CN103070983B - Composition based on main components of Pu'er tea and application thereof - Google Patents
Composition based on main components of Pu'er tea and application thereof Download PDFInfo
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Abstract
The invention discloses composition based on main components of Pu'er tea and the application thereof. The composition comprises tea brown pigment, tea polysaccharide, tea polyphenol, caffeine, gamma-aminobutyric acid and gallic acid. A wild type mouse embryo fibroblast is utilized to test the killing effect of DNA toxicity chemical substance to normal cells and the effect that the composition provided by the invention protects the normal cells from DNA damage and prevents apoptosis; the result shows that the composition provided by the invention can effectively prevent the normal cell death caused by DNA toxicity chemical substance; and further research shows that the composition provided by the invention does not protect tumor cell death caused by the DNA toxicity chemical compound, conversely, the composition provided by the invention can promote the killing effect of the DNA toxicity chemical compound to the tumor cells. Therefore, the composition can be used for protecting the normal cells during the chemotherapy of tumor patients and alleviating toxic side effects caused by tumor chemotherapy, and meanwhile sensibilizes chemotherapy drug, enhances chemotherapy effect, and improves survival quality of chemotherapy patients.
Description
Technical field
The present invention relates to a kind of compositions based on Folium camelliae assamicae main component and application thereof, more particularly, the present invention relates to the application of said composition in the chemical substance biohazard of prevention DNA toxicity.
Background technology
The chemical substance of DNA toxicity or genotoxicity (genotoxic), refers to that a large class can cause DNA damage, and then causes gene mutation, cell death or tumorigenic chemical substance.Wherein comprise and be directly attached on DNA, chemical modification is carried out to DNA, causes DNA damage; Or act on DNA in conjunction with associated protein, damage etc. is caused to DNA.Major part teratogenesis, cause prominent, carcinogenic compound and all belong to this class, this compounds is also often run in people's daily life, as vehicle exhaust, and smoke from cigarette, kitchen fume etc.
Due to the multiplication capacity of tumor cell Showed Very Brisk, in cell, DNA replication dna is very active, also becomes the major target class of this kind of determining presence of genotoxic compound.At present, the tumor chemotherapeutic drug generally applied clinically, as cisplatin and derivant carboplatin thereof, cyclophosphamide, amycin etc. is all the compound of this class.Because these compounds also have damage to Normocellular DNA, the tumour patient accepting chemotherapy there will be serious toxic and side effects usually, as leukopenia, vomiting, diarrhoea etc.
Therefore, the bio-toxicity effect of DNA toxic chemical to the mankind more and more gets more and more people's extensive concerning, and how to prevent its teratogenesis, causes prominent, that carcinogenesis has also become people's extensive concern topic.Clinically, the toxic and side effects how alleviating chemotherapy has also become important problem.The health product that research and development control DNA toxic chemical endangers the mankind, have important society and economic worth.
Folium camelliae assamicae shines Folium Camelliae sinensis with the large leaf in original producton location, Yunnan to process, and is the tea beverage that people commonly use.It is fragrant that Folium camelliae assamicae has taste warm in nature, separates greasy cattle and sheep poison, pained by expectorant, and postcibal diarrhea such as to be removed heat by catharsis at the effect.Modern biomedical research also shows, Folium camelliae assamicae has the effects such as blood sugar lowering, blood fat reducing, blood pressure lowering, antiinflammatory, the health promoting beverage making Folium camelliae assamicae become people to like.But up to now, Folium camelliae assamicae for preventing the biohazard of DNA toxic chemical not yet to find, report by related data.
Summary of the invention
The object of the present invention is to provide a kind of compositions based on Folium camelliae assamicae main component, the constituent of said composition and mass percent thereof are: 5-50% abrownin, 5-40% tea polysaccharide, 10-50% tea polyphenols, 5-30% caffeine, 0.1-2% γ-aminobutyric acid and 0.1-2% gallic acid.
Another object of the present invention is to provide a kind of compositions novelty teabag based on Folium camelliae assamicae main component, namely using the compositions based on Folium camelliae assamicae main component as effective ingredient, and the application in the medicine of the biohazard of preparation prevention DNA poisonous chemical substance.
