CN102727872A - Anti-aging and heart and blood vessel protecting soft capsule and preparation method thereof - Google Patents
Anti-aging and heart and blood vessel protecting soft capsule and preparation method thereof Download PDFInfo
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Abstract
The invention relates to an anti-aging and heart and blood vessel protecting soft capsule and a preparation method thereof. The soft capsule comprises a soft capsule body and contents and is characterized in that the contents comprise 40-60% of seabuckthorm seed oil, 1-3% of lecithin, 1-3% of vitamin E, 4-8% of lycopene, 5-9% of propolis, 20-30% of pumpkin seed oil and 6-10% of collagen protein by weight percentage. The contents are evenly mixed, ground and degassed; gelatin, water and glycerol are weighed by the proportion of 100:100:40, heated for melting materials and degassed to obtain a soft capsule body filtrate; the filtrate and the contents are put in a pelleting machine for pelleting by the weight ratio of 1: (2-3) and the finished product is obtained after shaping, drying, pill washing, picking and packaging. By the synergistic effect of the ingredients of the soft capsule, immunity of a human body can be effectively improved. The soft capsule is scientific in dose and has the function of beautifying, resisting aging, protecting heart and blood vessels, tonifying the kidney and controlling nocturnal emission. The preparation method is reasonable in process and liable to industrialized production and has strong operability.
Description
Technical field
The present invention relates to a kind of is the medicine configuration article of characteristic with the capsule preparations, especially a kind of defying age, protection cardiovascular soft capsule and preparation method thereof.
Background technology
The human immune system is the defense mechanism of human body self, and human body is played a protective role.The quickening of Along with people's live and work rhythm, diet is irregular, the too meticulous shortage that causes the special dietary composition of food, contact with chemical substance reason such as increase, cause the human immune system to receive increasing chemistry and physical factor influence.And the human immune system causes damage can cause the generation of fatigue, aging even disease, and particularly the cardiovascular and cerebrovascular vessel sickness rate is increasing year by year in recent years.Therefore, use various medicines with the immunologic function that improves people self and improve with health product and treat the low disease that causes of immune function of human body, for example aging, cardiovascular disease are significant.
We know that Fructrs Hippophae seed oil is the complex of multivitamin and bioactive substance.It can nourishes rough skin, enhance metabolism, antiallergic, and bactericidal antiphlogistic promotes epithelial cell regeneration, and skin is had repair, can keep the sour environment of skin, has stronger permeability, is the important source material of beauty and skin care.Aspect enhancing immunity, Fructrs Hippophae seed oil can obviously strengthen E garland formation rate, serum anti SRBC antibody titer, and the mouse humoral immune cell and the cellular immune function of radiation damage are recovered all to have significant enhancement effect.Through the modern medicine clinical verification, Fructrs Hippophae seed oil has defying age, skin whitening, antiinflammatory granulation promoting, promotes tissue regeneration, antitumor, nourishing the brain and improving intelligence, the protection heart, cerebrovascular, the immune effect of adjusting.
Lecithin is the key component of cell membrane, is the main component of cell self-regeneration.Additional lecithin can improve the regeneration capacity of metabolic capacity, self-healing ability and the antibody tissue of human body, delaying human body caducity.In addition, lecithin can cut grease in emulsifying, and vessel softening and removing peroxide reach unobstructed blood vessel, the effect of blood sugar lowering and blood fat.
Vitamin E is a kind of fat-soluble vitamin, mainly is used as antioxidant, slow down aging.In addition, also can be used as vasodilation and anticoagulant, improve blood circulation.
Utilize Fructrs Hippophae seed oil, lecithin, vitamin E preparation to have the health product of cardiovascular protection effect, have report at present.For example; On July 5th, 2006 disclosed CN 1262284C Chinese invention patent description just disclose have antioxidation, " a kind of Oleum Hippophae health product " of cardiovascular protection effect; Its activated feedstock by following weight portion proportioning is formed: 80～99 parts of purification Fructrs Hippophae seed oil; 0.5～3 part in lecithin, 0.5～3 part of vitamin E.In its preferred version, each activated feedstock weight portion proportioning: 96 parts of purification Fructrs Hippophae seed oil, 1 part in lecithin, vitamin e1 part.In the above-mentioned disclosed file, active constitutive material composition is few, has influenced the effect of antioxidation and cardiovascular protection, and slightly inadequate to the raising aspect of body immunity.
Summary of the invention
In order to overcome prior art in defying age, protection cardiovascular effect unfavorable deficiency, the present invention provide a kind of dosage science, effect good, improve body immunity, have defying age, protection cardiovascular soft capsule of supplementing the kidney to control the nocturnal health care and preparation method thereof.
The technical solution adopted for the present invention to solve the technical problems is: a kind of defying age, protection cardiovascular soft capsule; It is made up of flexible glue utricule and the intravital content of this soft capsule of inclosure, and it is characterized in that: described content is made up of following components in weight percentage: Fructrs Hippophae seed oil 40%～60%, lecithin 1%～3%, vitamin e1 %～3%, lycopene 4%～8%, propolis 5%～9%, Semen Cucurbitae oil 20%～30%, collagen protein 6%～10%; Described flexible glue utricule is to be processed in 100: 100: 40 ratio by gelatin, water, glycerol, and the weight ratio of said flexible glue utricule and said content is: 1: 2～3.
As prioritization scheme, the percentage by weight of the component of described content is respectively: Fructrs Hippophae seed oil 50%, lecithin 2%, vitamin E2 %, lycopene 6%, propolis 7%, Semen Cucurbitae oil 25%, collagen protein 8%.
