CN1994338A - Novel use of Sarcandra glabra aqueous extract - Google Patents
Novel use of Sarcandra glabra aqueous extract Download PDFInfo
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- CN1994338A CN1994338A CNA2006101242253A CN200610124225A CN1994338A CN 1994338 A CN1994338 A CN 1994338A CN A2006101242253 A CNA2006101242253 A CN A2006101242253A CN 200610124225 A CN200610124225 A CN 200610124225A CN 1994338 A CN1994338 A CN 1994338A
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Abstract
The invention relates to an application of sarcandra glabra water extractive, which can prepare the drug that treats immunity reduction caused by load change, health-care food, food additive, etc, wherein said extractive can adjust the immunity organ reduction, improve spleen cells; improve activity of NK cell; and release the ration disorder of T cell group of lymphocyte. The inventive extractive has high safety, while it can be added 0.001-100% into variable food additive, etc.
Description
Technical field
The present invention relates to field of medicaments, specifically is the application of natural plants Herba Pileae Scriptae extract.
Background technology
Herba Pileae Scriptae is meant Chloranthaceae plant plait Corallium Japonicum Kishinouye Sarcandra glabra (Thunb.) Nakai, claim Herba Pileae Scriptae, Herba Pileae Scriptae, Herba Sarcandrae, synthetism tea etc. again, be perennial evergreen draft or undershrub, grow in the dark and damp place of jungle, be distributed in East China, Central-South, southwestern, aboundresources, hyoscine among the people, with a long history
The Pharmacopoeia of the People's Republic of China (version (an one) in 2005) records it and uses as legal medical material.Clinically, Herba Pileae Scriptae has curative effect (http://www.usetm.com/y12/) preferably to multiple active chronic inflammations such as tonsillitis, otitis media, laryngitis, pneumonia, gastroenteritis, appendicitis, paronychia, pustule.In addition, Herba Pileae Scriptae also shows certain effect to cancer of pancreas, nasopharyngeal carcinoma, bladder cancer, thyroid carcinoma, gastric cancer, rectal cancer and the esophageal carcinoma, also is used for the treatment of diseases such as rheumatic arthralgia, rheumatoid arthritis (new medical science 19778 (7): 320).
Pharmacological research in recent years proves that Herba Pileae Scriptae has broad-spectrum antibacterial action, wherein to staphylococcus aureus, dysentery bacterium I type for extremely quick, to golden Portugal persister, Bao Shi dysentery bacterium C-2 type, Bacillus typhi H
901, the paratyphoid fever type is Gao Min, to bacterium flexneri III type, escherichia coli, bacillus pyocyaneus, also (the Chinese herbal medicine communication 1972 (6): 61) of certain inhibitory action arranged.Some studies have shown that Herba Pileae Scriptae has significant inhibition to make (Colleges Of Traditional Chinese Medicine Of Guangxi's journal 200017 (1): 50-52) to the mice inflammation due to Oleum Tiglii and the intersection dish glue.
The Herba Pileae Scriptae product of having developed at present has: QINGRE XIAOYANNING, FUFANG CAOSHANHU HANPIAN, Herba Pileae Scriptae toothpaste, ZHONGJIEFENG PIAN, Sarcandra glaber chewing gum, Herba Pileae Scriptae collutory, Herba Pileae Scriptae teabag etc.Wherein " QINGRE XIAOYANNING " is simple prescription preparation, is that Herba Pileae Scriptae is made with water extract, is used for influenza clinically, pharyngolaryngitis, pneumonia, bacillary dysentery, acute gastroenteritis, appendicitis, burn, the skin infection abscess, cellulitis, the favor of being cured the patient for many years deeply occupies market widely.
Because the Herba Pileae Scriptae water extract has safety preferably, the true border of The acute toxicity tests is nontoxic, mouse sperm deformity test, bone marrow cell micronucleus are tested, Salmonella reversion test is all negative, find no the effect (Guiyang Medical College journal, 199823 (1): 43-44) that cause mammalian somatic cell and sexual cell distortion and bacterial gene sudden change.Thereby think Herba Pileae Scriptae as edible, and safer aspect mutagenic action, therefore be necessary that the new purposes Herba Pileae Scriptae of developing water extract does not have acute toxicity.