Compositions based on Folium camelliae assamicae main component is applied in preparation and strengthens chemotherapeutics to the lethal effect of tumor cell; strengthen tumor cell to the medicine of chemotherapy drug susceptibility, and for the preparation of protection chemotherapeutics to the application in the Normocellular toxic side effects of tumour patient.
Compositions based on Folium camelliae assamicae main component of the present invention is as the main active of its novelty teabag, further compositions can be made the derived products such as extractum, pill, oral liquid, tablet, injection, skin care implement, wherein can add one or more pharmaceutically acceptable adjuvants, to improve the assimilation effect of derived product or to be convenient to take.
Another object of the present invention is in the health product or the article of everyday use compositions based on Folium camelliae assamicae main component being applied in the biohazard of preparation prevention DNA poisonous chemical substance, namely using the present composition as main effectively active component, compositions is made health product or article of everyday use, add acceptable adjuvant in some health product or article of everyday use if desired, for preventing DNA poisonous chemical substance to Normocellular toxicity in daily life.
The present invention utilizes mouse embryo fibroblasts (the mouse embryo fibroblast of wild type; MEF) test dna toxic chemical is to Normocellular lethal effect; test compositions based on Folium camelliae assamicae main component to Normocellular protective effect simultaneously, thus disclose this compositions effect in the biohazard of prevention DNA toxic chemical.
The present invention utilizes Annexin V-FITC cell apoptosis detection kit to detect the apoptosis of cell and the protective effect of the present composition after the process of DNA poisonous chemical substance;
The principle detecting institute's foundation is as follows: Annexin V selective binding phosphatidyl serine, phosphatidyl serine is mainly distributed in inside cell membrane, namely adjacent with cytoplasm side, early stage at apoptosis, dissimilar cell all can translate into cell surface outside phosphatidyl serine, namely outside cell membrane; Blood coagulation and inflammatory reaction can be promoted after phosphatidyl serine is exposed to cell surface; And Annexin V and the phosphatidyl serine of translating into cell surface outward can block the coagulant blood of phosphatidyl serine after combining and proinflammatory is active; With the Annexin V of the fluorescent probe FITC labelling with green fluorescence, i.e. AnnexinV-FITC, this apoptotic key character that turns up of phosphatidyl serine just very simply and directly can be detected with flow cytometer or fluorescence microscope, after Annexin V-FITC and propidium iodide stain, normal living cells is not by AnnexinV-FITC and propidium iodide stain; The early stage cell of apoptosis is only dyeed by Annexin V-FITC, and propidium iodide stain is negative; The cell in non-viable non-apoptotic cell and apoptosis late period can simultaneously by Annexin V-FITC and propidium iodide stain.
Apoptosis detects and shows, part cell Annexin V after amycin process dyes as positive, and show that amycin starts apoptotic program, amycin (Doxorubicin, Dox) is toxic to normal cell; And add the cell that the present composition carries out protecting in advance, after amycin process, Annexin V dyes as positive cell quantity obviously reduces, and shows that the present composition has good protective effect to the Normocellular apoptosis that amycin is induced.
The present invention by immune-blotting method DNA damage stress associated protein p53, HSP70 expression, experimental result shows, after the process of DNA toxic chemical, in normal cell, the albumen such as p53, HSP70 can raise by response expression; And under the protection of the present composition, reduce the stress of DNA toxic chemical induction, show the DNA damage that the present composition alleviates toxic chemical and causes.
The present invention utilizes tumor cell p53
-/-+ S+Ras test dna toxic chemical is to the lethal effect of tumor cell, test the present composition to this sensitization killed and wounded simultaneously, thus disclose this compositions effect in clinical adjuvant chemotherapy, experimental data shows, after the process of DNA toxic chemical, Annexin V coloration result shows, after use DNA toxic chemical process tumor cell, apoptosis appears in cell; Use the present composition and DNA toxic chemical co-treatment, the apoptosis rate of tumor cell can be made obviously to increase, and this result shows that the present composition can strengthen the sensitivity of tumor cell to conventional chemotherapeutic drugs, can be applied to clinical adjuvant chemotherapy.
