CN104689088B - A kind of Chinese medicine preparation and preparation method of prevention immunosenescence - Google Patents
A kind of Chinese medicine preparation and preparation method of prevention immunosenescence Download PDFInfo
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Abstract
The present invention provides a kind of Chinese medicine preparations of prevention immunosenescence, are formed by following parts by weight medicine preparation：Radix rehmanniae preparata 6 600, the stem of noble dendrobium 5~500, Chinese yam 5~500, matrimony vine 5~500, Fructus Corni 4~400, Semen Cuscutae 6~600, Cortex Eucommiae 4~400, Chinese cassia tree 3~300, Radix Angelicae Sinensis 3~300, Radix Astragali 6 600, Radix Paeoniae Alba 5 500, radix polygonati officinalis 5 500, Rhizoma Chuanxiong 5 500；Meanwhile the present invention also provides preparation methods；The present invention can target the activity for promoting Tn and Th1 class subgroups in T cell subgroup while improving the lymphoid organs indexes such as spleen, thymus gland and NK, NKT cell activity, can prevent T cell immunosenescence and promote T cell immunologic reconstitution after radiotherapy.
Description
Technical field
The present invention relates to the Chinese medicines of T cell and NK immune reconstitutions after a kind of prevention T cell immunosenescence, promotion radiotherapy
Compatibility agent is using Chinese medicine as pharmaceutical composition made of Raw material processing.Controllability is strong, and curative effect is clear.It can be used for radiation immunity
T cell immunologic reconstitution and the prevention of major disease etc. after damage, tumor radiotherapy.
Background technology
T cell is huge and height heterogeneity group, accounts for 60% of immune cell or so.It can be divided into surface marker
CD4+, CD8+T cell subsets, wherein T cells can be divided into according to differentiation state difference CD4+T cells（Naive T,
Tn）, memory t cell（Memory T cell, Tm）Not with effector T cell (Teff), effector T cell and according to functional effect
It is same to be divided into multiple T cell subgroups such as Th1/Th2/Treg/Th17.The classification of CD8 is identical as CD4+T cells.T cell diversity is
Protect the important regulation mechanism of homeostasis.T cells can be divided into Th1/Th2/Treg/Th17 etc. after meeting antigenic stimulus
Multiple subgroups：Th1 has the effects that antiviral, anticancer, Th1 disadvantages that can cause infectious diseases and tumour etc.；Th2 can promote antibody
It generates, has antibacterial, antitoxin effect, the diseases such as Th2 disadvantages and parasitic infection and immunological rejection related.It is right that Th1 and Th2 passes through
Vertical adjust limits mutual response intensity.In addition, Treg subgroups (TGF β+) can restricted T h1/Th2 hyperpolarizations.Treg is overexpressed
It is closely related with tumor development.In addition, Th17 is related to tissue inflammation or tumour inflammation, Th1/Th2 type cytokines can limit
The hyper-proliferative of the subgroup, recently research confirm：IFNg+IL-17+Th17 is also the anti-inflammatory antitumor potential of body.It removes
Except this, Tm cells are also the protrusion of immunosenescence with the repetitious stimulation for increasing age and antigen, in vivo a large amount of accumulations, Tm enrichments
Feature.But Tm is not good for nothing, part TEMAt immune response initial stage, rapid secretion IFNg is to start or instruct the innate immunity thin
The valuable source of born of the same parents NK cell clearances cause of disease and tumour cell.Therefore, T cell subgroup is unbalance, is especially reduced to Tn, Th1
The immunosenescence of main feature is that tumour, autoimmunity disease, great infectious diseases threaten the prominent features of the elder's health.Targeting
It is one of the important control measure for lowering major disease and threatening human health that T cell, which improves immunosenescence state,.Existing Chinese medicine is mended
Beneficial class drug is classified with class of enriching blood, tonifying Qi class, yang-tonifying class and tonifying yin class mostly, can not correspond to each subgroup of T cell in various disease
The different modern study achievement of middle advantage, cannot be directed to various disease and effectively play has the stronger treatment of cell subsets specific aim
Effect.Such as, the Tn with immune plasticity and Th1, Tc1 with antiviral and antitumor is that physical exertion resists infectious disease
Sick and antitumor important cells group, the inferior position of these cell subsets lead to infectious diseases and tumor development
Major reason.And the cell subsets such as Th2, Th17, Treg are unfavorable for the performance of antiviral and antitumor effect.Therefore, it improves immune
Drug should have preferable cell selective and effect specific aim.Currently, the Chinese medicine for adjusting cellular immunity has Zuogui Wan, kidney tonifying
The reports such as side, but its curative effect is only improved the effect report of Immune Organs Index, NK cells, T cell entirety, there is no targeting different
T cell subgroup plays cell selective and the effect stronger drug matching preparation of specific aim.
