| Based on syndrome differentiation and treatment,traditional Chinese medicine（TCM）treats diseases through a rational combination of various Chinese medicines.Compared with the single-component and single-target treatment of western medicine,TCM can realize multi-target comprehensive treatment of diseases through multiple components,making it show advantages in the treatment of complex diseases.Componentbased Chinese medicine（CCM）is a kind of modern Chinese medicine.Under the guidance of TCM and prescription-compatibility theory,CCM is generated by rational configuration of active ingredients.In the process of the development of CCM,the acquisition of the effective components of medicine materials usually depends on traditional separation methods.However,problems such as long-separation time,cumbersome steps,large consumption of organic reagents,the loss of active components have largely restricted the development of CCM.Therefore,developing a new separation technology of TCM with high efficiency,economical and environment-protecting performance can not only reduce the cost,but also realize the green and sustainable development of the CCM.Moreover,the intake of active ingredients in TCM mainly depends on gastrointestinal absorption,it is difficult to achieve targeted drug therapy.Loading the active ingredients of TCM on functional nanoparticles is conducive to the realization of targeted therapy and will further extend its application range.Anthraquinones are important active ingredients in TCM such as Rheum officinale and Polygonum cuspidatum,with purgative,anti-inflammatory,antibacterial,anti-tumor,blood pressure lowering and lipid lowering functions.The preparation of TCM containing anthraquinones by using functional magnetic nanoparticles has great application prospects in improving production efficiency of CCM.In this article,to realise effective separation of the anthraquinone components,Fe3O4magnetic nanoparticles synthesized by hydrothermal method were used as carrier of drug rapid separation,and using the character of the interaction force ofπ-πstacking between aromatic compounds,the planar alizarine（AZ）is designed to perform functional modification on three magnetic nanoparticles.The performance of three AZ-modified magnetic nanoparticles（Fe3O4-AZ,Fe3O4@Si O2-AZ,Fe3O4@Si O2-PEI-AZ）was evaluated,which can be used to realize the rapid separation of specific aromatic compounds in CCM.The adsorption performance of three nanoparticles to molecules with different aromatic compounds was investigated.Molecular-dynamics were used to simulate the changing trend of the adsorption-strength of aromatic compounds in different solvent conditions,and the rationality of the simulated data was verified by experiments.In order to investigate whether Fe3O4@Si O2-PEI-AZ can effectively adsorb anthraquinone compounds from complex ingredients,the Rheum officinale extract and actinomycetes fermentation products that produce anthraquinone-containing enediyne molecules were selected as the test objects,then the performance of Fe3O4@Si O2-PEI-AZ was evaluated.In order to achieve the purpose of targeted tumor therapy of anthraquinone CCM,functional nanomaterial Fe3O4@Si O2-PEI-FA-AZ with FR-targeting properties was designed.By targeting drug delivery to improve the local drug concentration of tumor cells,to enhance the anti-tumor effect.The research contents and results are as follows:1.Preparation and performance of three AZ-modified magnetic nanoparticles.Fe3O4 magnetic nanoparticles were modified which were synthesized by solvothermal synthesis,three kinds of Fe3O4 magnetic nanoparticles（Fe3O4-AZ,Fe3O4@Si O2-AZ,Fe3O4@Si O2-PEI-AZ）with different functional modifications were obtained.The surface shape,crystal form,bonding and magnetic of three kinds of Fe3O4@Si O2 magnetic nanoparticles were characterized by transmission electron microscope（TEM）,X-ray diffraction technique（XRD）,Fourier transform infrared spectroscopy（FTIR）and analysis in strengths of vibratory magnetic fields.It’s confirmed that the surface of Fe3O4 was successfully coated with Si O2,grafted on PEI and modified by AZ.The dispersibilities of three magnetic nanoparticles in 24 hours were compared,adsorption properties for three aromatic compounds of benzene ring were analyzed and evaluated.The results indicated that Fe3O4@Si O2-PEI-AZ shows the best dispersion and highest load capacity as most particles remained uniformly dispersed after standing in aqueous solution for 24 hours;in 5%methanol-water solution（v/v）,the loading capacity of three aromatic compounds of benzene ring class is up to 102 mg/g.Fe3O4@Si O2-PEI-AZ was selected as the carrier in follow research because of the best dispersion and highest load capacity.2.The study of Fe3O4@Si O2-PEI-AZ on adsorption of monomer active components of TCMThe adsorption properties of benzene ring aromatic compounds containing one,two and three benzene rings were explored.First,Three-Dimensional model（TD-model）of Fe3O4@Si O2-PEI-AZ was constructed through computer-aided design（CAD）which was used to simulate different solvent conditions to get adsorption effect of Fe3O4@Si O2-PEI-AZ on various components.The adsorption of Fe3O4@Si O2-PEI-AZ with aromatic compound molecules is mainly realized throughπ-πinteraction in aqueous solution;Theπ-πinteraction of Fe3O4@Si O2-PEI-AZ with aromatic components is destroyed to desorb in methanol solution;compounds with three