Therefore, experimentation of the present invention shows, the present composition has good protective effect to the Normocellular toxicity that chemotherapeutics causes, and tumor cell is then had to the auxiliaring effect of the lethal effect strengthening chemotherapeutics.This characteristic of the present composition may be used for clinical adjuvant chemotherapy: the normal cell protecting patient in chemotherapy process, alleviates toxicity, also can strengthen the lethal effect of chemotherapeutics to tumor cell simultaneously.This result imply that present composition adjuvant chemotherapy on clinical medicine, and reducing toxic and side effects aspect has good application prospect.In addition, the present composition in daily life for the preparation of eliminating the health product of DNA toxic chemical biohazard, also there is good application prospect article of everyday use aspect.
Compared with prior art, the present invention has following beneficial effect:
1, the present invention relates to the research and apply of sanipratics, toxicological chemistry, oncology, cell biology, propose first to apply the biohazard based on caused by the composition for preventing and controlling DNA toxic chemical of Folium camelliae assamicae main component, for Folium camelliae assamicae provides scientific basis for preventing and treating the biohazard of DNA toxic chemical.
2, the Normal cell death that causes after can preventing being exposed to DNA toxic chemical of the present composition, avoids its bio-toxicity; Preparation is used for research and develop anti-DNA damnification health product, the health product of the harm that prevents to smoke, neoadjuvant chemotherapy in clinical medicine, daily use chemicals industry etc.
3, provide the prevention and controls of economical and practical, simple DNA toxic chemical, disclose the novelty teabag of Folium camelliae assamicae, open up a new application.
4, economical and practical, simple chemotherapeutics photosensitivity-enhancing method is provided, for clinical tumor chemotherapy raising effect, reduction toxic and side effects provide new strategy.
Accompanying drawing explanation
Fig. 1 is normal apoptosis situation fluorescence staining schematic diagram after the process of DNA toxic chemical amycin; Wherein A is that normal cell dyes through amycin process 12 hours later cell Annexin V-FITC; B is normal cell through amycin process propidium iodide stain after 12 hours; C is the cellular morphology schematic diagram that differential interference micro-imaging (DIC) is observed;
Fig. 2 is when adding present composition protection, the situation schematic diagram of normal apoptosis after the process of DNA toxic chemical amycin; Wherein A is that under present composition protection, normal cell dyes through amycin process 12 hours later cell AnnexinV-FITC; B is under compositions protection, normal cell through amycin process propidium iodide stain after 12 hours; C is the cellular morphology that differential interference micro-imaging is observed;
Fig. 3 quantitatively adds before and after the present composition, and amycin causes the change schematic diagram of normal apoptosis rate; In figure, Dox is that the amycin not adding compositions causes Normocellular apoptosis rate; Dox+Pure is that after adding the present composition, amycin causes Normocellular apoptosis rate;
Fig. 4 be the present invention from molecular level, detect normal cell stress response protein expression schematic diagram; In figure, WT is the normal cell controls (wild type) without any process; WT+Pure is that normal cell adds present composition process; WT+Pure+Dox0.1 is after normal cell adds present composition process protection, then adds the DNA toxic chemical amycin process that dosage is 0.1ug/ml; WT+Dox0.1 is the DNA toxic chemical amycin process that normal cell directly adds that dosage is 0.1ug/ml; WT+Pure+Dox0.5 is after normal cell adds present composition process protection, then adds the DNA toxic chemical amycin process that dosage is 0.5ug/ml; WT+Dox0.5 is the DNA toxic chemical amycin process that normal cell directly adds that dosage is 0.5ug/ml;
Fig. 5 is the situation schematic diagram of apoptosis of tumor cells after the process of DNA toxic chemical amycin; Wherein A is that tumor cell 12 hours later cell Annexin V-FITC after amycin process dye; B is tumor cell propidium iodide stain after 12 hours after amycin process; C is the cellular morphology that differential interference micro-imaging is observed;
Fig. 6 is when adding the present composition, the situation schematic diagram of apoptosis of tumor cells after the process of DNA toxic chemical amycin; Wherein A is under present composition effect, and tumor cell dyes through amycin process 12 hours later cell AnnexinV-FITC; B is under present composition effect, and tumor cell is through amycin process propidium iodide stain after 12 hours; C is the cellular morphology that differential interference micro-imaging is observed;
Fig. 7 is that Quantitative Comparison is added before and after the present composition, and amycin causes the change schematic diagram of apoptosis of tumor cells rate; In figure, Dox is the apoptosis rate that the amycin not adding the present composition causes tumor cell; Dox+Pure is the apoptosis rate that after adding the present composition, amycin causes tumor cell;
Fig. 8 is that MTT detection by quantitative is added before and after the present composition, and cyclophosphamide causes the change of death of neoplastic cells degree; In figure, abscissa is concentration of cyclophosphamide, and vertical coordinate is cell survival degree, and puer is the test group of adding the present composition, and control is the matched group not adding the present composition;
Fig. 9 is that MTT detection by quantitative is added before and after the present composition, and carboplatin causes the change of death of neoplastic cells degree; In figure, abscissa is carboplatin concentration, and vertical coordinate is cell survival degree, and puer is the test group of adding the present composition, and control is the matched group not adding the present composition.