Invention content
In order to solve the above technical problems, the present invention provides a kind of Chinese medicine preparation of prevention immunosenescence, provide simultaneously
Preparation method.
The present invention can target while improving the lymphoid organs indexes such as spleen, thymus gland and NK, NKT cell activity
The activity for promoting Tn and Th1 class subgroups in T cell subgroup can prevent T cell immunosenescence and promote the immune weight of T cell after radiotherapy
It builds.
Technical scheme is as follows：A kind of Chinese medicine preparation of prevention immunosenescence, is formed by following medicine preparation：
Radix rehmanniae preparata 6-600, the stem of noble dendrobium 5~500, Chinese yam 5~500, matrimony vine 5~500, Fructus Corni 4~400, Semen Cuscutae 6~600,
Cortex Eucommiae 4~400, Chinese cassia tree 3~300, Radix Angelicae Sinensis 3~300, Radix Astragali 6-600, Radix Paeoniae Alba 5-500, radix polygonati officinalis 5-500, Rhizoma Chuanxiong 5-500.
The preparation method of the present invention is as follows：Above each taste mixing is taken, 10 times of 75% ethyl alcohol of volume, leaching are added after water rinses
2h is steeped, reflux decocts 1.5h, leaching liquid, and 10 times of 50% ethyl alcohol of volume are added in the dregs of a decoction, and reflux decocts 1.5h, and leaching liquid merges
Liquid, 80 DEG C are concentrated under reduced pressure recycling ethyl alcohol, collect alcohol extracting clear cream, obtain ethanol extract.10 times of volume of water are added in the dregs of a decoction, decoct
1.5h, leaching liquid is condensed into cream, merges ethyl alcohol and water extract-alcohol precipitation extract, paste, aqua, particle, syrup etc. is made, makes
1.92g/ml containing crude drug.
Pharmaceutical composition of the present invention contains water and extractive with organic solvent, contains polysaccharide, amino acid, cinnaldehydrum, ursolic acid, soap
The plurality of active ingredients such as glycosides, alkaloid, vitamin, trace element.Organic solvent is ethyl alcohol.
Curative effect of the present invention is clear, and has preferable cell selective, in clear Tn/Tem/Tcm and Th1/Th2/
After the specific variation characteristic of Treg/Th17 T effector cells subgroup and NK cells after radiation immune injury, targetedly improve
The proliferation and function of Tn/Tem, Th1 and NK cell subsets, selective depression inflammatory T cell subgroup Th17 and inhibitive ability of immunity T are thin
The proliferation and function factor of born of the same parents' subgroup Treg discharges, and in immunologic reconstitution after promoting T cell radiation, selectivity raising has anti-swollen
The proliferation of tumor, antiviral functional lymphocyte subgroup, advantage make antitumor microenvironment and the capacity building of body.
Description of the drawings
Fig. 1 is the apoptotic state figure for improving thymocyte；
Fig. 2 is the activation figure for promoting thymocyte；
Fig. 3 is the activation figure for promoting Tn and Tem lymphocytes；
Fig. 4 is that Th1 is promoted to inhibit Th17/Treg lymphocyte figures；
Fig. 5 is that the proliferation of NK cells is promoted to scheme with function factor release；
Fig. 6 is the Lung metastases figure of B16 tumour cells after inhibiting radiation.
