suggested that protein disulfide isomerase supported the survival and progression of several cancers. Nelumbinis Plumula is a Chinese traditional herb which showed antitumor activity. To find if the Nelumbinis Plumula affect protein disulfide isomerase activity, we studied its chemical constituents, and 12 monomeric compounds were isolated by means of solvent extraction, silica gel column chromatography, preparative HPLC and recrystallization. Among them, N-methylcoclaurine, kaempferol, chrysoeriol-7-O-neohesperidoside and mannitol were obtained for the first time. Following, we tested the compounds inhibitory activity on protein disulfide isomerase. The results showed that N-methylcoclaurine, neferine, liensinine and isoliensinine could inhibit the activity of protein disulfide isomerase in vitro, their IC50 values were 1.4, 2.9, 4.0 and 5.4 µmol•L⁻¹, respectively.