Detailed description of the invention
Below in conjunction with drawings and Examples, the present invention is described in further detail; but scope is not limited to described content; the method used in the present embodiment is conventional method if no special instructions, uses reagent if no special instructions in the present embodiment, is commercially available conventional reagent.
Embodiment 1: based on the compositions of Folium camelliae assamicae main component in the present invention, its constituent and mass percent are 50% abrownin, 30% tea polysaccharide, 11% tea polyphenols, 5% caffeine, 2% γ-aminobutyric acid and 2% gallic acid.
The present embodiment utilizes mouse embryo fibroblasts (the mouse embryo fibroblast of wild type, MEF) test dna toxic chemical is to Normocellular lethal effect, test the present composition to Normocellular protective effect simultaneously, thus disclose this compositions effect in the biohazard of prevention DNA toxic chemical;
The present embodiment utilizes Annexin V-FITC cell apoptosis detection kit to detect the protective effect of apoptosis and compositions, and concrete operations are as follows: inoculum density is 1 × 10
5individual mouse embryo fibroblasts, in the glass slide in six porocyte culture plates, uses DMEM+10% hyclone culture medium, at 37 DEG C, 5%CO
2, 3%O
2cell attachment is cultured in incubator; Cell is divided into test group and matched group, the present embodiment compositions Dual culture is added in test group, matched group does not add the present embodiment compositions Dual culture, cultivate after 12 hours, the amycin adding 0.1 μ g/mL or 0.5 μ g/mL in the present embodiment compositions test group cell and cellular control unit respectively processes, after process 12h, carry out Annexin V fluorescence staining to cell, dyeing concrete steps are as follows:
1, absorb the cell culture fluid in Tissue Culture Plate glass slide, add PBS buffer solution once;
2,195 μ l Annexin V-FITC are added in conjunction with liquid;
3, add 5 μ l Annexin V-FITC, mix gently, room temperature 25 DEG C of lucifuges hatch 10 minutes;
4, absorb dyeing liquor, add 190 μ l Annexin V-FITC in conjunction with liquid;
5, add 10 μ l propidium iodide stain liquid, mix gently, room temperature lucifuge is placed;
Under fluorescence microscope, carry out Annexin V Fluorescent Staining Observation, Annexin V-FITC is green fluorescence, and propidium iodide is red fluorescence, auxiliary with the metamorphosis of differential interference imaging microscopy cell.
The results are shown in Figure 1-4, in Fig. 1-A figure, fluorescence staining bright spot is the positive cell that early apoptosis occurs after 12 hours normal cell after amycin process; In B figure, fluorescence staining bright spot is the cell that late apoptic or necrosis occur after 12 hours normal cell after amycin process;
In Fig. 2-A figure, fluorescence staining bright spot is that under the protection of the present embodiment compositions, the positive cell of early apoptosis occurs after 12 hours through amycin process normal cell; In B figure, fluorescence staining bright spot is that under compositions protection, normal cell, through amycin process, the cell of late apoptic or necrosis occurs for 12 hours;
Fig. 3 is the quantitative result to Fig. 1-2, and after display the present embodiment compositions makes amycin process, normal apoptosis rate is reduced to 4% from 18%, shows good protected effect;
As can be seen from the above results: cell Annexin V after amycin process of about 18% dyes as positive, and show that amycin starts Normocellular apoptosis program, amycin (Doxorubicin, Dox) is toxic to normal cell; And adding the cell that the present embodiment compositions carries out protecting in advance, the 4%Annexin V that only has an appointment after the process of interpolation amycin dyes as positive, shows that the present composition has good protective effect to the Normocellular apoptosis that amycin is induced.