A kind of defying age, protection cardiovascular preparation of soft capsule method is characterized in that: the following technology of process:
A, batching: according to percentage by weight weighing Fructrs Hippophae seed oil 40%～60%, lecithin 1%～3%, vitamin e1 %～3%, lycopene 4%～8%, propolis 5%～9%, Semen Cucurbitae oil 20%～30%, collagen protein 6%～10%, place material-compound tank, stir;
B, grinding, the degassing: place colloid mill to grind three times the Fructrs Hippophae seed oil that stirs, lecithin, vitamin e1, lycopene, propolis, Semen Cucurbitae oil, the collagen protein of step a gained; Cross 100 mesh sieves; Place vacuum kettle then; Control vacuum-0.06Mpa sloughs gas, gets the content material;
The making of c, flexible glue envelop materials: gelatin, water, glycerol in ratio weighing in 100: 100: 40, in the being shipped to glue jar, are stirred and are warming up to 60～75 ℃; Treat that gelatin all melts, decompression vacuum pumping, control vacuum-0.06Mpa; Slough gas, cross 80 mesh sieves, obtain flexible glue utricule filtrating; Contain in heat-preserving container, subsequent use;
D, pelleting: with the resulting flexible glue utricule of step c filtrating and the resulting content material of step b according to weight ratio: 1: 2～3 ratio is sent in the pellet press, suppresses soft capsule;
E, typing drying: with 22～26 ℃ of the resulting soft capsule control of steps d temperature, humidity is 30～40%, after static dry 24 hours, goes out ball;
F, wash ball: it is the surperficial oil stain of ethanol flush away utricule 90% or more that the resulting soft capsule of step e is adopted concentration;
G, choosing are chosen: the soft capsule of the resulting flush away of step f surface oil stain is selected chooses, remove non-eurymeric ball, the eurymeric soft capsule;
H, packing, finished product: with the bottling of the resulting eurymeric soft capsule of step g, seal, finished product.
Defying age of the present invention, protection cardiovascular soft capsule are to have carried out Fructrs Hippophae seed oil, lecithin and vitamin E with antioxidation, vascular protection effect reasonably combined; Increase the content of vitamin E and lecithin, and further chosen lycopene, propolis, Semen Cucurbitae oil and the collagen protein that has defying age, protects the cardiovascular effect.
In the present invention, its lycopene of selecting for use contains 11 conjugated double bonds and 2 unconjugated double bonds on molecular structure, in carotenoid; It has the strongest antioxidant activity, and its effect of removing free radical outclass other carotenoid and vitamin E, and the speed constant of its cancellation singlet oxygen is 100 times of vitamin E; It is through strengthening the activity of antioxidase; Improve the oxidation resistance of body, keep the cell homergy, and can strengthen the normal cell gap junction communication; Promote the interaction between phagocyte, lymphocyte; Through excretory cell activation factor activating cell, finally show phagocytic activity and LT promotion, raise immunity.Lycopene has above superior physiological function, makes it not only have anticancer, as to press down cancer effect, and various adult diseases such as angiocardiopathy preventing, arteriosclerosis, prevention aging and enhancing human body immunity power etc. are had important function; Its selected propolis contains more than 70 kind of flavone compound, organic acid, mineral, trace element, vitamin and aminoacid.Recent studies proves, propolis contained abundant and unique biological active substance makes it have multiple functions such as antibiotic, antiinflammatory, antipruritic, antioxidation, enhance immunity, blood sugar lowering, blood fat reducing, antitumor; Zinc in its selected Semen Cucurbitae oil, magnesium, calcium, phosphorus content are high; Especially contain multiple bioactive substances such as abundant unsaturated fatty acid, plant sterol, aminoacid, vitamin, mineral; Can blood sugar lowering, cholesterol, diabetes and the heart, cerebrovascular disease are had prevention and health-care effect.In addition, contain a kind of androgen's active bio catalyzer contact agent composition in the Semen Cucurbitae oil, can eliminate prostatic initial stage swelling, urinary system and prostatic hyperplasia are had good curing and preventive effect; Its selected collagen protein is the essential protein of tissue, can effectively replenish needed by human body, supports the collagen peptide bond and the elasticated net of skin, prevents netted structural deterioration of spiral and skin histology oxidation, atrophy, subsides slow down aging.
Key of the present invention is the proportioning with each amounts of components of choosing of the intravital content component of soft capsule.Its lycopene of selecting for use, propolis, Fructrs Hippophae seed oil, Semen Cucurbitae oil, collagen protein, lecithin and vitamin E combined effect can reach best defying age, protection cardiovascular effect.Compared with prior art, its dosage science, defying age, the protection cardiovascular effective, effectively improved body immunity, and had the function of supplementing the kidney to control the nocturnal.Method for preparing provided by the present invention, its technology is reasonable, and is workable, is prone to accomplish scale production.
The specific embodiment
Below in conjunction with embodiment the present invention is further specified.