Specific implementation mode
With reference to embodiment, the present invention is further illustrated.
Embodiment 1
Take radix rehmanniae preparata 6g, stem of noble dendrobium 5g, Chinese yam 5g, matrimony vine 5g, Fructus Corni 4g, Semen Cuscutae 6g, Cortex Eucommiae 4g, Chinese cassia tree 3g, Radix Angelicae Sinensis 3g,
10 times of 75% ethyl alcohol of volume are added in Radix Astragali 6g, Radix Paeoniae Alba 5g, radix polygonati officinalis 5g, Rhizoma Chuanxiong 5g after water rinses, and impregnate 2h, and reflux decocts
10 times of 50% ethyl alcohol of volume are added in 1.5h, leaching liquid, the dregs of a decoction, and reflux decocts 1.5h, and leaching liquid merges liquid, 80 DEG C of decompressions
Concentration and recovery ethyl alcohol collects alcohol extracting clear cream, obtains ethanol extract.10 times of volume of water are added in the dregs of a decoction, decoct 1.5h, leaching liquid,
It is condensed into cream, merges ethyl alcohol and water extract-alcohol precipitation extract, paste, aqua, particle, syrup etc. is made, makes 1.92g/ml containing crude drug.
Embodiment 2
Take radix rehmanniae preparata 400, the stem of noble dendrobium 500, Chinese yam 200, matrimony vine 300, Fructus Corni 400, Semen Cuscutae 400, Cortex Eucommiae 400, Chinese cassia tree
100,10 times of 75% ethyl alcohol of volume, leaching is added in Radix Angelicae Sinensis 300, Radix Astragali 600, Radix Paeoniae Alba 500, radix polygonati officinalis 500, Rhizoma Chuanxiong 300 after water rinses
2h is steeped, reflux decocts 1.5h, leaching liquid, and 10 times of 50% ethyl alcohol of volume are added in the dregs of a decoction, and reflux decocts 1.5h, and leaching liquid merges
Liquid, 80 DEG C are concentrated under reduced pressure recycling ethyl alcohol, collect alcohol extracting clear cream, obtain ethanol extract.10 times of volume of water are added in the dregs of a decoction, decoct
1.5h, leaching liquid is condensed into cream, merges ethyl alcohol and water extract-alcohol precipitation extract, paste, aqua, particle, syrup etc. is made, makes
1.92g/ml containing crude drug.
Embodiment 3
Take radix rehmanniae preparata 600, the stem of noble dendrobium 400, Chinese yam 500, matrimony vine 400, Fructus Corni 100, Semen Cuscutae 600, Cortex Eucommiae 150, Chinese cassia tree
300,10 times of 75% ethyl alcohol of volume, leaching is added in Radix Angelicae Sinensis 100, Radix Astragali 400, Radix Paeoniae Alba 200, radix polygonati officinalis 200, Rhizoma Chuanxiong 500 after water rinses
2h is steeped, reflux decocts 1.5h, leaching liquid, and 10 times of 50% ethyl alcohol of volume are added in the dregs of a decoction, and reflux decocts 1.5h, and leaching liquid merges
Liquid, 80 DEG C are concentrated under reduced pressure recycling ethyl alcohol, collect alcohol extracting clear cream, obtain ethanol extract.10 times of volume of water are added in the dregs of a decoction, decoct
1.5h, leaching liquid is condensed into cream, merges ethyl alcohol and water extract-alcohol precipitation extract, paste, aqua, particle, syrup etc. is made, makes
1.92g/ml containing crude drug.