Fig. 4 be from molecular level by immune-blotting method DNA damage stress associated protein p53, HSP70 expression.Experimental result shows, normal MEF cell is after the amycin process of 0.1 μ g/mL or 0.5 μ g/mL, and in cell, the albumen such as p53, HSP70 can raise (WT+Dox0.1, WT+Dox0.5) by response expression; And under the protection of the present embodiment compositions; reduce the stress (WT+Pure+Dox0.1 of the albumen such as p53, HSP70 of DNA toxic chemical amycin induction; WT+Pure+Dox0.5), the DNA damage that the present embodiment compositions alleviates toxic chemical amycin and causes is shown.
Embodiment 2: based on the compositions of Folium camelliae assamicae main component in the present invention, its constituent and mass percent are 5% abrownin, 40% tea polysaccharide, 30% tea polyphenols, 23% caffeine, 1% γ-aminobutyric acid and 1% gallic acid.
The present embodiment utilizes tumor cell p53
-/-+ S+Ras test dna toxic chemical is to the lethal effect of tumor cell, and test the present embodiment compositions to this sensitization killed and wounded simultaneously, thus disclose this compositions effect in clinical adjuvant chemotherapy, concrete operations are as follows: inoculum density is 1 × 10
5individual tumor cell p53
-/-+ S+Ras, in the glass slide in six porocyte culture plates, uses DMEM+10% hyclone culture medium, at 37 DEG C, 5%CO
2, 3%O
2cell attachment is cultured in incubator; Cell is divided into test group and matched group, the present embodiment compositions Dual culture is added in test group, matched group does not add the present embodiment compositions Dual culture, cultivate after 12 hours, amycin the present embodiment compositions test group cell and cellular control unit being added respectively to 0.1 μ g/mL or 0.5 μ g/mL processes, after process 12h, carry out Annexin V fluorescence staining to cell, staining procedure is as follows:
1, absorb the cell culture fluid in Tissue Culture Plate glass slide, add PBS buffer solution once;
2,195 μ l Annexin V-FITC are added in conjunction with liquid;
3, add 5 μ l Annexin V-FITC, mix gently, room temperature 20 DEG C of lucifuges hatch 10 minutes;
4, absorb dyeing liquor, add 190 μ l Annexin V-FITC in conjunction with liquid;
5, add 10 μ l propidium iodide stain liquid, mix gently, room temperature lucifuge is placed.
Under fluorescence microscope, carry out Annexin V Fluorescent Staining Observation, Annexin V-FITC is green fluorescence, and propidium iodide is red fluorescence, auxiliary with the metamorphosis of differential interference imaging microscopy cell.
The results are shown in Figure 5-7, in Fig. 5-A, fluorescence staining bright spot is the positive cell that early apoptosis occurs after 12 hours tumor cell after amycin process; In B figure, fluorescence staining bright spot is the cell that late apoptic or necrosis occur after 12 hours tumor cell after amycin process;
For under fluorescence staining bright spot is the effect of the present embodiment compositions in Fig. 6-A, there is the positive cell of early apoptosis in tumor cell after 12 hours through amycin process; In B figure, fluorescence staining bright spot is under the effect of the present embodiment compositions, and the cell of late apoptic or necrosis occurs after 12 hours through amycin process tumor cell;
Fig. 7 is the quantitative result to Fig. 5-6, show the present embodiment compositions make amycin process after apoptosis of tumor cells rate be increased to 22% from 12%, show the good Chemotherapy of compositions, auxiliary effect of killing and wounding;
As can be seen from the above results: after the process of DNA toxic chemical, Annexin V coloration result shows, there is apoptosis in tumor cell about 12% cell, uses the present embodiment compositions and DNA toxic chemical co-treatment, the apoptosis rate of tumor cell can be made to rise to 22%; This result shows that this example composition can strengthen the sensitivity of tumor cell to conventional chemotherapeutic drugs, can be applied to clinical adjuvant chemotherapy.
