CN113577040A - Nano-carrier loaded with traditional Chinese medicine monomers and application of nano-carrier in preparation of antitumor drugs - Google Patents
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Abstract
The invention discloses a nano-carrier loaded with traditional Chinese medicine monomers and application thereof in preparing antitumor drugs, comprising the following steps: preparing a nano carrier material; purifying the carrier material and preparing nanoparticles; extracting anti-tumor Chinese medicine monomers, and carrying out different fluorescence labeling on different Chinese medicine molecules; introducing monomer molecules of Chinese medicinal materials into nanoparticles; detecting the introduction condition of the traditional Chinese medicine monomer molecules, and screening the nanoparticles; preparing a carrier solution and a nano preparation; introducing the nano preparation into living tumor cells; tumor cell morphology and number were observed and the results were recorded. The invention can research the effect of the nanometer preparation added with the anti-tumor traditional Chinese medicine monomer molecules, enables the nanometer preparation added with different traditional Chinese medicine monomer molecules to act on living tumor cells, and can know the effect of the nanometer carrier co-loading the anti-tumor medicine and the nucleic acid medicine on the tumor cells by observing the proliferation condition of the tumor cells and the shape of the tumor cells.
Description
Technical Field
The invention belongs to the technical field of biomedicine, and particularly relates to a nano carrier loaded with a traditional Chinese medicine monomer and application of the nano carrier in preparation of an anti-tumor medicine.
Background
Nanoscale Drug Carriers (Nanoscale Drug Carriers) are a submicron Drug carrier delivery system that belongs to the Nanoscale micro-category. The drug is encapsulated in submicron particles, so that the release speed can be adjusted, the permeability of a biological membrane is increased, the distribution in a body is changed, the bioavailability is improved, and the like. Nanoparticles (NPs), also known as nanoparticles, are solid colloidal particles of 10-1000 nm in size, generally composed of natural or synthetic polymeric substances, and can be used as carriers for conducting or delivering drugs. Due to differences in materials and manufacturing processes, nanospheres (nanospheres) and nanocapsules (nanocapsules), both collectively referred to as nanoparticles, can be formed. Solid lipid nanoparticles (solid lipid nanoparticles SLN) are a novel nanoparticle drug delivery system which is developed in recent years, wherein solid lipid (natural or synthetic) is used as a carrier, and a drug is wrapped in a lipid core to form solid colloidal particles, and the particle diameter is between 50 and 1000 nm.
The existing antitumor drug nano preparation only carries a single antitumor drug and cannot carry traditional Chinese medicine monomers or traditional Chinese medicine molecules, so that the killing effect on tumors is limited and the treatment effect is influenced in the actual treatment process, and the screening of antitumor traditional Chinese medicines is also the key point of research in the field.
Disclosure of Invention
The invention aims to overcome the defects in the prior art, and provides a nano carrier loaded with traditional Chinese medicine monomers and application of the nano carrier in preparation of antitumor drugs, so as to solve the problem that the effect of the antitumor drug nano preparation provided in the background technology is poor in the treatment process.
In order to achieve the purpose, the invention provides the following technical scheme: the nano carrier loaded with the traditional Chinese medicine monomer and the application thereof in preparing the anti-tumor medicine comprise the following components: 16-18 parts of black nightshade, 14-16 parts of fructus polygoni orientalis, 12-14 parts of phellinus linteus, 8-10 parts of caragana sinica linn root, 13-15 parts of curcuma zedoary, 6-8 parts of paris polyphylla, 5-7 parts of solanum lyratum, 8-10 parts of gynostemma pentaphylla, 8-10 parts of sculellaria barbata and 8-10 parts of aloe.