Embodiment 1
A kind of defying age, protection cardiovascular preparation of soft capsule method, the following technology of process:
A, batching: weighing Fructrs Hippophae seed oil 50kg, lecithin 2kg, vitamin E2 kg, lycopene 6kg, propolis 7kg, Semen Cucurbitae oil 25kg, collagen protein 8kg, place material-compound tank, stir;
B, grinding, the degassing: place colloid mill to grind three times the Fructrs Hippophae seed oil that stirs, lecithin, vitamin e1, lycopene, propolis, Semen Cucurbitae oil, the collagen protein of step a gained; Cross 100 mesh sieves; Place vacuum kettle then; Control vacuum-0.06Mpa sloughs gas, gets the content material;
The making of c, flexible glue envelop materials: in weighing gelatin 16.6kg, water 16.6kg, being shipped to of the glycerol 6.6kg glue jar, stir and be warming up to 75 ℃, treat that gelatin all melts; Decompression vacuum pumping, control vacuum-0.06Mpa sloughs gas; Cross 80 mesh sieves; Obtain flexible glue utricule filtrating, contain in heat-preserving container, subsequent use;
D, pelleting: with the resulting flexible glue utricule of step c filtrating and the resulting content material of step b according to weight ratio: 1: 2.51 ratio is sent in the pellet press, suppresses soft capsule;
E, typing drying: with 26 ℃ of the resulting soft capsule control of steps d temperature, humidity is 30%, after static dry 24 hours, goes out ball;
F, wash ball: it is the oil stain on 92% ethanol flush away utricule surface that the resulting soft capsule of step e is adopted concentration;
G, choosing are chosen: the soft capsule of the resulting flush away of step f surface oil stain is selected chooses, remove non-eurymeric ball, the eurymeric soft capsule;
H, packing, finished product: with the bottling of the resulting eurymeric soft capsule of step g, seal, finished product.
In the present embodiment, the percentage by weight of the intravital content of soft capsule is respectively: Fructrs Hippophae seed oil 50%, lecithin 2%, vitamin E2 %, lycopene 6%, propolis 7%, Semen Cucurbitae oil 25%, collagen protein 8%; The flexible glue utricule is to be processed in 100: 100: 40 ratio by gelatin, water, glycerol.The soft capsule dosage science of gained of the present invention, defying age, protection cardiovascular are effective, and have the supplementing the kidney to control the nocturnal function; Its preparation method technology is reasonable, and is workable, is prone to accomplish scale production.
Embodiment 2
A kind of defying age, protection cardiovascular preparation of soft capsule method, the following technology of process:
A, batching: weighing Fructrs Hippophae seed oil 59kg, lecithin 3kg, vitamin E 3kg, lycopene 4kg, propolis 5kg, Semen Cucurbitae oil 20kg, collagen protein 6kg, place material-compound tank, stir;
B, grinding, the degassing: place colloid mill to grind three times the Fructrs Hippophae seed oil that stirs, lecithin, vitamin e1, lycopene, propolis, Semen Cucurbitae oil, the collagen protein of step a gained; Cross 100 mesh sieves; Place vacuum kettle then; Control vacuum-0.06Mpa sloughs gas, gets the content material;
The making of c, flexible glue envelop materials: in weighing gelatin 13.87kg, water 13.87kg, being shipped to of the glycerol 5.55kg glue jar, stir and be warming up to 65 ℃, treat that gelatin all melts; Decompression vacuum pumping, control vacuum-0.06Mpa sloughs gas; Cross 80 mesh sieves then; Obtain flexible glue utricule filtrating, contain in heat-preserving container, subsequent use;
D, pelleting: with the resulting flexible glue utricule of step c filtrating and the resulting content material of step b according to weight ratio: 1: 3 ratio is sent in the pellet press, suppresses soft capsule;
E, typing drying: with 24 ℃ of the resulting soft capsule control of steps d temperature, humidity is 40%, after static dry 24 hours, goes out ball;
F, wash ball: it is the oil stain on 95% ethanol flush away utricule surface that the resulting soft capsule of step e is adopted concentration;
G, choosing are chosen: the soft capsule of the resulting flush away of step f surface oil stain is selected chooses, remove non-eurymeric ball, the eurymeric soft capsule;
H, packing, finished product: with the bottling of the resulting eurymeric soft capsule of step g, seal, finished product.
In the present embodiment, the percentage by weight of the intravital content of soft capsule is respectively: Fructrs Hippophae seed oil 59%, lecithin 3%, vitamin E 3%, lycopene 4%, propolis 5%, Semen Cucurbitae oil 20%, collagen protein 6%; The flexible glue utricule is to be processed in 100: 100: 40 ratio by gelatin, water, glycerol.
Embodiment 3
A kind of defying age, protection cardiovascular preparation of soft capsule method, the following technology of process:
A, batching: weighing Fructrs Hippophae seed oil 41kg, lecithin 1kg, vitamin e1 kg, lycopene 8kg, propolis 9kg, Semen Cucurbitae oil 30kg, collagen protein 10kg, place material-compound tank, stir;
B, grinding, the degassing: place colloid mill to grind three times the Fructrs Hippophae seed oil that stirs, lecithin, vitamin e1, lycopene, propolis, Semen Cucurbitae oil, the collagen protein of step a gained; Cross 100 mesh sieves; Place vacuum kettle then; Control vacuum-0.06Mpa sloughs gas, gets the content material;
The making of c, flexible glue envelop materials: in weighing gelatin 20.8kg, water 20.8kg, being shipped to of the glycerol 8.3kg glue jar, stir and be warming up to 60 ℃, treat that gelatin all melts; Decompression vacuum pumping, control vacuum-0.06Mpa sloughs gas; Cross 80 mesh sieves then; Obtain flexible glue utricule filtrating, contain in heat-preserving container, subsequent use;
D, pelleting: with the resulting flexible glue utricule of step c filtrating and the resulting content material of step b according to weight ratio: 1: 2 ratio is sent in the pellet press, suppresses soft capsule;
E, typing drying: with 22 ℃ of the resulting soft capsule control of steps d temperature, humidity is 35%, after static dry 24 hours, goes out ball;
F, wash ball: it is the oil stain on 96% ethanol flush away utricule surface that the resulting soft capsule of step e is adopted concentration;
G, choosing are chosen: the soft capsule of the resulting flush away of step f surface oil stain is selected chooses, remove non-eurymeric ball, the eurymeric soft capsule;
H, packing, finished product: with the bottling of the resulting eurymeric soft capsule of step g, seal, finished product.