Along with the progress of science and technology and the quickening of rhythm of life, people need bear all pressure from different aspect in order to adapt to the intensive day by day competition of society, make people usually be in stress state, by stress and the problem that causes is also following.
Anti-stress medicine can weaken stress be to the effect of body, reduce body to stress reactivity, usually there are neuroleptanalgesiss agent (chlorpromazine, Lacumin, trifluoperazine, haloperidol), tranquilizer (chlorine dihydro aminomethyl phenyl benzodiazepine ketone, phenazepam) and these medicine majorities of tranquilizer (Benactyzire, sodium bromide, dibasole) to have side effect and on clinical using dosage, are subjected to certain limitation, on effect, also are confined to temporary alleviation more, and can not fundamentally reach therapeutic purposes.We develop new anti-stress medicine for this reason.
Summary of the invention
The object of the present invention is to provide the Herba Pileae Scriptae water extract is raw material, is used to prepare treatment or improves medicine, the health food of the immunocompromised of the cause of stress loading, the application of food additive.
The Herba Pileae Scriptae water extract is to adopt the technology of pharmaceuticals industry routine to make, and during extraction, the ratio of Herba Pileae Scriptae and aqueous solvent does not have specific limited, but generally Herba Pileae Scriptae and water weight ratio be advisable with 1: 5～30 scopes.The present invention does not have specific limited to extracting temperature, and is all applicable to the solvent boiling point scope from room temperature.Extraction time, scope could from 10 seconds to 24 hours.
The use amount of Herba Pileae Scriptae water extract provided by the invention accounts for 0.001～100% weight ratio generally in dry product weight in the various finished products of medicine or health food or food additive.To add concentration generally comparatively suitable in 0.01% to 50% (W/W) scope when preparation beverage time institute.In this concentration range, Herba Pileae Scriptae extract is suitable for directly taking, and is accepted by consumer easily.Work as the increase of extract when Herba Pileae Scriptae, pained sensation may occur along with addition.Its solution can be changed dosage form according to the requirement of market demand and consumer, as using dosage forms such as tablet, granule, capsule.Low concentration Herba Pileae Scriptae extract solution can be used as daily beverage and offers consumer.
The Herba Pileae Scriptae water extract is made preparation after also can adding pharmaceutical carriers such as excipient commonly used.Also can add preparation additives such as some antiseptic, antioxidant (antioxidant), coloring agent, sweeting agent in case of necessity.Relatively Shi Yi excipient can be considered with lactose, white sugar, D-mannitol, starch, crystalline cellulose etc.Lubricant can be used magnesium stearate, calcium stearate, Pulvis Talci etc.Solvent can be used Purified Water, ethanol, propylene glycol etc.Antiseptic available chlorine butanols, benzylalcohol, dihydroxy acetic acid etc.Antioxidant is more suitable with sulphite, ascorbic acid etc. usually.
The Herba Pileae Scriptae water extract, can get the extract of its extractum shape or Powdered dry product add to be used in the goods oral.The Herba Pileae Scriptae water extract also can with make beverage, as Sarcandra glabra beverage, soda pop, fruit drink, lactobacillus beverage, sports drink, soyabean milk etc.Herba Pileae Scriptae extract food it is also conceivable that daily breads such as making cookies, chocolate-like, confection class, chewing gum class, snack class, fruit jelly snack categories, bread, bean curd, yoghourt.The said Herba Pileae Scriptae water extract of the present invention also can be made skin preparations for extenal use.As dosage forms such as aerosol, liquid preparation, Emulsion, ointment, semifluid preparation, liniment, lotions.
The said Herba Pileae Scriptae water extract of the present invention can multiple peroral dosage form such as medicine, health food, food additive is used for the treatment of and improve the various clinical symptoms of the immunocompromised that causes of stress loading.
