CN103565901A - Health-care compound and preparation method thereof - Google Patents
Health-care compound and preparation method thereof Download PDFInfo
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- CN103565901A CN103565901A CN201310491336.8A CN201310491336A CN103565901A CN 103565901 A CN103565901 A CN 103565901A CN 201310491336 A CN201310491336 A CN 201310491336A CN 103565901 A CN103565901 A CN 103565901A
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- stem cell
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- herb gynostemmae
- gynostemmae pentaphylli
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Abstract
The invention discloses a bioactive substance composition and a preparation method thereof. The method comprises the following steps: firstly, extracting a healthy humanized stem cell, collecting the stem cell in a specific stage through in vitro culture and amplification, and extracting the bioactive substance of the stem cell at a low temperature; secondly, respectively extracting gynostemma pentaphylla and germination activated ganoderma spore bioactive substances according to corresponding methods; thirdly, combining the three bioactive substances according to a certain proportion to form the composition which is enriched with stem cell factor population, polypeptide, natural immunoprotein, nucleic acid, saponin, flavonoids, polysaccharide, amino acid, triterpenoid, alkaloid, ganoderma lucidum sterol, active enzyme and other active substances. The composition can be developed into products with cytobiological active functions, and has the effects of activating stem cells of a human body, repairing or replacing damaged, lesion and aged cells, adjusting blood fat, blood sugar and blood pressure, strengthening physical immunity, resisting tumor and aging and the like after acting in a human body. The composition has an extreme application prospect in medicine health-care and beautifying fields.
Description
Technical field
The invention belongs to biological technical field, be specifically related to: 1, from the umbilical cord of Healthy People or placenta tissue, extract stem cell, through In vitro culture, amplification, then utilize this type of stem cell to prepare bioactive substance; 2, selected high-quality Herb Gynostemmae Pentaphylli fresh goods, utilizes the various active materials such as modern ultralow temperature nanometer abstraction technique high efficiency extraction saponin, flavonoid, polysaccharide, and at utmost keeps its biological activity; 3, screen full ripe Ganoderma spore, by the modern times activation technique of sprouting, make the biotransformation of the various active materials such as its triterpenoid compound, alkaloid, Ganoderma sterol and organized enzyme and be promoted to peak value, its bioactive substance is extracted in the nanotechnology of recycling ultralow temperature; 4, by people's derived stem cell bioactive extracts, Herb Gynostemmae Pentaphylli bioactive extracts and Ganoderma spore bioactive extracts three compatibility formation compositions application by a certain percentage.
Background technology
Country's " 12 " great ad hoc planning points out that regenerative medicine is the core of future medicine revolution, and stem cells technology is the core of regenerative medicine, and deep development stem cell has Great significance.
Stem cell (stem cell) is that a class has self renewal, highly propagation and the cell colony of multi-lineage potential, is " forefathers' cell ", " the omnipotent cell " that forms the various tissues of human body, organ.Embryonic stem cell and adult stem cell can be divided into according to its stage of development, according to differentiation potential size, myeloid-lymphoid stem cell, pluripotent stem cell and unipotent stem cell can be divided into again.
Stem cell can produce and secrete large number of biological active factors, these stem cell biological active factorses can Effective Regulation body cell signal conduction, activating human body stem cell, and then physiological reparation or alternative body injury, pathological changes and old and feeble cell.For example stem cell can produce stem cell factor (SCF), nerve growth factor (NGF), stromal cell source property somatomedin (SDF), vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin like growth factor (IGF), epidermal growth factor (EGF), interleukin 6 and 7 (IL-6 and IL-7), megakaryocyte colony stimulating factor (M-CSF), tumor necrosis factor (TNF), the factors such as interferon (IFN), wherein the most cells factor all has promotion cell proliferation, differentiation, the inflammation-inhibiting factor and anti-apoptosis effect are closely related etc., stem cell can also produce natural immunity albumen, such as IgG, IgA, IgM, IgD, IgE etc., thus resist or repair self and the damage of extraneous factor to body cell.Research shows human bone marrow, blood vessel, fatty tissue, skeletal muscle, heart and brain, and the upper Intradermal that is also included within lung, liver, pancreas, digestive tract, skin, retina, breast, ovary, prostate and testis is all separable to corresponding stem cell.In theory, stem cell can be repaired the tissue defect at any position of health, can utilize stem cell to go to treat genetic diseases, malignant disease and degenerated disease etc.On October 10th, 2007, one piece of research paper of international artery cell authority magazine < < Cell > > series publication < < < < Cell-Stem Cell > > discloses stem cell function reduction and can cause mankind aging, and proves that stem cell aging process is subject to the external and common regulation and control of intrinsic factor.Up-to-date scientific research discloses, and genesis mechanism and the stem cell regenerating reparation of cardiovascular disease, diabetes etc. have substantial connection.