The test of pesticide effectiveness
Method：Using C57BL/6 inbred mouses respectively through X-ray（2.8Gy, dosage rate are 1.35Gy min-1, away from
From 170cm）Radiation immune injury model is established in irradiation, and using the methods of flow cytometry and RT-PCR, observation radiation is to thymus gland
The influence of development and cell differentiation and mature T cells subpopulations and its protective effect of the present invention.It is withered by detecting thymocyte
Die state, the expression of apoptosis-related genes (bcl2/bax, caspases), specific phenotypes' molecular composition（CD4+SP,CD8+SP,
CD4+CD8+DP）, thymus dendritic cells phenotypic characteristic and ceS signal molecules (LFA-1, ICAM-1), periphery device is immunized
The subgroups such as official Tn/TEM/TCM, Th1/Th2/Th17/Treg and its cytokine secretion patterns and the variation shape of reply radiation
State, the unkehr effect etc. of the present invention.
（1）X-ray irradiation mouse model：Health C57BL/6 inbred mouses 21 are selected, are randomly divided into 3 groups（Every group 7
Only）：Negative control group and irradiation model group mouse freely ingest drinking-water；Experimental group takes the embodiment of the present invention 1 and prepares drug,
After continuously drinking 10 days, using X-ray full-body exposure, distance 170cm. accumulated doses are 3.0Gy；Continue after taking 7~10 days,
Each group mouse is put to death, the immune organs mononuclearcell such as separating mouse peripheral blood, spleen, marrow, thymus gland detects Tn, TCM、TEM、
The T cells subgroup Proliferation, Differentiation state such as Th1, Th2, Th17, Treg.
（2）Tumor model mouse：Using healthy C57BL/6 inbred mouses 21, it is randomly divided into 3 groups（Every group 7）：It is cloudy
Property control group and irradiation model mouse freely ingest drinking-water；Experimental group of the present invention after continuously drinking 10 days, is shone using X-ray whole body
It penetrates, distance 170cm. accumulated doses are 3.0Gy；Irradiate third day, mouse tail vein injection B16 mouse melanin tumor cells（2×
105/ 300 μ l/ are only）, continue continuously to take Chinese medicine preparation, put to death mouse after 10 days according to a conventional method, detect lungs tumor burden
Amount（Tumor colonies number）, the immune organs mononuclearcell such as separating mouse peripheral blood, spleen, marrow, thymus gland, detection Tn, TCM、
TEM, the T cells subgroup Proliferation, Differentiation state such as Th1, Th2, Th17, Treg.
As a result：
1. protective effect of the invention formulation to radiation patterns thymocyte apoptosis：
Host cell group of the T cell as immune system is the leading role during radiation-induced overall immunity declines.Thymus gland
It is one of the important measures for protecting immunity of organism strength as the energy reserves maincenter for expanding periphery T cell library.Experimental group
（OCT）Thymocyte apoptosis rate can be reduced, thymic stromal cell antigen presenting molecule is improved（I-Ad）Ability to express, promote
CD4+CD8+DP is activated, and improves the viability of thymus gland DP cells, is enhanced intercellular information and is transmitted, be T cell differentiation and development and
Maturation provides microenvironment guarantee.As a result as depicted in figs. 1 and 2.
To the adjustment effect feature of Tn/Tm cellular reconstitutions after radiation：
Experimental synchronous observes experimental group Chinese medicine preparation and is directed to lymphoid organ T cell subgroup Tn/TEM/TCM, finds experiment
Group can effectively protect Tn maturations differentiation, improves the distribution capability in lymphoid organ, resource is provided for the activation of T effector cell.
T cell immunologic reconstitution phase after irradiation, TEMIt is the faster a group cell of reconstruction ability, OCT has selection to this cell subset
Property facilitation, and to the T with immunologic inertiaCMPlay opposite inhibiting effect (result is shown in Fig. 3).
Optimize influence of the agent to T effector cell subset cytokine secretion capacity during immunologic reconstitution：It is damaged in radiation
After wound, Th2 class cell subsets does not significantly change, and participates in antitumor, antiviral Th1 type cytokines and be remarkably decreased, this
It is the major reason of T cell function reduction after radiating.The ability that experimental group OCT secretes radiation-induced T cell IFNg declines
With significant protective effect（Fig. 4 A）.Ratio and quantity obviously rise Th17 cell subsets during immunologic reconstitution after irradiation
Height, OCT have restriction effect to the reconstruction of this group of cells（P<0.05, Fig. 4 B）.Treg cell subsets is immunized after radiation injury
Ratio is apparently higher than normal control in reconstruction process, and OCT Chinese medicine preparations can lower the proliferation of Treg（P<0.05, Fig. 4 C）.