In the present embodiment, the percentage by weight of the intravital content of soft capsule is respectively: Fructrs Hippophae seed oil 41%, lecithin 1%, vitamin e1 %, lycopene 8%, propolis 9%, Semen Cucurbitae oil 30%, collagen protein 10%; The flexible glue utricule is to be processed in 100: 100: 40 ratio by gelatin, water, glycerol.The soft capsule dosage science of gained of the present invention, defying age, the protection cardiovascular effective, can effectively improve body immunity, and have the supplementing the kidney to control the nocturnal function; Its preparation method technology is reasonable, and is workable, is prone to accomplish scale production.
Adopt animal experiment to explain that soft capsule of the present invention has defying age below
, protection cardiovascular, the function of raising immunity.
1, material and method
1.1, given the test agent:According to soft capsule preparation method provided by the invention, produce 500mg/ grain soft capsule sample.According to the human body recommended intake is every day 2 times, and each 2, promptly human body recommended amounts every day is 2.0g/60kgbw (33.33mg/kgbw).
1.2, experimental animal and component:The healthy SPF Kunming mouse that laboratory animal is provided by Shandong University zoopery center.Laboratory animal is divided into 4 immunity at random organizes greatly, 55 of every big groups.
1.3, the experimental situation condition:Experimental situation is a barrier environment.22 ℃ ± 2 ℃ of ambient temperatures, relative humidity 50% ± 10%.
1.4, dosage is selected with tried thing gives and mode:According to human body recommended amounts (33.33mg/ day/kgbw); Basic, normal, high dosage by 5 times, 10 times, 30 times definite mices that are equivalent to the human body recommended amounts is respectively: 0.17g/kgbw, 0.33g/kgbw, 1g/kgbw; Per os is irritated stomach once a day and is given mice and tried thing, irritates the long-pending 10mL/kgbw that presses of body of stomach.Tried thing with Semen Maydis oil dissolving and dilution before irritating stomach, matched group replaces being tried thing with isopyknic distilled water and Semen Maydis oil respectively, gives continuously to survey each item immune indexes behind the 30d.
1.5, key instrument and reagent
Instrument: animal balance, analytical balance, clean bench, slide gauge (precision 0.01mm), microsyringe (25 μ L), CO
2Incubator, water bath with thermostatic control, centrifuge, ultraviolet spectrophotometer, enzyme-linked immunosorbent assay instrument, optical microscope, micro sample adding appliance (20～200 μ L, 100～1000 μ L).Aseptic operation apparatus, 200 eye mesh screens, glass dish, gauze, test tube, hematimeter, flat Tissue Culture Plate, U type Tissue Culture Plate, timer, hematochrome suction pipe, slide frame.
Reagent: sheep red blood cell (SRBC) (SRBC); Normal saline; Concanavalin A, Con A (ConA); Complement (GPS); Hank ' s liquid; Mycillin; The SA buffer; Agarose; MTT; Calf serum; The PRMI1640 culture fluid; The PBS buffer; Sodium lactate; The nitro tetrazolium chloride; Azophenlyene dimethyl ester phosphate; NAD; 1% glacial acetic acid; 1mol/L HCl; 0.1% NP40 (v/v); Acid isopropyl alcohol (the 96mL isopropyl alcohol adds the HCl of 4mL 1mol/L); The YAC-1 cell; 0.2mol/L the Tris-HCl buffer; India ink; 1g/L Na
2CO
3, chicken red blood cell, methanol, acetone Giemsa dye liquor etc.
1.6, experimental technique.
1.6.1, the organ weight ratio pH-value determination pH
Dislocation was put to death after mice was weighed, and got its thymus, spleen, removed most fascia, blotted the organ surface blood stains and weighed calculating thymus body weight ratio and spleen body weight ratio with filter paper.The gained data are carried out variance analysis with the PEMS statistical software.
1.6.2, delayed allergy (DTH) (the sufficient sole of the foot thickens method)
Get Sanguis caprae seu ovis, de-fibering, with normal saline washing 3 times, every mice is through lumbar injection 2% (v/v prepares with normal saline) hematocrit SRBC 0.2mL, and after the sensitization 4 days, measurement of left foot back sole of the foot portion thickness, same position is measured three times, averages.The subcutaneous injection 20% hematocrit SRBC 20 μ L in the measuring point in injecting back 24h measurement of left metapedes sole of the foot portion thickness, represent the degree of DTH with the difference of sufficient sole of the foot thickness before and after attacking then.
The gained data are calculating chart; Carry out variance analysis with the PEMS statistical software; If tried the thing group attack before and after the difference of sufficient sole of the foot thickness be higher than matched group, and difference has significance (P < 0.05), this is tried decidable thing the effect that improves mice delayed allergy ability is arranged.