Therefore, utilize the bioactie agent of stem cell secretion generation or the bioactive substance activating machine soma cell in stem cell, and then by self stem cell physiological reparation or substitute body injury, pathological changes and old and feeble cell, in diseases prevention and treatment and health and beauty field, have broad application prospects.
Herb Gynostemmae Pentaphylli [Gnostemma pentaphyllum (Thunb.) Makino] is Cucurbitaceae Gynostemma Gnostemma plant, has another name called gynostemma pentaphyllum makino, Radix Rhodiolae etc., and there be 11 kind of 2 mutation in this platymiscium whole world Yue13Zhong， China, and wherein 7 Zhong Wei China are peculiar.Within 1986, State Scientific and Technological Commission classifies Herb Gynostemmae Pentaphylli as the Chinese crude drug exploration project first place of Spark Program; calendar year 2001, the State Administration of Forestry was adjusted into National Wild Conservative Plants by Herb Gynostemmae Pentaphylli in < < national key protected wild plants register > >; within 2002, health ministry is formally classified it as new resources medicine-food two-purpose, and is listed in health product list.China's Herb Gynostemmae Pentaphylli aboundresources, DEVELOPMENT PROSPECT is wide.
Herb Gynostemmae Pentaphylli contains the Multiple components such as gypenoside, flavonoid, polysaccharide, aminoacid, trace element.Among the people in order to treat the diseases such as cough, phlegm dyspnea, chronic bronchitis, infectious hepatitis.Modern pharmacology experimental results show that, Herb Gynostemmae Pentaphylli has antitumor, extends cell survival, resisting fatigue, enhancing human body immunity function, antiulcer, control glucocorticoid side effect, regulating lipid metabolism, reduction lipid peroxide, tranquilizing soporific, analgesia, prevents and treats many-sided effects such as senile poliosis, bromhidrosis, U.S. profit skin.Now be developed for blood fat reducing, blood sugar lowering, balancing blood pressure, anticancer, improve immunity and anti-ageing waiting for a long time.
Ganoderma is a kind of medicinal fungi, is the valuable ingredient of Chinese medicine simply in Chinese medicine treasure-house.Ganoderma is widely used in preventing various diseases, has the effect that suppresses tumor and enhancing immunity, and has no side effect, and Ganoderma can effectively prevent oxidative damage and the disease causing thus simultaneously.
Ganoderma spore, as Ganoderma sporophore, also contains a large amount of bioactive substances, and the bioactive substance of Ganoderma spore is higher than Ganoderma sporophore.
In view of upper, we extract bioactive substance at research and development from people's derived stem cell, Herb Gynostemmae Pentaphylli and Ganoderma spore, avoid the loss of high-temperature process to temperature sensitive active substance in extraction process simultaneously, utilize this three classes bioactive substance compositions performance comprehensive effect, activating human body self stem cell comes physiological reparation or alternative body injury, pathological changes and old and feeble histiocyte, regulate blood fat, blood glucose and blood pressure, enhancing human body immunity power, antitumor and slow down aging etc.
Summary of the invention
The object of the invention is: the preparation method that 1, stem cell, Herb Gynostemmae Pentaphylli and Ganoderma spore bioactive substance are provided; 2, preparation method and the application thereof of a kind of stem cell biological active substance, Herb Gynostemmae Pentaphylli bioactive substance and Ganoderma spore bioactive substance compositions are provided.
Technical scheme of the present invention is as follows:
From the umbilical cord of Healthy People or Placenta Hominis, extract stem cell, make it to cultivate in vitro, increase, and maintain high proliferation ability and the stable character of stem cell.Umbilical cord or placenta stem-cell be external was passaged at most for the 5th generation, and stem cell now not only output is large, and character is stable, collects now cell.Adjust cell to suitable concn and be sealed in cryopreservation tube, through liquid nitrogen multigelation, to cell breakage, then by modern biotechnology means such as cryogenic separation, purification, can obtain highly purified stem cell biological active substance group.The whole range request of crossing is without pollutions such as antibacterial, fungus and mycoplasmas.