The application method of the nano-carrier loaded with the traditional Chinese medicine monomers and the application of the nano-carrier in preparing the anti-tumor medicine comprises the following steps:
the method comprises the following steps: preparing a nano carrier material;
step two: purifying the carrier material and preparing nanoparticles;
step three: extracting anti-tumor Chinese medicine monomers, and carrying out different fluorescence labeling on different Chinese medicine molecules;
step four: introducing monomer molecules of Chinese medicinal materials into nanoparticles;
step five: detecting the introduction condition of the traditional Chinese medicine monomer molecules, and screening the nanoparticles;
step six: preparing a carrier solution and a nano preparation;
step seven: introducing the nano preparation into living tumor cells;
step eight: tumor cell morphology and number were observed and the results were recorded.
Compared with the prior art, the invention provides the nano-carrier loaded with the traditional Chinese medicine monomers and the application thereof in preparing the anti-tumor drugs, and the nano-carrier has the following beneficial effects:
1. according to the invention, through setting different control group experiments, the effect of the nano preparation added with the anti-tumor Chinese medicine monomer molecules can be respectively researched, the nano preparation added with different Chinese medicine monomer molecules acts on living tumor cells, and the effect of the nano carrier loaded with the anti-tumor medicine and the nucleic acid medicine on the tumor cells can be known through observing the proliferation condition and the morphology of the tumor cells;
2. according to the invention, the nano-carrier added with the traditional Chinese medicine monomers is detected by adopting a fluorescence labeling technical means, so that a worker can screen the nano-carrier, the nano-carrier added with the medicine successfully can be selected, and the accuracy of the result is increased.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides the following technical scheme: the nano carrier loaded with the traditional Chinese medicine monomer and the application thereof in preparing the anti-tumor medicine comprise the following components: 16 parts of black nightshade, 14 parts of fructus polygoni orientalis, 12 parts of phellinus linteus, 8 parts of caragana sinica root, 13 parts of curcuma zedoary, 6 parts of paris polyphylla, 5 parts of solanum lyratum, 8 parts of gynostemma pentaphylla, 8 parts of sculellaria barbata and 8 parts of aloe.
The application method of the nano-carrier loaded with the traditional Chinese medicine monomers and the application of the nano-carrier in preparing the anti-tumor medicine comprises the following steps:
the method comprises the following steps: preparing a nano carrier material;
step two: purifying the carrier material and preparing nanoparticles;
step three: extracting anti-tumor Chinese medicine monomers, and carrying out different fluorescence labeling on different Chinese medicine molecules;
step four: introducing monomer molecules of Chinese medicinal materials into nanoparticles;
step five: detecting the introduction condition of the traditional Chinese medicine monomer molecules, and screening the nanoparticles;
step six: preparing a carrier solution and a nano preparation;
step seven: introducing the nano preparation into living tumor cells;
step eight: tumor cell morphology and number were observed and the results were recorded.
In the present invention, preferably, in the step one, a proper amount of cysteine molecular compound is selected, a reducing agent is added into the molecular compound to reduce and crack cysteine to lipoic acid, and the reacted lipoic acid is extracted.
In the invention, preferably, in the second step, a proper amount of lipoic acid is taken and dissolved in methanol, cysteine hydrochloride is added for reaction for 6 hours, redundant methanol is removed, diluted hydrochloric acid and water are respectively used for washing for a plurality of times, impurities are further removed, the solution after the impurities are removed is dissolved in chloroform, 1% sodium cholate solution is added for ultrasonic emulsification, and nanoparticles are obtained after the reaction.
In the invention, preferably, in the third step, a proper amount of antitumor drug such as chlorambucil is selected, a proper amount of Chinese medicine monomer molecules are respectively extracted, and the Chinese medicine monomer molecules are subjected to element labeling by adopting a fluorescence labeling technical means.
In the invention, preferably, in the fourth step, the marked traditional Chinese medicine monomer molecules are introduced into the nanoparticles, and different control group experiments are set;
in the invention, preferably, in the fifth step, the nanoparticles in the experimental group are subjected to fluorescence detection, and the nanoparticles in which the traditional Chinese medicine monomer molecules are successfully introduced are screened out according to the detection result.
In the present invention, preferably, in the sixth step, large particulate matters are removed by using a centrifugal technique, and then sodium cholate and other matters are removed by ultrafiltration, so as to obtain pure nano materials introduced with monomer molecules of traditional Chinese medicines.