Embodiment 3: based on the compositions of Folium camelliae assamicae main component in the present invention, its constituent and mass percent are 14.8% abrownin, 5% tea polysaccharide, 50% tea polyphenols, 30% caffeine, 0.1% γ-aminobutyric acid and 0.1% gallic acid.
The present embodiment, using cyclophosphamide as DNA toxic chemical, utilizes tumor cell p53
-/-+ S+Ras test dna toxic chemical is to the lethal effect of tumor cell, and test the present embodiment compositions to this sensitization killed and wounded simultaneously, thus disclose this compositions effect in clinical adjuvant chemotherapy, concrete operations are as follows: inoculation 5 × 10
3individual tumor cell p53
-/-+ S+Ras is in 96 porocyte culture plates, and use DMEM+10%FBS culture medium, detection sample is divided into test group and matched group, adds the present embodiment compositions Dual culture in test group, matched group does not add the present embodiment compositions Dual culture, at 37 DEG C, and 5%CO
2, 3%O
2cultivate in incubator; cultivate 24 hours respectively, the cyclophosphamide then adding 0.05,0.1,0.5,1,5 μ g/mL respectively at the present embodiment compositions protection group cell and cellular control unit processes, after continuing to cultivate 48h; carry out MTT detection by quantitative, it is as follows that MTT detects concrete steps:
1, every hole adds 20ulMTT solution (5mg/ml, i.e. 0.5%MTT), continues cultivation 4 hours;
2, stop cultivating, carefully suck culture fluid in hole, add PBS buffer solution once;
3, every hole adds 150ul dimethyl sulfoxide, puts low-speed oscillation 5min on shaking table, crystal is fully dissolved; The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument OD490nm place.
The results are shown in Figure 8, MTT detection by quantitative shows, after adding the present composition, cyclophosphamide causes tumor cell survival degree than the matched group (control) not adding the present composition obviously to be reduced, also i.e. dead degree increase more obvious than matched group, shows that the present composition can enhanced sensitivity cyclophosphamide killing and wounding tumor cell.
Embodiment 4: based on the compositions of Folium camelliae assamicae main component in the present invention, its constituent and mass percent are 30% abrownin, 39% tea polysaccharide, 10% tea polyphenols, 20% caffeine, 0.5% γ-aminobutyric acid and 0.5% gallic acid.
The present embodiment, using carboplatin as DNA toxic chemical, utilizes tumor cell p53
-/-+ S+Ras test dna toxic chemical is to the lethal effect of tumor cell, and test the present embodiment compositions to this sensitization killed and wounded simultaneously, thus disclose this compositions effect in clinical adjuvant chemotherapy, concrete operations are as follows: inoculation 5 × 10
3individual tumor cell p53
-/-+ S+Ras is in 96 porocyte culture plates, and use DMEM+10%FBS culture medium, detection sample is divided into test group and matched group, adds the present embodiment compositions Dual culture in test group, matched group does not add the present embodiment compositions Dual culture, at 37 DEG C, and 5%CO
2, 3%O
2cultivate in incubator, cultivate 24 hours respectively, the carboplatin then adding 0.05,0.1,0.5,1,5 μ g/mL respectively at the present embodiment compositions protection group cell and cellular control unit processes.After continuing to cultivate 48h, carry out MTT detection by quantitative, MTT detecting step is as follows:
1, every hole adds 20ulMTT solution (5mg/ml, i.e. 0.5%MTT), continues cultivation 4 hours;
2, stop cultivating, carefully suck culture fluid in hole, add PBS washing once;
3, every hole adds 150ul dimethyl sulfoxide, puts low-speed oscillation 5min on shaking table, crystal is fully dissolved, and measures the light absorption value in each hole at enzyme-linked immunosorbent assay instrument OD490nm place.
The results are shown in Figure 9, MTT detection by quantitative to show, add after the present composition, carboplatin causes tumor cell survival degree than the matched group (control) not adding the present composition obviously to be reduced, and is also the increase more obvious than matched group of dead degree.Show that the present composition can enhanced sensitivity carboplatin killing and wounding tumor cell.