1.6.3, the inductive mouse spleen lymphocyte transformation experiment of ConA (mtt assay)
The aseptic spleen of getting places the little plate that fills an amount of aseptic Hank ' s liquid, processes cell suspension.Use Hank ' s liquid washing 2 times, each centrifugal 10min of 1000rpm.Then cell suspension is floated in the complete culture solution of 1mL RPMI1640, the living cell counting number, the adjustment cell concentration is 3 * 10
6Individual/mL.Divide two holes to add in the culture plate cell suspension, every hole 1mL, a hole adds 75 μ L ConA liquid therein, and another hole places 5% CO as contrast
2, 37 ℃ of incubators cultivate 68h, every hole adds 50 μ L MTT, continues to cultivate 4h.Every hole adds 1mL acid isopropyl alcohol, and the piping and druming mixing dissolves purple crystal fully, then liquid is moved in the cuvette, uses ultraviolet spectrophotometer, measures its OD value (OD value) in the 570nm wavelength.Calculate the poor of test hole OD value and control wells OD value, represent lymphocytic multiplication capacity with this.
The gained data are calculating chart; Carry out variance analysis with the PEMS statistical software; If tried the lymphocytic multiplication capacity of thing group and be higher than matched group, and difference has significance (P < 0.05), and this is tried decidable thing the effect that improves the inductive mouse lymphocyte conversion capability of ConA is arranged.
1.6.4, antibody-producting cell measures (Jerne improves slide method)
Get Sanguis caprae seu ovis, de-fibering, with normal saline washing 3 times, every mice is put to death the mice of SRBC immunity after 5 days through lumbar injection 2% (v/v prepares with normal saline) hematocrit SRBC 0.2mL, gets spleen, processes cell suspension.After 10g/L agar heating for dissolving, mix the packing small test tube with the double Hank ' s of equivalent liquid; Every pipe 0.5mL adds 10% (v/v is with the preparation of SA liquid) hematocrit SRBC 50 μ L, splenocyte suspension 25 μ L again in pipe; Behind the mixing, be poured on the slide of brushing the agarose thin layer rapidly, treat that agar solidifies after; The slide level buckled be placed on the horse, put into CO
2Incubator is hatched 1.5h, then complement added in the slide frame groove, and behind the continuation incubation 1.5h, counting hemolysis plaque number.
The gained data are calculating chart, carry out variance analysis with the PEMS statistical software, if tried thing group hemolysis plaque number and be higher than matched group, and difference has significance (P < 0.05), and this is tried the effect that thing has increases the mouse antibodies cell number decidable.
1.6.5, the mensuration of mice serum hemolysin
Get Sanguis caprae seu ovis, de-fibering is with normal saline washing 3 times; Every mice is through lumbar injection 2% (v/v prepares with normal saline) hematocrit SRBC 0.2mL, and immunity was extractd eyeball of mouse after 5 days; Get blood in centrifuge tube, place 1h, solidification blood and tube wall are peeled off; It is centrifugal that (2000rpm 10min), collects serum.Put about 1h with normal saline, solidification blood and tube wall are peeled off, centrifugal (2000rpm 10min), collects serum., with the serum doubling dilution different dilution factor serum are placed respectively in the Microhemagglutination bread board with normal saline, every hole 100 μ L add 0.5% SRBC suspension, 100 μ L again, and mixing places in the wet box, in 37 ℃ of incubation 3h, and record hemagglutination degree.The calculating antibody product.
Serum coagulation degree is divided into 5 grades, by following standard determination.0 grade: SRBC all sinks, and concentrates on the round point shape that sensitization is formed on the bottom, hole, all around liquid clear; The I level: the SRBC major part is deposited on the bottom, hole and forms round point shape, and a small amount of agglutinative SRBC is arranged all around; The II level: agglutinative SRBC forms thin layer in the bottom, hole, and a loose red point can be obviously seen at the center; The III level: the even shakedown of agglutinative SRBC becomes skim at the bottom of being dispersed in the hole, and a red point mays be seen indistinctly at the center; The IV level: agglutinative SRBC becomes skim at the bottom of the hole, grumeleuse becomes the convolution shape sometimes.
The gained data are calculating chart, carry out variance analysis with the PEMS statistical software, if tried thing group serum hemolysin antibody product and be higher than matched group, and difference has significance (P < 0.05), and this is tried the effect that thing has increases the mice serum hemolysin level decidable.
1.6.6, mice carbon cleans up experiment
The india ink of mouse tail vein injection dilution in 1: 5 treats that prepared Chinese ink injects, timing immediately, and 2min, 10min get blood 20 μ L from angular vein respectively behind the injection prepared Chinese ink, and it is added 2mL Na
2CO
3In the solution.With ultraviolet spectrophotometer at 600nm wavelength photometry density value, with Na
2CO
3Solution is made blank.Mice is put to death, get its liver and spleen and weigh.Be calculated as follows phagocytic index a:
K=(lgOD
1– lgOD
2)/(t
2– t
1), a=body weight ÷ (liver weight+spleen is heavy) * k
1/3
The gained data are calculating chart, carry out variance analysis with the PEMS statistical software, if the phagocytic index that is tried the thing group is higher than matched group, and difference has significance (P < 0.05), the effect that this carbon that is tried thing and have increase mouse monokaryon-macrophage of decidable is cleaned up ability.