Selected high-quality Herb Gynostemmae Pentaphylli fresh goods, cleans, goes the pretreatment such as Lao Ye, chopping, and lyophilization, is transferred to certain hour in ultra-low temperature surroundings by dehydration Herb Gynostemmae Pentaphylli, and its sclerosis is become fragile.Take out and carry out respectively ultralow temperature micron order and nanoscale pulverizing, grind, obtain uniform nano-scale particle, and low temperature has guaranteed the lowest loss of active substance.Herb Gynostemmae Pentaphylli nano powder is put in 37 ℃ of pure water of 5 times of weight to ultrasound wave extracting 2 hours, more centrifugal, sucking filtration is separated.Mixed solution is dry through condensation, anhydrates minute, can obtain high activity Herb Gynostemmae Pentaphylli extract.
Utilize seed screening machine to screen Ganoderma spore fresh, ripe, full, homogeneous, being positioned over special-purpose Ganoderma spore sprouts in incubator, under light control bar part, through constant warm water, spray, adopt digital function infrared humidometer constantly to measure the water content of Ganoderma spore, control its water content within the specific limits, utilize determination of oxygen content instrument simultaneously, the ambient oxygen content while sprouting activation by connecting oxygenous steel cylinder control Ganoderma spore.Through the cultivation of sprouting of certain phase, excite the biotransformation of the various active materials such as ganoderma lucidum triterpene compounds, alkaloid, Ganoderma sterol and organized enzyme in spore and be promoted to peak value, biological activity significantly promotes and reaches peak.
The Ganoderma spore that screening is sprouted after activating is transferred to certain hour in ultra-low temperature surroundings immediately, and its sclerosis is become fragile.Ultralow temperature micron order grinds, and shells.Ganoderma spore powder after shelling is placed in nano-level grinder, carries out ultralow temperature fine grinding, obtains uniform nano-scale particle, and low temperature has guaranteed the lowest loss of active substance.Finally, dry through condensation, remove certain moisture, can obtain high-quality Ganoderma spore powder.
Finally, by the bioactive substance extracting from people's derived stem cell, Herb Gynostemmae Pentaphylli and Ganoderma spore compatibility formation according to a certain percentage compositions, said composition is rich in the various active materials such as stem cell factor group, polypeptide, natural immunity albumen, nucleic acid, saponin, flavonoid, polysaccharide, aminoacid, body stem cells hyperplasia, differentiation be can promote, body immunity and anti-aging effects improved.This active compound composition can be made into tablet, powder or capsule, in medical science and healthcare field, has splendid application prospect.
The present invention carries out compatibility by people's derived stem cell, Herb Gynostemmae Pentaphylli and Ganoderma spore bioactive extracts first, synergism on the basis of displaying one's respective advantages, make it the comprehensive effect that performance is larger, thereby regulate better blood fat, blood glucose, blood pressure, improve body immunity, antitumor and anti-ageing waiting for a long time, its effect is better than using separately Stem Cell Activity thing, Herb Gynostemmae Pentaphylli extract or Ganoderma spore powder.