Embodiment 5: based on the compositions of Folium camelliae assamicae main component in the present invention, its constituent and mass percent are 50% abrownin, 30% tea polysaccharide, 11% tea polyphenols, 5% caffeine, 2% γ-aminobutyric acid and 2% gallic acid.
The present embodiment utilizes the mouse embryo fibroblasts test dna toxic chemical cyclophosphamide of wild type to Normocellular lethal effect, test the present composition to Normocellular protective effect simultaneously, thus disclose this compositions and alleviating the effect in cyclophosphamide toxic and side effects;
Concrete operations are as follows: inoculum density is 1 × 10
5individual mouse embryo fibroblasts, in the glass slide in six porocyte culture plates, uses DMEM+10% hyclone culture medium, at 37 DEG C, 5%CO
2, 3%O
2cell attachment is cultured in incubator; Cell is divided into test group and matched group, the present embodiment compositions Dual culture is added in test group, matched group does not add the present embodiment compositions Dual culture, cultivate after 12 hours, the cyclophosphamide adding 0.5 μ g/mL in the present embodiment compositions test group cell and cellular control unit respectively processes, after process 12h, cell is observed; Experimental result shows: before the protection of the present embodiment compositions, cell mortality is 20%; after the protection of interpolation the present embodiment compositions, cell mortality is 8%; test group cell is compared with cellular control unit; floating number of dead cells obviously reduces, and shows that the present embodiment compositions can alleviate cyclophosphamide to Normocellular lethal effect.
Embodiment 6: based on the compositions of Folium camelliae assamicae main component in the present invention, its constituent and mass percent are 30% abrownin, 39% tea polysaccharide, 10% tea polyphenols, 20% caffeine, 0.5% γ-aminobutyric acid and 0.5% gallic acid.
The present embodiment utilizes the mouse embryo fibroblasts test dna toxic chemical carboplatin of wild type to Normocellular lethal effect, test the present composition to Normocellular protective effect simultaneously, thus disclose this compositions and alleviating the effect in carboplatin toxic and side effects;
Concrete operations are as follows: inoculum density is 1 × 10
5individual mouse embryo fibroblasts, in the glass slide in six porocyte culture plates, uses DMEM+10% hyclone culture medium, at 37 DEG C, 5%CO
2, 3%O
2cell attachment is cultured in incubator, cell is divided into test group and matched group, the present embodiment compositions Dual culture is added in test group, matched group does not add the present embodiment compositions Dual culture, cultivate after 12 hours, the carboplatin adding 0.5 μ g/mL in the present embodiment compositions test group cell and cellular control unit respectively processes, after process 12h, cell is observed, experimental result shows: before the protection of the present embodiment compositions, cell mortality is 10%, after the protection of interpolation the present embodiment compositions, cell mortality is 5%, test group cell is compared with cellular control unit, floating number of dead cells obviously reduces, show that the present embodiment compositions can alleviate carboplatin to Normocellular lethal effect.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention.All any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (2)
1. the compositions based on Folium camelliae assamicae main component strengthens chemotherapeutics to a lethal effect for tumor cell as sole active agent in preparation, the application in the medicine simultaneously protecting normal cell to kill and wound from chemotherapeutics;
Constituent and the mass percent of the described compositions based on Folium camelliae assamicae main component are: 5-50% abrownin, 5-40% tea polysaccharide, 10-50% tea polyphenols, 5-30% caffeine, 0.1-2% γ-aminobutyric acid and 0.1-2% gallic acid.
2. according to claim 1 based on the application of the compositions of Folium camelliae assamicae main component, it is characterized in that: compositions is extractum, pill, oral liquid, tablet or injection.
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CN106177187B (en) * | 2016-07-26 | 2020-02-21 | 天津大学 | Tea polyphenol tea polysaccharide composition with synergistic attenuation and anti-liver cancer effects |
CN111840412B (en) * | 2020-08-21 | 2022-03-08 | 杭州茗褐生物科技有限公司 | Application of theabrownin in preparation of anti-melanoma drugs |
CN112957406A (en) * | 2021-04-20 | 2021-06-15 | 吉林大学 | Application of cooked puerh tea aqueous extract in preparation of medicine for enhancing organism immunity and/or inflammation level |
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