Animal test results shows that the stimulation that directly acts on body can cause the inhibition of immune function.Use the strong inflexible property stress test that rat or mice carried out to show, thymus and spleen atrophy appear behind strong inflexible 24h, the cell number that is combined into rosettes with sheep red blood cell (SRBC) reduces, NK cytoactive and phagocyte active low (animal medicine progress, 1,999 20 (3): 32-34).Stress cause that the index that immune system changes mainly contains two classes: the one, the scalar type index refers to the variation of white blood cell count, mainly is the variation of natural killer cell, lymph T cell, B cell quantity.The 2nd, the functional type index comprises cellular immune function index and humoral immune function index.The former refers to the variation of activity of immune cells and multiplication capacity, and the latter refers to the generation and the variation of the product (as antibody) of immunocyte.Main cellular immune function index is (the http://www.psychcn.com/) such as multiplication capacities of NK cell activity, T cell, B cell.Wherein, the variation of the quality and quantity of the main NK of showing as cell and T cell due to the immunosuppressant that stress cause.Therefore, the active matter of research improvement and raising NK cell and function is of value to the body anti-stress effect undoubtedly.
To stress load body occurred is fatigue reaction, its topmost form of expression is chronic fatigue syndrome (chronic fatigue syndrome CFS).Chronic fatigue syndrome be a kind of be the systemic disease of main performance with the chronic fatigue, be to show as main syndrome so that fatigue, low grade fever, difficult concentrating, hypomnesis, sleep disorder and depression etc. are non-specific.There is the disorder of function of immune system in chronic fatigue syndrome patient.Therefore, there is the people also chronic fatigue syndrome to be called " confirmed fatigue immune dysfunction syndrome ".
It is low that chronic fatigue syndrome mainly shows as cellular immune function, NK cell number or active reduction, this prompting chronic fatigue syndrome patient's nonspecific immunity hypofunction.In addition, chronic fatigue syndrome also exists T cell function, lymphopoiesis function, mononuclear cell to engulf the change of antigenic capacity decline and T cell subsets number and ratio.The decline of above-mentioned cellular immune function can cause the removing ability of body and reply the ability drop of exotic antigen, thereby makes virus can hide in the body for a long time or exotic antigen can not be eliminated.T cell subsets number and ratio change can cause that also cytokine changes, and then causes the change of cellular immunization and humoral immune function.
Infection (anti-infectious immunity) is body opposing causal organism and harmful product thereof, keeps the stable function of organismic internal environment.The power of anti-infection ability also depends on body's immunological function.The immunologic cytotoxicity defence line, first road in NK cellularity resisting pathogenic microbes stage.The NK cell not only has antitumor, antiviral and anti-microbial effect, and by discharging some cytokines, plays an important role in inflammation and immunne response process.The vigor that improves the NK cell is one of important indicator of anti-infectives.
According to the pharmacological evaluation of following examples, can estimate Herba Pileae Scriptae anti-stress effect and immunity effect action effect with the animal model of contained stress induction mouse immune underactivity.Can prove oral administration from experiment, the Herba Pileae Scriptae water extract can effectively be regulated the atrophy of immune organ that causes in response to swashing; The spleen cell decreased number; The lymphopoiesis ability drop; The ratio disorder of T cell subsets in the lymphocyte; Strengthen the NK cell killing activity.Therefore the Herba Pileae Scriptae water extract can be used for preparing treatment or improves because the chronic fatigue syndrome that immunocompromised causes and the medicine or health food or the food additive that infect disease; Mainly be medicine or health food or the food additive that is used to prepare treatment or improves the immunocompromised that causes that the NK cytoactive lowly causes of to load.
The Herba Pileae Scriptae water extract is a kind of safe, can pass through the human body immunity improving system, thus reach prevention, improve and treatment stress, fatigue, infection, immunocompromised is used to prepare medicine that can take for a long time, safe without toxic side effect, food additive, health food and constituent thereof.The Herba Pileae Scriptae water extract reaches 125mg/kg when above at dispensing dosage, promptly demonstrates effective improvement in response to swashing the various hypoimmunity symptoms that load causes.The dosage of Herba Pileae Scriptae water extract among the present invention can be regulated according to application person's conditions such as health status, age, sex and body weight.Adult's average weight is as calculating with 60Kg, Herba Pileae Scriptae and Herba Pileae Scriptae extract day oral dose generally can consider in 0.5g to 10g scope part vic.Certainly there are not problems such as side effect more than the 10g yet.
The present invention can prove the new purposes of Sarcandra glabra aqueous extract by following examples, but is not limited only to following examples.