The specific embodiment
The preparation of embodiment 1, umbilical cord or placenta stem-cell bioactive substance
From umbilical cord or the placenta tissue of Healthy People, extract stem cell, this stem cell is mainly mescenchymal stem cell, and the stem cell of acquisition carries out In vitro culture, amplification.Primary cell collect after by the cultivation of going down to posterity of 1: 2 ratio, when cell approximately 80% converges, utilize pancreatin to digest, collect, by identical ratio, go down to posterity, go down to posterity at most and be cultured to for the 5th generation, collected for 5 generations with the cell of interior (comprising for the 5th generation), the stem cell biology character of now collecting is stable, competence for added value is strong, collects the useless culture fluid of changing in cultivation, amplification procedure simultaneously and preserves in-80 ℃ of refrigerators.Detailed process is as follows:
1, gather the umbilicus band of Healthy People;
2, remove blood vessel and outer membrane in umbilical cord;
3, umbilical cord tissue is cut into the fritter of 1～2mm3 size;
4, fritter after shearing is planted in the culture dish that is added with serum-free mescenchymal stem cell culture fluid (diameter is 100mm), be placed in 37 ℃, saturated humidity, 5%CO
2in incubator, cultivate;
5, every 3～4 days half amounts are changed liquid, cultivate 14～16 days Growth of Cells approximately 80% and converge, and collect the useless culture fluid of changing and preserve in-80 ℃ of refrigerators;
6, when cell approximately 80% converges, utilize 0.25% pancreatin to digest cell, with 1～3 * 10
5cells/ml is inoculated in the cultivation of going down to posterity in culture dish, and collects the useless culture fluid of changing and preserve in-80 ℃ of refrigerators;
7, when cell approximately 80% converges, utilize 0.25% pancreatin to digest cell, collect 1st generation cell, by 1: 2 ratio, go down to posterity, be placed in 37 ℃, saturated humidity, 5%CO
2in incubator, cultivate, and collect the useless culture fluid of changing and preserve in-80 ℃ of refrigerators;
If 8 need repeatedly go down to posterity, repeating step 7.;
9, separated, purification obtains high-purity mescenchymal stem cell, utilizes its mescenchymal stem cell purity of FACS Calibur flow cytometry analysis.
10, the umbilical cord or the placenta tissue stem cell concentration that In vitro culture are obtained are adjusted into 5 * 10
6～1 * 10
7individual/milliliter is sealed in cryopreservation tube, through ultra-low temperature liquid nitrogen multigelation 4～5 times, to cell breakage, then by modern biotechnology means such as cryogenic separation, purification, just can obtain bioactive substance group in highly purified stem cell.
The preparation of embodiment 2, Herb Gynostemmae Pentaphylli bioactive substance
1, selected high-quality Herb Gynostemmae Pentaphylli, cleans, goes the pretreatment such as Lao Ye, machinery chopping;
2, pretreated Herb Gynostemmae Pentaphylli is put in freezer dryer and carry out lyophilization, dewater;
3, the Herb Gynostemmae Pentaphylli of lyophilizing is transferred to immediately in ultra-low temperature surroundings (75 ℃～-85 ℃) more than 10 hours, its sclerosis is become fragile;
4, the Herb Gynostemmae Pentaphylli after embrittlement first connects grinding flour by ultralow temperature (60 ℃～-85 ℃) micron order and nanometer;
5, Herb Gynostemmae Pentaphylli nano powder adds 5 times of weight pure water, in 37 ℃, carries out ultrasound wave (30Hz～50Hz) extracting 2 hours;
6, the solution after ultrasonic Treatment is carried out centrifugal, sucking filtration decontamination;
7, supernatant mixed solution is carried out to condensation at low temperature and be dried, anhydrate, obtain Herb Gynostemmae Pentaphylli high activity extract.
The preparation of embodiment 3, Ganoderma spore bioactive substance
1, utilize seed screening machine to screen Ganoderma spore fresh, ripe, full, homogeneous, be positioned over special-purpose Ganoderma spore and sprout in incubator;
2, under light control bar part, through constant warm water (20 ℃～24 ℃), spray the Ganoderma spore in incubator, adopt digital function infrared humidometer constantly to measure the water content of Ganoderma spore, control its water content in 50%～60% scope;
3, utilize determination of oxygen content instrument, measure the oxygen content of Ganoderma spore culture environment, the ambient oxygen content while sprouting activation by connecting oxygenous steel cylinder control Ganoderma spore is in 21%～23% scope;
4, through the cultivation of sprouting of 8～10 hours, according to Ganoderma spore oxygen consumption, determine to excite the biotransformation of the various active materials such as ganoderma lucidum triterpene compounds, alkaloid, Ganoderma sterol and organized enzyme in spore and to be promoted to peak value, biological activity significantly promotes and reaches peak.
5, the Ganoderma spore of screening after sprouting and activating, is transferred in ultra-low temperature surroundings (75 ℃～-85 ℃) more than 10 hours immediately, and its sclerosis is become fragile;
6, ultralow temperature micron order grinds, and shells;
7, the Ganoderma spore powder after shelling is placed in nano-level grinder, carries out ultralow temperature fine grinding, obtains uniform nano-scale particle;
8, nanoscale Ganoderma spore powder is dry through condensation, removes certain moisture, can obtain high-quality Ganoderma spore powder;
Preparation and the application thereof of embodiment 4, stem cell, Herb Gynostemmae Pentaphylli and Ganoderma spore bioactive composition
By from stem cell, Herb Gynostemmae Pentaphylli with the bioactive substance extracting Ganoderma spore according to 1: (5～10): the mass ratio of (10～15) evenly mixes, can obtain desired composition, said composition can be made tablet, powder or capsule and be applied to regulate blood fat, blood glucose and blood pressure, improve body immunity, antitumor and the anti-ageing medical health care field of waiting for a long time.