aromatic benzene rings were adsorbed closely while components with one or two aromatic benzene rings were weakly binding by Fe3O4@Si O2-PEI-AZ in 20%methanol-water solution（v/v）.Then the adsorptions in various methanol-water solution were compared through the designed experiment.The results indicated that adsorption trends of components with the same structure type by Fe3O4@Si O2-PEI-AZ were similar.Compounds with two or three aromatic benzene rings were adsorbed while components one aromatic benzene ring were almost not adsorbed by Fe3O4@Si O2-PEI-AZ in 5%methanol-water solution（v/v）.The adsorption specificity of aromatic compounds by F Fe3O4@Si O2-PEI-AZ increased gradually with the methanol concentration increasing.Only the aromatic compounds with three benzene rings,alizarin,emodin and rhein,can be adsorbed by Fe3O4@Si O2-PEI-AZ in 55%methanol-water solution（v/v）,while compounds with two benzene rings,naphthalene,1-naphthol and kaempferol were not adsorbed,which indicated that the components with anthraquinone structure can be specifically separated under the condition of high concentration of organic solvents.At last,the rate of reused Fe3O4@Si O2-PEI-AZ was explored.The adsorption rates of emodin were above 95%in 10 consecutive uses,which indicated the high recycling rate and the ability of recycling of Fe3O4@Si O2-PEI-AZ.From the experiments which the influence on the stability of adsorbed compounds by Fe3O4@Si O2-PEI-AZ,it indicated that the structures and properties of the adsorbed compounds were not affected by the adsorption process.3.Adsorption and separation of anthraquinone compounds from TCM and fermented extractsBased on adsorption differences of aromatic compounds by functional magnetic nanomaterials in different methanol concentrations.Fe3O4@Si O2-PEI-AZ was used for the effective separation of anthraquinone components in rhubarb extract and anthraquinone-fused enediynes（TNMs）in actinomycete fermentation products.In 25%methanol-water solution（v/v）,four anthraquinone components were isolated and enriched from the extract of rhubarb by magnetic specific adsorption method of Fe3O4@Si O2-PEI-AZ,such as rhein,emodin,chrysophanol,physcion,etc.In 10%methanol-water solution（v/v）,Fe3O4@Si O2-PEI-AZ was used to purify TNMs from actinomycete fermentation products.HPLC was used to demonstrate that specific adsorption of TNMs components by Fe3O4@Si O2-PEI-AZ.The interference of fatty acids and other components in fermentation products was solved and the separation steps of TNMs were simplified.It is proved that the magnetic nanomaterial Fe3O4@Si O2-PEI-AZ modified by Alizarin can be used for the rapid and specific separation of anthraquinone structures in the mixed system through the adsorption and separation experiments of rhubarb extract and anthraquinone enediyne fermentation extract.Compared with the traditional separation method,Magnetic specific separation method of Alizarin modified magnetic nanoparticles,Fe3O4@Si O2-PEI-AZ,which can selectively adsorb anthraquinones in complex systems to achieve enrichment and separation.With the advantages of simple,quick and less consumption of organic reagents,the method enriches the research on separation of pharmacodynamic components.4.The study on folic acid（FA）targeting functional magnetic nanomaterials loaded with emodin for tumor targeting therapyFolate receptor（FR）is widely expressed on the surface of various tumor cells,but only a little on the surface of normal cells,so FA was designed as the anchoring agent for tumor cells,magnetic nanoparticles modified with AZ were used as adsorbents for anthraquinones,magnetic nanoparticles（Fe3O4@Si O2-PEI-FA-AZ）co-modified by FA and AZ were used for targeted delivery of anthraquinone drugs.Research showed that emodin can enhance the inhibitory effect on tumor cell proliferation in vitro and enhance the effect of inducing tumor cell apoptosis.However,due to poor water solubility and low bioavailability of emodin,the drug concentration in the body could not reach the effective anti-tumor level during drug administration,which limited the further application of emodin.Therefore,Fe3O4@Si O2-PEI-FA-AZ was used to load emodin to achieve tumor targeted therapy of emodin.FR expression levels in Human cervical cancer cells（Hela）,Rat myocardial cells（H9C2）,human oral epidermoid cancer cells（KB）,mammary carcinoma cells（4T1）,human breast cancer cells（MCF7）,mouse fibroblasts（NIH-3T3）,human ovarian carcinoma cells（SKOV3）were compared.The results of Western Blot showed that compared with H9C2,4T1 and NIH-3T3 cells,the expression of FR on the cell surface of Hela,KB,MCF7 and SKOV3 cells was significantly increased,it was confirmed that abnormal expression of FR in a variety of tumor cells.Low expression FR 4T1 cells were used as control to verify the anti-tumor effect of Fe3O4@Si O2-PEI-FA-AZ loaded drugs.Thiazolyl blue tetrazolium（MTT）test was performed on tumor cells KB and MCF7 with high expression of FR.The anti-tumor effects of free emodin,emodin loaded with untargeted material（Fe3O4@Si O2-PEI-AZ）and FA targeted material（Fe3O4@Si O2-PEI-FA-AZ）were determined.The results indicated that compared with free emodin and emodin loaded Fe3O4@Si O2-PEI-AZ,emodin loaded Fe3O4@Si O2-PEI-FA-AZ had stronger antitumor effect on KB and MCF7.The anti-tumor effect of targeted emodin treatment group on Hela and KB was significantly enhanced than 4T1 of FR low expression. |
PDF Full Text Request