The specific embodiment
The preparation of embodiment 1 Herba Pileae Scriptae extract
Exsiccant Herba Pileae Scriptae medical material 100g uses pulverizer to pulverize, and adds water 1200ml and boils 60 minutes at 100 ℃, filter, get extracting solution, filtering residue adds 800ml water, boils 30 minutes at 100 ℃, filter, get extracting solution, the extracting solution that merges secondary after gauze filters, through 1000rpm centrifugal 5 minutes, get supernatant concentration after the separation, lyophilization becomes dry extract promptly to get Sarcandra glabra aqueous extract 20g.This Herba Pileae Scriptae water is carried product and is used for embodiments of the invention 2～5.
Embodiment 2 Herba Pileae Scriptae water extract promote to restrain stress cause the low restitution of mouse immune organ index
Laboratory animal: buy male C57BL/6 mice in 6 ages in week, 23 ± 2 ℃ of raising temperatures, 800Lux intensity of illumination illumination 12h/d (7:00-19:00) is used for experiment after raising a week.
Test specimen: embodiment 1 gained Sarcandra glabra aqueous extract dry extract 500mg is dissolved in distilled water 10ml
Stress model: restrain the stress group mice and be fixed on that restrain in the 50mL conical tube that is equipped with air-vent stress 12h, following examples all are with this identical Stress model
Test method: mice is divided 4 groups at random, and 5 every group, be respectively matched group, restrain the Stress model group, the metering group is hanged down in administration: per os gastric infusion 125mgkg once a day
-1The Herba Pileae Scriptae water extract, the high metering group of administration: gastric infusion 500mgkg once a day
-1The Herba Pileae Scriptae water extract.Except that matched group, all the other each group restrain stress, and from restraining the previous day, model group and matched group are given with the capacity aqueous solution, successive administration is after 5 days, mice taken off vertebra is put to death and the taking-up spleen is weighed.
Result of the test (table 1):
| Group | Index and spleen index (%) |
| Normal control model contrast 125mg/kg 500mg/kg | 0.37±0.03 0.3±0.02 0.41±0.03* 0.44±0.04** |
Annotate: * represents p＜0.05, and * * represents p＜0.01 and stress the significant difference conclusion be arranged the matched group ratio: oral administration 125mgkg
-1And 500mgkg
-1Herba Pileae Scriptae extract demonstrates to alleviate to be restrained the index and spleen index that stress cause and reduces, and proving that the Herba Pileae Scriptae water extract can improve stress be to the damaging action of immune organ.
Embodiment 3 Herba Pileae Scriptae water extract promote to restrain stress cause the low restitution of mouse spleen cell
Experimental animal: 5 ages in week male C57BL/6 mice, 23 ± 2 ℃ of raising temperatures, 800Lux intensity of illumination illumination 12h/d (7:00-19:00) is used for experiment after raising a week.
Test specimen: embodiment 1 gained Sarcandra glabra aqueous extract dry extract 500mg is dissolved in distilled water 10ml
Test method: mice is divided into matched group at random, restrains the Stress model group, 125mgkg
-1And 500mgkg
-1Herba Pileae Scriptae extract administration group, every group of 5 mices are divided into 4 groups.Except that matched group, all the other each group is restrained stress 12h, and from restraining the previous day, respectively give with capacity sterilized water once a day by gastric infusion, model group and matched group for experimental group, after continuous 4 days, mice is taken off vertebra put to death and take out spleen, in PBS solution, separate unicellular with tissue grinder, filter PBS standardize solution cell counting behind the 10mL through 200 order copper mesh.
Result of the test (table 2):
| Group | The total cell number (* 10 of spleen 7) |
| Normal control model contrast 125mg/kg 500mg/kg | 11.7±0.7 10.3±0.3 12.5±0.5* 12.2±0.2* |
Annotate: * represents p＜0.05 and stress significant difference be arranged the matched group ratio
Conclusion: oral administration 125mgkg
-1And 500mgkg
-1Herba Pileae Scriptae extract demonstrates to alleviate to a certain extent restrains (mice) spleen cell number minimizing that stress cause.Proof Herba Pileae Scriptae water extract can improve restrain stress cause spleen cell lowly act on, illustrate that Herba Pileae Scriptae has the effect of the immunocompromised of improvement, easily cause fatigue, various infection disease and infection secondary disease etc. thereby improve immunocompromised.