1.6.7, Turnover of Mouse Peritoneal Macrophages engulfs chicken red blood cell experiment (half intracorporal method)
Mouse peritoneal is injected 20% hematocrit chicken erythrocyte suspension 1mL, and 30min dislocates and puts to death at interval; Get peritoneal macrophage washing liquid 1mL, drip on microscope slide, put into the enamel box that is lined with wet gauze; After moving into 37 ℃ of incubator incubation 30min, rinsing in normal saline is dried.Fix with 1: 1 acetone methanol, the dyeing of 4%Giemsa-phosphate buffer, the rinsing of reuse distilled water is dried.The oil mirror is counting down, and 100 macrophages of every counting are calculated as follows phagocytic rate and phagocytic index:
Phagocytic rate %=engulfs macrophage number * 100 of the macrophage number/counting of chicken red blood cell
The macrophage number of the chicken red blood cell sum/counting that phagocytic index=quilt is engulfed
The gained data are calculating chart, if the phagocytic rate or the phagocytic index that are tried the thing group organize apparently higher than 0.000g/kgbw, and difference has significance (P < 0.05), and this is tried decidable thing and have and increase the effect that mouse macrophage is engulfed the chicken red blood cell ability.
1.6.8, NK cells in mice determination of activity (lactate dehydrogenase L DT algoscopy)
24h is cultivations of going down to posterity of YAC-1 cell before the test, gives a baby a bath on the third day after its birth time with Hank ' s liquid with preceding, and it is 4 * 10 that use RPMI1640 complete culture solution is adjusted cell concentration
5Individual/mL.The mice dislocation is put to death, the aseptic spleen of getting, the preparation splenocyte suspension, it is inferior to use Hank ' s liquid to give a baby a bath on the third day after its birth, each centrifugal 10min of 1000rpm.It is resuspended that reuse 2mL contains the RPMI1640 complete culture solution of 10% calf serum, and with the blue dyeing counting of platform phenol, the adjustment cell concentration is 2 * 10
7Individual/mL, making and imitating the target ratio is 50: 1.
Get each 100 μ L of target cell and splenocyte, add U type culture plate, target cell nature release aperture adds target cell and each 100 μ L of culture fluid, and the maximum release aperture of target cell adds target cell and each 100 μ L of 1%NP40, and above-mentioned each item is all established three parallel holes, 5% CO
2, 37 ℃ of incubators cultivate 4h, with the centrifugal 5min of 1500rpm, every hole is got supernatant 100 μ L and is placed ELISA Plate with culture plate; Add LDH substrate liquid 100 μ L simultaneously; Reaction 3min, every then hole adds the HCl 30 μ L cessation reactions of 1mol/L, measures the OD value at the 490nm place with ELIASA.Be calculated as follows the NK cytoactive:
NK cytoactive %=[(reacting hole OD-nature release aperture OD)/(maximum release aperture OD-nature release aperture OD)] * 100
The preparation of LDH substrate liquid: EINECS 212-761-8 5 * 10
-2Mol/L
Nitro tetrazolium chloride 6.6 * 10
-4Mol/L
Azophenlyene dimethyl ester sulfate 2.8 * 10
-4Mol/L
NAD 1.3 * 10
-3Mol/L
Mentioned reagent is dissolved in the Tris-HCl buffer of 0.2mol/L
The gained data are calculating chart, carry out variance analysis with the PEMS statistical software, and the NK cytoactive need be by X=Sin
-1(P)
1/2Carry out data transaction, P is the NK cytoactive in the formula, and expression decimally be higher than matched group if try the NK cytoactive of thing group, and difference has significance (P<0.05), this is tried the effect that thing has increases the active ability of NK cells in mice decidable.
1.7, experimental data statistics
Experimental data is carried out homogeneity test of variance with the PEMS statistical software to each experiment initial data, satisfies data information that " variance is neat " require and carries out statistical disposition with the comparative approach in twos of mean between a plurality of experimental grouies and matched group; The data information of heterogeneity of variance is carried out suitable variable conversion, after waiting to satisfy " variance is neat " and requiring, carry out statistical disposition with the data of conversion gained.
1.8, the result judges
Any two aspects result is positive aspect four of cellular immune functions, humoral immune function, monokaryon-macrophage function, NK cytoactive, and this given the test agent of decidable has the function of enhancing immunity effect.Wherein two experimental results in the cellular immune function assay project are all positive, and perhaps arbitrary experiment above dose groups result is positive, and decidable cellular immune function assay result is positive.Two experimental results in the humoral immune function mensuration project are all positive, and perhaps arbitrary experiment above dose groups result is positive, and it is positive that the decidable humoral immune function is measured the result.Two experimental results in monokaryon-macrophage function mensuration project are all positive, and perhaps arbitrary experiment above dose groups result is positive, and it is positive that decidable monokaryon-macrophage function is measured the result.An above dose groups result of NK cytoactive experiment is positive, and it is positive that decidable NK cytoactive is measured the result.
2, result
2.1, soft capsule is to the influence of mice body weight:See table 1.
Table 1 soft capsule is to the influence of mice body weight
Visible by table 1, compare between each dose groups of the initial body weight of mice and matched group, there are no significant for difference (P>0.05), promptly the initial body weight of mice is comparatively balanced between each group.Per os gives the soft capsule 30 days of mice various dose, relatively, there are no significant for difference between solvent control group and each dose groups and water matched group for weight gain value (P>0.05), promptly soft capsule does not have influence to the mice body weight.
2.2, soft capsule is to the influence of mice organ weight ratio value:See table 2, table 3.
Table 2 soft capsule is to the influence of mouse spleen body weight ratio
| Group | Number of animals (only) | Spleen body weight ratio (mg/g) | The P value |
| The water matched group | 11 | 4.63±1.26 | ? |
| Solvent control group | 11 | 4.46±1.19 | 0.752 |
| Low dose group | 11 | 4.66±1.01 | 0.948 |
| Middle dose groups | 11 | 4.93±0.93 | 0.522 |
| High dose group | 11 | 5.29±1.20 | 0.218 |
Visible by table 2; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, its spleen body weight ratio between solvent control group and each dose groups and water matched group relatively; There are no significant for difference (P>0.05), promptly soft capsule does not have influence to mouse spleen body weight ratio.