Promote the proliferation of NK cells and activation after radiating
NK cells play as the main cell group of the innate immunity in antitumor, antiviral immunity reaction process important
Role, there are some researches prove the activation of T cell subgroup especially Tem can promote the proliferation and function of NK cells.OCT preparation exists
When radiation murine Tem being promoted to rebuild, how generated on NK cells influences and one of critical issue that we inquire into.Research hair
It is existing, although NK ratios do not have significant changes (Fig. 5 A), the ability of NK secretions IFNg to be decreased obviously after irradiation.As OCT promotes Tem
Reparation, OCT processing group NK cell proportions are significantly raised, and the ability of NK secretions IFNg be remarkably reinforced (P<0.01 Fig. 5 B).
Protect the anti-tumor effect of radio-immunity damage
The above results show that OCT Chinese medicine preparations can advantage improve radiation after immunosenescence mouse Tn, TEM and responsiveness T
The immunologic reconstitution of the cell subsets such as cell Th1 and NK, the advantage repair ability of these cell subsets, if after radiation capable of being improved
Immunosenescence mouse resists the effect of metastases, further to verify this as a result, we are that mouse tail vein is noted after radiating
B16 melanoma cells, and synchronous feeding OCT preparation are penetrated, it is found that mouse after said preparation is handled, there is obviously x-ray bombardment
Protective effect, lungs metastases clone significantly reduce, such as Fig. 6.
Conclusion：T cell expands the initial T of peripheral immune organ after radiation in the differentiation and development of thymus gland after OCT promotes radiation
Cell content supplements the T cell subpopulations energy, and has Balanceregulation to the differentiation and dysfunction of T cell subgroup：
Alternative promotes the immunologic reconstitution of the subgroups and NK cells such as Tn/TEM, Th1, and it is antitumor, antiviral, anti-infective to be conducive to body
Immunologic reconstitution, this with result can make said preparation become new immunopotentiator, be clinically used for the immune protection of radiotherapy and exempt from
Epidemic disease rebuilds accelerating agent, and the immunocompromiseds relevant disease such as prevention metastases has important clinical value.
Claims (2)
1. a kind of Chinese medicine composition of prevention immunosenescence, which is characterized in that formed by following parts by weight medicine preparation：Radix rehmanniae preparata
400-600, the stem of noble dendrobium 400~500, Chinese yam 200~500, matrimony vine 300~500, Fructus Corni 100~400, Semen Cuscutae 400~600,
Cortex Eucommiae 150~400, Chinese cassia tree 100~300, Radix Angelicae Sinensis 100~300, Radix Astragali 400-600, Radix Paeoniae Alba 200-500, radix polygonati officinalis 200-500, river
Rhizome of chuanxiong 300-500.
2. the preparation method of Chinese medicine composition according to claim 1, which is characterized in that include the following steps：It takes above
Each taste mixing, is added 10 times of 75% ethyl alcohol of volume after water rinses, and impregnates 2h, and reflux decocts 1.5h, leaching liquid, and the dregs of a decoction are added
10 times of 50% ethyl alcohol of volume, reflux decoct 1.5h, and leaching liquid merges liquid, and 80 DEG C are concentrated under reduced pressure recycling ethyl alcohol, collect alcohol extracting
Clear cream obtains ethanol extract；10 times of volume of water are added in the dregs of a decoction, decoct 1.5h, and leaching liquid is condensed into cream, merges ethyl alcohol and water
Paste, aqua, particle, syrup is made in extract, makes 1.92g/ml containing crude drug.
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| CN101721590A (en) * | 2008-10-10 | 2010-06-09 | 刘茂松 | Formula of capsules for effectively improving activity of life cells and production process method of capsules |
| CN103834537A (en) * | 2012-11-24 | 2014-06-04 | 高炳良 | Qi-tonifying wine |
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