Embodiment 4 Herba Pileae Scriptae water extract promote to restrain stress cause the low restitution of mouse spleen T lymphopoiesis ability
Laboratory animal: 5 ages in week male C57BL/6 mice, 23 ± 2 ℃ of raising temperatures, the 800Lux 12h/d (7:00-19:00) that throw light on is used for testing after raising a week.
Test specimen: embodiment 1 gained Sarcandra glabra aqueous extract dry extract 500mg is dissolved in distilled water 10ml
Test method: mice is divided into matched group at random, restrains the Stress model group, 125mgkg
-1And 500mgkg
-1Herba Pileae Scriptae extract administration group, every group of 5 mices are divided into 4 groups.Except that matched group, all the other each group is restrained stress 12h, and from restraining the previous day, respectively give with capacity aqueous solution once a day by gastric infusion, model group and matched group for experimental group, after continuous 4 days, mice is taken off vertebra put to death and take out spleen, in PBS solution, separate unicellular with tissue grinder, filter 1500rpmmin through 200 order copper mesh
-1Centrifugal 5min abandons supernatant.Add low the opening of 3mL Tris-HCl (pH7.65) buffer solution and handle an erythrocyte, leave standstill and add PBS liquid 5mL behind the 2min and stop the erythrocyte splitting reaction.1500rpmmin
-1Abandon supernatant behind the centrifugal 5min, change clothes twice with PBS liquid after, add the 10%FCS-RPMI1640 culture fluid and adjust cell concentration to 5 * 10
6CellsmL
-1Each sample lymphocyte suspension (5 * 10
6CellsmL
-1) get 100 μ L respectively and join in the flow cytometry pipe and carry out, add the anti-CD3-FITC antibody labeling of 5 μ L.The room temperature black out adds 500 μ L PBS after hatching 20min, vibration mixing 1500rpmmin
-1Abandon supernatant behind the centrifugal 5min, the back is used to detect per 5 * 10 with 500 μ L PBS vibration mixing
6CD3 in the individual cell
+The cell percentage composition calculates the T lymphocyte (CD3 of whole spleen
+) number.
Result of the test (table 3):
| Group | Splenic T lymphocyte number (* 10 7) |
| Normal control model contrast 125mg/kg 500mg/kg | 3.30±0.21 3.01±3.2 3.80±2.6* 3.84±2.8* |
Annotate: * represents p＜0.05 and stress significant difference be arranged the matched group ratio in the table
Conclusion: oral administration 125mgkg
-1And 500mgkg
-1Herba Pileae Scriptae extract shows and can alleviate the contained splenic T lymphocyte number minimizing that stress cause, proof Herba Pileae Scriptae water extract can improve restrain stress cause immunosuppressive action, improve the fatigue of immunosuppressant cause, various infection disease and infect secondary disease etc.
Embodiment 5 Herba Pileae Scriptae water extract are to restraining the restitution of mouse spleen t lymphocyte subset group ratio disorder that stress cause
Laboratory animal: 5 ages in week male C57BL/6 mice, 23 ± 2 ℃ of raising temperatures, the 800Lux 12h/d (7:00-19:00) that throw light on is used for testing after raising a week.