Table 3 soft capsule is to the influence of mouse thymus body weight ratio
| Group | Number of animals (only) | Thymus body weight ratio (mg/g) | The P value |
| The water matched group | 11 | 2.82±0.58 | ? |
| Solvent control group | 11 | 2.99±0.35 | 0.412 |
| Low dose group | 11 | 2.87±0.74 | 0.847 |
| Middle dose groups | 11 | 3.14±0.77 | 0.276 |
| High dose group | 11 | 2.71±0.55 | 0.666 |
Visible by table 3; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, its thymus body weight ratio between solvent control group and each dose groups and water matched group relatively; There are no significant for difference (P>0.05), promptly soft capsule does not have influence to mouse thymus body weight ratio.
2.3, soft capsule is to the mouse cell Immune Effects
2.3.1, soft capsule is to the influence of mice delayed allergy (DTH):See table 4.
Table 4 soft capsule is to the influence of mice delayed allergy (DTH)
| Group | Number of animals (only) | Swelling degree of the paw (mm) | The P value |
| The water matched group | 11 | 0.26±0.07 | ? |
| Solvent control group | 11 | 0.29±0.12 | 0.488 |
| Low dose group | 11 | 0.31±0.14 | 0.288 |
| Middle dose groups | 11 | 0.34±0.12* | 0.033 |
| High dose group | 11 | 0.37±0.13* | 0.021 |
*P<0.05
Visible by table 4; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, its swelling degree of the paw between solvent control group and low dose group and water matched group relatively; There are no significant for difference (P>0.05); Relatively, difference all has significance (P < 0.05) between middle and high dose groups and water matched group, and promptly soft capsule is to strengthening the delayed allergy ability of mice.
2.3.2, soft capsule is to the influence of ConA inducing mouse lymphocyte transformation experiment:See table 5.
Table 5 soft capsule is to the influence of ConA inducing mouse lymphocyte transformation experiment
| Group | Number of animals (only) | Lymphopoiesis ability (OD difference) | The P value |
| The water matched group | 11 | 0.090±0.021 | ? |
| Solvent control group | 11 | 0.082±0.041 | 0.564 |
| Low dose group | 11 | 0.090±0.031 | 0.962 |
| Middle dose groups | 11 | 0.116±0.032* | 0.037 |
| High dose group | 11 | 0.117±0.030* | 0.022 |
*P<0.05
Visible by table 5; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, its lymphopoiesis ability between solvent control group and low dose group and water matched group relatively; There are no significant for difference (P>0.05); Relatively, difference all has significance (P < 0.05) between middle and high dose groups and water matched group, and promptly soft capsule is to strengthening the lymphocyte transformation ability of mice.
2.4, soft capsule is to the influence of mouse humoral immune
2.4.1, soft capsule is to the influence of mouse antibodies cellulation number:See table 6.
Table 6 soft capsule is to the influence of mouse antibodies cellulation number
| Group | Number of animals (only) | Hemolysis plaque number (* 10 3/ full spleen) | The P value |
| The water matched group | 11 | 60.41±18.45 | ? |
| Solvent control group | 11 | 67.16±22.91 | 0.456 |
| Low dose group | 11 | 72.41±21.51 | 0.176 |
| Middle dose groups | 11 | 82.02±22.83* | 0.024 |
| High dose group | 11 | 84.68±25.90* | 0.020 |
*P<0.05
Visible by table 6; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, its antibody-producting cell number between solvent control group and low dose group and water matched group relatively; There are no significant for difference (P>0.05); Relatively, difference all has significance (P < 0.05) between middle and high dose groups and water matched group, and promptly soft capsule is to improving the effect of mouse antibodies cellulation number.
2.4.2, soft capsule is to the influence of mice serum hemolysin:See table 7.
Table 7 soft capsule is to the influence of mice serum hemolysin
| Group | Number of animals (only) | The antibody product | The P value |
| The water matched group | 11 | 176.5±29.3 | ? |
| Solvent control group | 11 | 173.2±28.8 | 0.798 |
| Low dose group | 11 | 180.6±32.6 | 0.755 |
| Middle dose groups | 11 | 202.6±24.0* | 0.033 |
| High dose group | 11 | 204.6±21.6* | 0.018 |
*P<0.05
Visible by table 7; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, between its antibody product solvent control group and low dose group and water matched group relatively; There are no significant for difference (P>0.05); Relatively, difference all has significance (P < 0.05) between middle and high dose groups and water matched group, and promptly soft capsule is to improving the effect of mice serum hemolysin level.
2.5, soft capsule is to the influence of mouse monokaryon-macrophage phagocytic function
2.5.1, soft capsule is to the influence of mouse monokaryon-macrophage phagocytic function:See table 8.
Table 8 soft capsule is to the influence of mouse monokaryon-macrophage phagocytic function
| Group | Number of animals (only) | Phagocytic index | The P value |
| The water matched group | 11 | 4.13±0.73 | ? |
| Solvent control group | 11 | 4.15±0.76 | 0.953 |
| Low dose group | 11 | 4.18±0.84 | 0.887 |
| Middle dose groups | 11 | 4.81±0.66* | 0.032 |
| High dose group | 11 | 5.01±0.79* | 0.013 |
*P<0.05
Visible by table 8; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, its monokaryon-macrophage carbon clean up ability between solvent control group and low dose group and water matched group relatively; There are no significant for difference (P>0.05); Relatively, difference all has significance (P < 0.05) between middle and high dose groups and water matched group, and promptly soft capsule is cleaned up ability to strengthening mouse monokaryon-macrophage carbon.