In the present invention, preferably, in step seven, a suitable living tumor cell is selected, the tumor cell is placed in a corresponding culture dish, the nanomaterial introduced with the traditional Chinese medicine monomer molecule is introduced into the tumor cell, and the proliferation of the tumor cell is waited.
In the present invention, it is preferable that, in step eight, after cell proliferation, the number of tumor cell proliferations and the morphology of tumor cells are observed under a microscope, data results are recorded, and data analysis is performed on experimental results.
Example one
The nano carrier loaded with the traditional Chinese medicine monomer and the application thereof in preparing the anti-tumor medicine comprise the following components: 16 parts of black nightshade, 14 parts of fructus polygoni orientalis, 12 parts of phellinus linteus, 8 parts of caragana sinica root, 13 parts of curcuma zedoary, 6 parts of paris polyphylla, 5 parts of solanum lyratum, 8 parts of gynostemma pentaphylla, 8 parts of sculellaria barbata and 8 parts of aloe.
The application method of the nano-carrier loaded with the traditional Chinese medicine monomers and the application of the nano-carrier in preparing the anti-tumor medicine comprises the following steps:
the method comprises the following steps: preparing a nano carrier material;
step two: purifying the carrier material and preparing nanoparticles;
step three: extracting anti-tumor Chinese medicine monomers, and carrying out different fluorescence labeling on different Chinese medicine molecules;
step four: introducing monomer molecules of Chinese medicinal materials into nanoparticles;
step five: detecting the introduction condition of the traditional Chinese medicine monomer molecules, and screening the nanoparticles;
step six: preparing a carrier solution and a nano preparation;
step seven: introducing the nano preparation into living tumor cells;
step eight: tumor cell morphology and number were observed and the results were recorded.
In the present invention, preferably, in the step one, a proper amount of cysteine molecular compound is selected, a reducing agent is added into the molecular compound to reduce and crack cysteine to lipoic acid, and the reacted lipoic acid is extracted.
In the invention, preferably, in the second step, a proper amount of lipoic acid is taken and dissolved in methanol, cysteine hydrochloride is added for reaction for 6 hours, redundant methanol is removed, diluted hydrochloric acid and water are respectively used for washing for a plurality of times, impurities are further removed, the solution after the impurities are removed is dissolved in chloroform, 1% sodium cholate solution is added for ultrasonic emulsification, and nanoparticles are obtained after the reaction.
In the invention, preferably, in the third step, a proper amount of antitumor drug such as chlorambucil is selected, a proper amount of Chinese medicine monomer molecules are respectively extracted, and the Chinese medicine monomer molecules are subjected to element labeling by adopting a fluorescence labeling technical means.
In the present invention, preferably, in the fourth step, the labeled monomer molecules of the traditional Chinese medicine are introduced into the nanoparticles, and different control group experiments are set.
In the invention, preferably, in the fifth step, the nanoparticles in the four experimental groups are subjected to fluorescence detection, and the nanoparticles in which the traditional Chinese medicine monomer molecules are successfully introduced are screened out according to the detection result.
In the present invention, preferably, in the sixth step, large particulate matters are removed by using a centrifugal technique, and then sodium cholate and other matters are removed by ultrafiltration, so as to obtain pure nano materials introduced with monomer molecules of traditional Chinese medicines.
In the present invention, preferably, in step seven, a suitable living tumor cell is selected, the tumor cell is placed in a corresponding culture dish, the nanomaterial introduced with the traditional Chinese medicine monomer molecule is introduced into the tumor cell, and the proliferation of the tumor cell is waited.
In the present invention, it is preferable that, in step eight, after cell proliferation, the number of tumor cell proliferations and the morphology of tumor cells are observed under a microscope, data results are recorded, and data analysis is performed on experimental results.