Laboratory sample: embodiment 1 gained Sarcandra glabra aqueous extract dry extract 500mg is dissolved in distilled water 10ml
Test method: mice is divided into matched group at random, restrains the Stress model group, 125mgkg
-1And 500mgkg
-1Herba Pileae Scriptae extract administration group, every group of 5 mices are divided into 4 groups.Except that matched group, all the other each group is restrained stress 12h, and from restraining the previous day, respectively give with capacity aqueous solution once a day by gastric infusion, model group and matched group for experimental group, after continuous 4 days, mice is taken off vertebra put to death and take out spleen, in PBS solution, separate unicellular with tissue grinder, filter 1500rpmmin through 200 order copper mesh
-1Centrifugal 5min abandons supernatant.Add low the opening of 3mL Tris-HCl (pH7.65) buffer solution and handle an erythrocyte, leave standstill and add PBS liquid 5mL behind the 2min and stop the erythrocyte splitting reaction.1500rpmmin
-1Abandon supernatant behind the centrifugal 5min, change clothes twice with PBS liquid after, add the 10%FCS-RPMI1640 culture fluid and adjust cell concentration to 5 * 10
6CellsmL
-1Each sample lymphocyte suspension (5 * 10
6CellsmL
-1) get 100 μ L respectively and join in two flow cytometry pipes and carry out FITC, PE Two Colour Fluorescence antibody labeling.Wherein a pipe adds 5 μ L anti-cd 3 antibodies and 3 μ L CD4 antibody; Second pipe adds 5 μ L anti-cd 3 antibodies and the anti-CD8 antibody of 3 μ L.The room temperature black out adds 500 μ L PBS after hatching 20min, vibration mixing 1500rpmmin
-1Abandon supernatant behind the centrifugal 5min, the back is used to detect per 5 * 10 with 500 μ L PBS vibration mixing
6CD3 in the individual cell
+CD4
+Cell and CD3
+CD8
+The cell percentage, and calculate CD3
+CD4
+Cell % and CD3
+CD8
+Cell % ratio.The T cell is inhomogenous cell colony, according to its surface marker and functional character, it can be divided into different subgroups, CD4 again
+Cell (Th) and CD8
+Two main subgroups of cell (Ts), CD4
+Cell has auxiliary and inducing function, CD8
+Cell has immunosuppressive action and cell toxicant function, they are cooperation mutually both, functional characteristics is separately arranged again, need finish immunne response jointly and just bring into play immunoloregulation function, immune function of human body has been played important regulatory role so keep the stable of t lymphocyte subset group.
Result of the test (table 4):
| Group | CD4/CD8 |
| Normal control model contrast 125mg/kg 500mg/kg | 1.85±0.2 2.28±0.19** 1.80±0.26 1.95±0.14 |
Annotate: * * represents p＜0.01 in the table, stress significant difference be arranged with the matched group ratio
Conclusion: restrain and to cause CD3
+CD4
+Cell proportion increases, CD3
+CD8
+A kind of dysimmunity state that cell proportion descends, oral administration 125mgkg
-1And 500mgkg
-1Herba Pileae Scriptae extract demonstrates to regulate restrains the splenic T lymphocyte subgroup disorder that stress cause.Proof Herba Pileae Scriptae water extract can improve restrain stress cause the immunologic derangement effect, thereby improve because of the fatigue that immunologic derangement causes, various infection disease and infect secondary disease etc.
Embodiment 6 Herba Pileae Scriptae water extract are lived to the tax of mice NKT sexual cell (natural killer:NK) and are acted on
Laboratory animal: 5 ages in week male C57BL/6 mice, 23 ± 2 ℃ of raising temperatures, lighting hours 12h/d (7:00-19:00) raises and is used for experiment after a week.
Sample: embodiment 1 gained Sarcandra glabra aqueous extract dry extract 500mg is dissolved in distilled water 10m1
Method: mice is divided into matched group at random, restrains the Stress model group, 125mgkg
-1And 500mgkg
-1Herba Pileae Scriptae extract administration group, every group of 5 mices are divided into 4 groups.Except that matched group, all the other each group is restrained stress 12h, and from restraining the previous day, respectively give with capacity aqueous solution once a day by gastric infusion, model group and matched group for experimental group, after continuous 4 days, mice is taken off vertebra put to death and take out spleen, in PBS solution, separate unicellular with tissue grinder, filter 1500rpmmin through 200 order copper mesh
-1Centrifugal 5min abandons supernatant.Add low the opening of 3mL Tris-HCl (pH7.65) buffer solution and handle an erythrocyte, leave standstill and add PBS liquid 5mL behind the 2min and stop the erythrocyte splitting reaction.1500rpmmin