2.5.2, soft capsule engulfs the influence of chicken red blood cell ability to Turnover of Mouse Peritoneal Macrophages:See table 9, table 10.
Table 9 soft capsule is to the influence of Turnover of Mouse Peritoneal Macrophages chicken red blood cell phagocytic rate
*P<0.05
Table 10 soft capsule is to the influence of Turnover of Mouse Peritoneal Macrophages chicken red blood cell phagocytic index
| Group | Number of animals (only) | Phagocytic index | The P value |
| The water matched group | 11 | 0.50±0.17 | ? |
| Solvent control group | 11 | 0.51±0.16 | 0.828 |
| Low dose group | 11 | 0.59±0.18 | 0.231 |
| Middle dose groups | 11 | 0.67±0.16* | 0.023 |
| High dose group | 11 | 0.69±0.19* | 0.019 |
*P<0.05
Visible by table 9, table 10; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, its peritoneal macrophage chicken red blood cell phagocytic rate and phagocytic index between solvent control group and low dose group and water matched group relatively; There are no significant for difference (P>0.05); Relatively, difference all has significance (P < 0.05) between middle and high dose groups and water matched group, and promptly soft capsule is engulfed the ability of chicken red blood cell to strengthening Turnover of Mouse Peritoneal Macrophages.
2.6, soft capsule is to the active influence of NK cells in mice:See table 11.
Table 11 soft capsule is to the active influence of NK cells in mice
*P<0.05
Visible by table 11; Per os gives the soft capsule 30 days of mice various dose, through statistical procedures, its NK cytoactive between solvent control group and low, middle dose groups and water matched group relatively; There are no significant for difference (P>0.05); Relatively, difference all has significance (P < 0.05) between high dose group and water matched group, and promptly soft capsule is to strengthening the NK cells in mice activity.
3, conclusion
Per os gives the soft capsule 30 days of mice various dose; Weight of mice is had no adverse effects; Mouse spleen body weight ratio and thymus body weight ratio are not had influence, and the cellular immune function of mice, humoral immune function, monokaryon-macrophage phagocytic ability and NK cytoactive functional examination result are all positive.The result shows that the soft capsule that adopts method for preparing provided by the present invention to process has function of enhancing immunity and defying age, protects cardiovascular effect.
Claims (3)
1. a defying age, protection cardiovascular soft capsule; It is made up of flexible glue utricule and the intravital content of this soft capsule of inclosure, and it is characterized in that: described content is made up of following components in weight percentage: Fructrs Hippophae seed oil 40%～60%, lecithin 1%～3%, vitamin e1 %～3%, lycopene 4%～8%, propolis 5%～9%, Semen Cucurbitae oil 20%～30%, collagen protein 6%～10%; Described flexible glue utricule is to be processed in 100: 100: 40 ratio by gelatin, water, glycerol, and the weight ratio of said flexible glue utricule and said content is: 1: 2～3.
2. a kind of defying age according to claim 1, protection cardiovascular soft capsule, it is characterized in that: the percentage by weight of the component of described content is respectively: Fructrs Hippophae seed oil 50%, lecithin 2%, vitamin E2 %, lycopene 6%, propolis 7%, Semen Cucurbitae oil 25%, collagen protein 8%.
3. a kind of defying age according to claim 1, protection cardiovascular preparation of soft capsule method is characterized in that: through following technology,
A, batching: according to percentage by weight weighing Fructrs Hippophae seed oil 40%～60%, lecithin 1%～3%, vitamin e1 %～3%, lycopene 4%～8%, propolis 5%～9%, Semen Cucurbitae oil 20%～30%, collagen protein 6%～10%, place material-compound tank, stir;
B, grinding, the degassing: place colloid mill to grind three times the Fructrs Hippophae seed oil that stirs, lecithin, vitamin e1, lycopene, propolis, Semen Cucurbitae oil, the collagen protein of step a gained; Cross 100 mesh sieves; Place vacuum kettle then; Control vacuum-0.06Mpa sloughs gas, gets the content material;
The making of c, flexible glue envelop materials: gelatin, water, glycerol in ratio weighing in 100: 100: 40, in the being shipped to glue jar, are stirred and are warming up to 60～75 ℃; Treat that gelatin all melts, decompression vacuum pumping, control vacuum-0.06Mpa; Slough gas, cross 80 mesh sieves, obtain flexible glue utricule filtrating; Contain in heat-preserving container, subsequent use;
D, pelleting: with the resulting flexible glue utricule of step c filtrating and the resulting content material of step b according to weight ratio: 1: 2～3 ratio is sent in the pellet press, suppresses soft capsule;
E, typing drying: with 22～26 ℃ of the resulting soft capsule control of steps d temperature, humidity is 30～40%, after static dry 24 hours, goes out ball;
F, wash ball: it is the surperficial oil stain of ethanol flush away utricule 90% or more that the resulting soft capsule of step e is adopted concentration;
G, choosing are chosen: the soft capsule of the resulting flush away of step f surface oil stain is selected chooses, remove non-eurymeric ball, the eurymeric soft capsule;
H, packing, finished product: with the bottling of the resulting eurymeric soft capsule of step g, seal, finished product.
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