Example two
The nano carrier loaded with the traditional Chinese medicine monomer and the application thereof in preparing the anti-tumor medicine comprise the following components: 17 parts of black nightshade, 15 parts of fructus polygoni orientalis, 13 parts of phellinus linteus, 9 parts of caragana sinica root, 14 parts of curcuma zedoary, 7 parts of paris polyphylla, 6 parts of solanum lyratum, 9 parts of gynostemma pentaphylla, 9 parts of sculellaria barbata and 9 parts of aloe;
the application method of the nano-carrier loaded with the traditional Chinese medicine monomers and the application of the nano-carrier in preparing the anti-tumor medicine comprises the following steps:
the method comprises the following steps: preparing a nano carrier material;
step two: purifying the carrier material and preparing nanoparticles;
step three: extracting anti-tumor Chinese medicine monomers, and carrying out different fluorescence labeling on different Chinese medicine molecules;
step four: introducing monomer molecules of Chinese medicinal materials into nanoparticles;
step five: detecting the introduction condition of the traditional Chinese medicine monomer molecules, and screening the nanoparticles;
step six: preparing a carrier solution and a nano preparation;
step seven: introducing the nano preparation into living tumor cells;
step eight: tumor cell morphology and number were observed and the results were recorded.
In the present invention, preferably, in the step one, a proper amount of cysteine molecular compound is selected, a reducing agent is added into the molecular compound to reduce and crack cysteine to lipoic acid, and the reacted lipoic acid is extracted.
In the invention, preferably, in the second step, a proper amount of lipoic acid is taken and dissolved in methanol, cysteine hydrochloride is added for reaction for 6 hours, redundant methanol is removed, diluted hydrochloric acid and water are respectively used for washing for a plurality of times, impurities are further removed, the solution after the impurities are removed is dissolved in chloroform, 1% sodium cholate solution is added for ultrasonic emulsification, and nanoparticles are obtained after the reaction.
In the invention, preferably, in the third step, a proper amount of antitumor drug such as chlorambucil is selected, a proper amount of Chinese medicine monomer molecules are respectively extracted, and the Chinese medicine monomer molecules are subjected to element labeling by adopting a fluorescence labeling technical means.
In the present invention, preferably, in the fourth step, the labeled monomer molecules of the traditional Chinese medicine are introduced into the nanoparticles, and different control group experiments are set.
In the invention, preferably, in the fifth step, the nanoparticles in the four experimental groups are subjected to fluorescence detection, and the nanoparticles in which the traditional Chinese medicine monomer molecules are successfully introduced are screened out according to the detection result.
In the present invention, preferably, in the sixth step, large particulate matters are removed by using a centrifugal technique, and then sodium cholate and other matters are removed by ultrafiltration, so as to obtain pure nano materials introduced with monomer molecules of traditional Chinese medicines.
In the present invention, preferably, in step seven, a suitable living tumor cell is selected, the tumor cell is placed in a corresponding culture dish, the nanomaterial introduced with the traditional Chinese medicine monomer molecule is introduced into the tumor cell, and the proliferation of the tumor cell is waited.
In the present invention, it is preferable that, in step eight, after cell proliferation, the number of tumor cell proliferations and the morphology of tumor cells are observed under a microscope, data results are recorded, and data analysis is performed on experimental results.
EXAMPLE III
The nano carrier loaded with the traditional Chinese medicine monomer and the application thereof in preparing the anti-tumor medicine comprise the following components: 18 parts of black nightshade, 16 parts of fructus polygoni orientalis, 14 parts of phellinus linteus, 10 parts of caragana sinica root, 15 parts of curcuma zedoary, 8 parts of paris polyphylla, 7 parts of solanum lyratum, 10 parts of gynostemma pentaphylla, 10 parts of sculellaria barbata and 10 parts of aloe.
The application method of the nano-carrier loaded with the traditional Chinese medicine monomers and the application of the nano-carrier in preparing the anti-tumor medicine comprises the following steps:
the method comprises the following steps: preparing a nano carrier material;
step two: purifying the carrier material and preparing nanoparticles;
step three: extracting anti-tumor Chinese medicine monomers, and carrying out different fluorescence labeling on different Chinese medicine molecules;
step four: introducing monomer molecules of Chinese medicinal materials into nanoparticles;
step five: detecting the introduction condition of the traditional Chinese medicine monomer molecules, and screening the nanoparticles;
step six: preparing a carrier solution and a nano preparation;
step seven: introducing the nano preparation into living tumor cells;
step eight: tumor cell morphology and number were observed and the results were recorded.