-1Abandon supernatant behind the centrifugal 5min, change clothes twice with PBS liquid after, add the 10%FCS-RPMI1640 culture fluid and adjust cell concentration to 2 * 10
7CellsmL
-1When experiment adopted the target cell of DiO labelling to be killed and wounded breakage, dyestuff PI entered the cell marking nucleus, according to imitating target cell size and DiO dyeing difference, can distinguish the target cell group in FL1/FS figure, and then research target cell death condition, calculated the NK killing activity.Concrete experiment is that the 10%FCS-RPMI1640 culture fluid transfers to 1 * 10 with the splenocyte suspension
6, 5 * 10
5, 2.5 * 10
5And 1.25 * 10
5Cells100 μ L
-1After the concentration, with 100 μ Lwell
-1Add to respectively at the bottom of 96 hole circles in the culture plate as NK lethal effect cell.The 10%FCS-RPMI1640 culture fluid is adjusted YAC-1 cell concentration to 10
6ML
-1, add DiO (10 μ L1mL
-1Cells), at 37 ℃, 5%CO
2After hatching 15min under the condition, use the culture fluid washed twice, cross 300 order nylon wires, adjust cell concentration to 10 with above-mentioned culture fluid
4Cells100 μ L
-1As the NK target cell with 100 μ Lwell
-1Add among the effector lymphocyte (imitate: the target cell ratio was respectively 100: 1, and 50: 1,25: 1 and 12.5: 1).YAC-1 target cell natural mortality rate is 10
4Cells100 μ L
-1Adding 100 μ L culture fluid in the target cell organizes in contrast.37 ℃, 5%CO
2Cultivate after 3 hours, every hole adds 500mgL
-1PI 8 μ L continue to hatch after one hour and take out, and culture plate are placed stop on ice killing and wounding reaction, and keeping in Dark Place is used for the flow cytometer detection.Each porocyte in the culture plate blown afloat gently and change cell suspension to be measured over to the flow cytometer showed pipe after add 500 μ L PBS, 1000 target cells in each sample are done the check and analysis of NK killing activity with flow cytometer.The NK killing activity is represented the cytotoxicity of YAC-1 target cell with the NK cell, and is calculated by following formula:
The killing activity of NK when the present invention calculates 10%cytolysis: LU
10(cytotoxicity is 10% o'clock needed effector lymphocyte's number) needed spleen cell number is that unit comes quantificational expression.
In addition, the present invention has also calculated 10
6NK cell killing activity (the LU1010 that the cells spleen cell has
6Cells
-1) and NK cell killing activity (the LU10spleen cells that had of the whole cell of whole spleen
-1)
Result of the test (table 5):
| LU10·10 6cells -1 | LU10·spleen cells -1 | |
| Normal control model contrast 125mg/kg 250mg/kg | 0.18 0.19 0.21 0.20 | 21.33 19.86 25.72 24.26 |
Conclusion: from result of the test represent and normal group relatively, restrain and stress make LU10spleen cells
-1Descend, promptly restrain the NK cytoactive is descended.Oral administration 125mgkg
-1And 500mgkg
-1The Herba Pileae Scriptae water extract demonstrates when promoting that splenocyte recovers, and also composes the NK cytoactive of living to a certain extent, proves that the Herba Pileae Scriptae water extract can improve because the NK cytoactive that stress cause descends, and has certain immunity effect effect.The decline of NK cytoactive can cause immunity of organisms decline, and tired and infection easily improves tired, as to strengthen anti-infective and infection secondary disease etc. effect so Herba Pileae Scriptae can be used.
Embodiment 6 Sarcandra glabra aqueous extracts are at the various embodiment of preparation medicine or health food or food additive.
1. Herba Pileae Scriptae oral administration dripping pill: Herba Pileae Scriptae medical material 3125g, decocting boil 2 times, and each 1.5 hours, decocting liquid was considered merging, are concentrated in right amount, and spray drying adds a certain amount of drop pill substrate polyethylene glycol 6000, makes the Herba Pileae Scriptae oral administration dripping pill of 1000 parts of doses.
2. the preparation of Herba Pileae Scriptae Tabellae: get Herba Pileae Scriptae medical material 3125g, decocting boils 3 times, and each 1 hour, decocting liquid was considered merging, is concentrated in right amount, and spray drying adds appropriate amount of auxiliary materials, makes particle drying, is pressed into 10000, sugar coating, promptly.
3. compound Caoshanhu skin and complexion improving water: add Herba Pileae Scriptae water extraction dry product 0.001g, Mentholum 0.08g, Borax 0.5g then successively, be heated to 60 ℃ with distilled water and make their dissolvings, be cooled to 38-45 ℃, and then add 2 of glycerol 0.1ml, plant essences, be cooled to 25-30 ℃, add ethanol again and stir, filter and get.