Claims (10)
1. people's derived stem cell and Herb Gynostemmae Pentaphylli bioactive substance composition and method of making the same, it is characterized in that stem cell biological active substance derives from the stem cell of people's umbilical cord or placenta tissue, is rich in the materials such as stem cell factor group, active polypeptide, immune protein and nucleic acid in it; Herb Gynostemmae Pentaphylli bioactive substance utilizes ultralow temperature nanotechnology extraction, is rich in the various active materials such as saponin, flavonoid, polysaccharide in it; Ganoderma spore bioactive substance derives from the Ganoderma spore of sprouting after activating, and is rich in the various active materials such as triterpenoid compound, alkaloid, Ganoderma sterol and organized enzyme in it.Its preparation method behaviour derived stem cell: extract people's derived stem cell → In vitro culture, amplification → ultra-low temperature liquid nitrogen multigelation to cell breakage → cryogenic separation, purification stem cell biological active substance; Herb Gynostemmae Pentaphylli: selected Herb Gynostemmae Pentaphylli fresh goods → cryotherapy embrittlement → micron and nanoscale powder process → ultrasound wave extracting → centrifugal, be dried → acquisition of sucking filtration → condensation Herb Gynostemmae Pentaphylli extract; Ganoderma spore: shell → nanoscale of the glossy ganoderma spore activating → cryotherapy of sprouting embrittlement → superfine grinding grinding flour.Finally, by three's compatibility formation by a certain percentage compositions, said composition can be applicable to medical health care field.
2. stem cell according to claim 1, is characterized in that deriving from umbilical cord or the Placenta Hominis of Healthy People, through In vitro culture, amplification, obtains.
3. stem cell according to claim 2, is characterized in that including but not limited to mescenchymal stem cell.
4. stem cell biological active substance according to claim 1, is characterized in that comprising cellular content and excreted factor.
5. Herb Gynostemmae Pentaphylli according to claim 1, is characterized in that through cryotherapy embrittlement.
6. Herb Gynostemmae Pentaphylli bioactive substance according to claim 1, it is characterized in that nano-milled, ultrasound wave extracting, condensation dry extraction, is rich in high-activity biological material.
7. Ganoderma spore according to claim 1, is characterized in that through the activation of sprouting.
8. Ganoderma spore bioactive substance according to claim 1, is characterized in that extracting in the Ganoderma spore from sprouting activating, and is rich in the high-activity biological factor.
9. compositions according to claim 1, it is characterized in that people's derived stem cell, Herb Gynostemmae Pentaphylli with Ganoderma spore bioactive substance by 1: (5～10): the mass ratio of (10～15) evenly mixes, but be not limited to this ratio.
10. medical health care purposes according to claim 1, its feature is mainly to regulate blood fat, blood glucose and blood pressure, improves immunity of organisms, antitumor and anti-ageing waiting for a long time.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105902598A (en) * | 2016-05-31 | 2016-08-31 | 安徽惠恩生物科技股份有限公司 | Human-derived stem cell and gynostemma pentaphyllum bioactive substance composition and preparation method thereof |
| CN113730439A (en) * | 2021-09-09 | 2021-12-03 | 陕西中鸿瑞康健康管理有限公司 | Stem cell factor freeze-dried powder capable of reducing triglyceride and preparation method and application thereof |
-
2013
- 2013-10-13 CN CN201310491336.8A patent/CN103565901A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105902598A (en) * | 2016-05-31 | 2016-08-31 | 安徽惠恩生物科技股份有限公司 | Human-derived stem cell and gynostemma pentaphyllum bioactive substance composition and preparation method thereof |
| CN113730439A (en) * | 2021-09-09 | 2021-12-03 | 陕西中鸿瑞康健康管理有限公司 | Stem cell factor freeze-dried powder capable of reducing triglyceride and preparation method and application thereof |
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Application publication date: 20140212 |