In the present invention, preferably, in the step one, a proper amount of cysteine molecular compound is selected, a reducing agent is added into the molecular compound to reduce and crack cysteine to lipoic acid, and the reacted lipoic acid is extracted.
In the invention, preferably, in the second step, a proper amount of lipoic acid is taken and dissolved in methanol, cysteine hydrochloride is added for reaction for 6 hours, redundant methanol is removed, diluted hydrochloric acid and water are respectively used for washing for a plurality of times, impurities are further removed, the solution after the impurities are removed is dissolved in chloroform, 1% sodium cholate solution is added for ultrasonic emulsification, and nanoparticles are obtained after the reaction.
In the invention, preferably, in the third step, a proper amount of antitumor drug such as chlorambucil is selected, a proper amount of Chinese medicine monomer molecules are respectively extracted, and the Chinese medicine monomer molecules are subjected to element labeling by adopting a fluorescence labeling technical means.
In the present invention, preferably, in the fourth step, the labeled monomer molecules of the traditional Chinese medicine are introduced into the nanoparticles, and different control group experiments are set.
In the invention, preferably, in the fifth step, the nanoparticles in the four experimental groups are subjected to fluorescence detection, and the nanoparticles in which the traditional Chinese medicine monomer molecules are successfully introduced are screened out according to the detection result.
In the present invention, preferably, in the sixth step, large particulate matters are removed by using a centrifugal technique, and then sodium cholate and other matters are removed by ultrafiltration, so as to obtain pure nano materials introduced with monomer molecules of traditional Chinese medicines.
In the present invention, preferably, in step seven, a suitable living tumor cell is selected, the tumor cell is placed in a corresponding culture dish, the nanomaterial introduced with the traditional Chinese medicine monomer molecule is introduced into the tumor cell, and the proliferation of the tumor cell is waited.
In the present invention, it is preferable that, in step eight, after cell proliferation, the number of tumor cell proliferations and the morphology of tumor cells are observed under a microscope, data results are recorded, and data analysis is performed on experimental results.
In conclusion, through the experimental results, it is shown that,
the nanometer preparation carrying both antitumor medicine and nucleic acid medicine has great killing capacity on tumor cell, and the nanometer preparation carrying both antitumor medicine and nucleic acid medicine can damage tumor cell.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (2)
1. The nano carrier loaded with the traditional Chinese medicine monomer and the application thereof in preparing anti-tumor medicines are characterized in that: comprises the following components: 16-18 parts of black nightshade, 14-16 parts of fructus polygoni orientalis, 12-14 parts of phellinus linteus, 8-10 parts of caragana sinica linn root, 13-15 parts of curcuma zedoary, 6-8 parts of paris polyphylla, 5-7 parts of solanum lyratum, 8-10 parts of gynostemma pentaphylla, 8-10 parts of sculellaria barbata and 8-10 parts of aloe.
2. The nano-carrier loaded with traditional Chinese medicine monomers and the application method thereof in preparing anti-tumor drugs according to claim 1 are characterized in that: the method comprises the following steps:
the method comprises the following steps: preparing a nano carrier material;
step two: purifying the carrier material and preparing nanoparticles;
step three: extracting anti-tumor Chinese medicine monomers, and carrying out different fluorescence labeling on different Chinese medicine molecules;
step four: introducing monomer molecules of Chinese medicinal materials into nanoparticles;
step five: detecting the introduction condition of the traditional Chinese medicine monomer molecules, and screening the nanoparticles;
step six: preparing a carrier solution and a nano preparation;
step seven: introducing the nano preparation into living tumor cells;
step eight: tumor cell morphology and number were observed and the results were recorded.
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