4. Herba Pileae Scriptae hard sugar: Herba Pileae Scriptae water extract dry product 5g, high quality white Saccharum Sinensis Roxb. 65g, first class glucose 28g adds water and makes 10 of Herba Pileae Scriptae hard sugar in right amount.
5. Herba Pileae Scriptae series candied fruit: Herba Pileae Scriptae water extract dry product 1g, Mentholum 0.08g, Ziziphi Spinosae fruit jam 89g, white sugar 5g and milk powder 4g or Semen arachidis hypogaeae 4g or Semen Sesami 4g make the Herba Pileae Scriptae Ziziphi Spinosae cake of multiple odor type.
6. the preparation of Sarcandra glabra health-care drink: 0.4g Herba Pileae Scriptae water is promoted extractum, and 1.6g is mixed by the Saponin and the 0.1g chlorophyll of natural Chinese medicinal herb plant Herb Gynostemmae Pentaphylli plant extract, adds natural mineral water 100ml, and it is formulated to add an amount of sucrose.
7. the preparation of Herba Pileae Scriptae Chewing Gum: 150g Herba Pileae Scriptae water is promoted extractum, Oleum menthae 10ml or Mentholum 4g, and 1000g, edible glue nurse 800g makes 1000.
Claims (4)
1, the Herba Pileae Scriptae water extract be used to prepare treatment improve the medicine of immunocompromised of the cause of stress loading or health food or, the application of food additive, said Herba Pileae Scriptae water extract accounts for 0.001～100% weight ratio in dry product weight in medicine or health food or the various finished products of food additive.
2, Herba Pileae Scriptae water extract according to claim 1 is used to prepare treatment or improves because the medicine of the chronic fatigue syndrome that immunocompromised causes or the application of health food or food additive.
3, be used to prepare treatment or improve the medicine of the infection disease that causes owing to immunocompromised or the application of health food or food additive according to the said Herba Pileae Scriptae water extract of claim 1.
4, be used to prepare treatment according to the described Herba Pileae Scriptae water extract of claim 1～3 or improve the medicine of the immunocompromised that causes that the NK cytoactive lowly causes of stress loading or the application of health food or food additive.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006101242253A CN1994338A (en) | 2006-12-14 | 2006-12-14 | Novel use of Sarcandra glabra aqueous extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006101242253A CN1994338A (en) | 2006-12-14 | 2006-12-14 | Novel use of Sarcandra glabra aqueous extract |
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| Publication Number | Publication Date |
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| CN1994338A true CN1994338A (en) | 2007-07-11 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNA2006101242253A Pending CN1994338A (en) | 2006-12-14 | 2006-12-14 | Novel use of Sarcandra glabra aqueous extract |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526278A (en) * | 2012-03-07 | 2012-07-04 | 贵州宏宇药业有限公司 | Health-care product used for relaxing tendon, activating collaterals and alleviating athletic fatigue and preparation method thereof |
| CN111387386A (en) * | 2020-04-07 | 2020-07-10 | 武汉轻工大学 | Selenium-rich sarcandra glabra enzyme beverage and preparation method and application thereof |
| CN118286278A (en) * | 2024-06-05 | 2024-07-05 | 江西草珊瑚口腔护理用品有限公司 | A method for preparing herba Pileae Scriptae extract by instant pressurizing process |
-
2006
- 2006-12-14 CN CNA2006101242253A patent/CN1994338A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102526278A (en) * | 2012-03-07 | 2012-07-04 | 贵州宏宇药业有限公司 | Health-care product used for relaxing tendon, activating collaterals and alleviating athletic fatigue and preparation method thereof |
| CN102526278B (en) * | 2012-03-07 | 2013-11-20 | 贵州宏宇药业有限公司 | Health-care product used for relaxing tendon, activating collaterals and alleviating athletic fatigue and preparation method thereof |
| CN111387386A (en) * | 2020-04-07 | 2020-07-10 | 武汉轻工大学 | Selenium-rich sarcandra glabra enzyme beverage and preparation method and application thereof |
| CN118286278A (en) * | 2024-06-05 | 2024-07-05 | 江西草珊瑚口腔护理用品有限公司 | A method for preparing herba Pileae Scriptae extract by instant pressurizing process |
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