CN105770066A - Probiotic fermented traditional Chinese medicine compound preparation and preparation method and application thereof - Google Patents
Probiotic fermented traditional Chinese medicine compound preparation and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a probiotic fermented traditional Chinese medicine compound preparation and a preparation method thereof.The compound is prepared from cassia seeds, chrysanthemum flowers, the root of kudzu vine, hawthorn fruits and fiveleaf gynostemma herb through water extraction, concentration, enzymolysis, adding of a carbon source, a nitrogen source and inorganic salt, sterilization and inoculation of probiotics for fermentation.The compound can be used in medicine or health care products for improving body immunity and preventing and treating hyperlipemia.Test results show that the probiotic fermented traditional Chinese medicine compound preparation can significantly lower the concentration of total cholesterol and triglyceride of rat, has the effect of adjusting blood lipids, can significantly increase the number of mouse antibody generation splenocytes and significantly improve the mouse macrophage phagocytic function, and has the immunity regulating function.
Description
Technical field
The present invention relates to a kind of for blood fat reducing and the symbiotically fermented middle medicine composite preparation of many probiotics improving body immunity and preparation method thereof, belong to pharmaceutical technology field.
Background technology
Cardiovascular and cerebrovascular disease becomes No. 1 in the world underlying cause of death, and its prevalence is the trend risen year by year with mortality rate, and the raising of serum cholesterol level is the principal element causing coronary heart disease.Reduce serum cholesterol level and be directly connected to the health of mankind itself.Hyperlipidemia is chronic disease, there is no any external symptom, only noiseless consequence " arteriosclerosis ", by the time when blood vessel blockage more than 70%, patient just can experience symptom because hyperlipidemia lacks symptom, once morbidity can bring again serious cardiovascular and cerebrovascular disease, hyperlipidemia is referred to as noiseless killer.
Regulate the content of human serum cholesterol and triglyceride, prevent the countermeasure of hyperlipemia mostly around reducing its intake, suppress self synthesis and promote that converting excretion these three aspect launches, sum up and mainly have five kinds of methods: (1) is by the autologous synthesis of drugs block cholesterol and triglyceride. Monacolin, lovastatin, the anticholesteremic agents such as pravastatin are through suppressing the activity of the key enzyme (3-base 1 monomethyl penta 2 phthalein CoA reductase) in cholesterol biosynthesis approach to reduce cholesterol self synthesis and induced liver specific fat acid transporter albumen and the expression of second phthalein CoA synthase, promote liver picked-up fatty acid, liver synthetic glycerine three ester is suppressed to reduce serum cholesterol and content of triglyceride.(2) small intestinal reuptaking cholate is suppressed, cholesterol is the body important channel to external excretion of cholesterol to the conversion of cholic acid, also it is the crucial point of adjustment affecting blood lipid level, suppress reuptaking of cholic acid in intestinal that the content of cholic acid in liver can be made to decline, more cholesterol is promoted to be converted into cholic acid in liver, thus reducing the content of serum cholesterol.Cellulose, medicine, some lactic acid bacteria etc. all can suppress cholic acid in intestinal to reuptake, interference hepato-enteric circulation, thus reducing serum cholesterol concentration: the edible cholesterol of (3) restriction and the high food of content of triglyceride, with the absorption to dietary cholesterol and triglyceride of the competitive inhibition human body.(4) cholesterol in food and triglyceride are removed by physics, chemical method;(5) biotransformation method removes the cholesterol in food and triglyceride, the application in food of this method can have two kinds of approach: a kind of be with microorganism directly assimilate, the cholesterol of degrading in food and triglyceride, another kind is that the enzymes such as the oxidase, the reductase that microorganism are produced are applied to food.
Body's cholesterol source mainly has two kinds: one is endogenous synthesis, and two is external source picked-up, and therefore, edible blood fat reducing food, reduces the intake of cholesterol in daily life, is the effective means reducing serum cholesterol content.Human body probiotic bacteria, except having the effect regulating human body intestinal canal flora, maintaining human body microecological balance, also has the effect reducing human serum cholesterol.Research at present has three kinds of mechanism to reduce the content of cholesterol, wherein probiotic bacteria effect for reducing blood fat is probably cholesterol and the Mycoderma compact siro spinning technology of absorption, it is likely to penetrate into or be integrated into after birth, cholesterol can be adsorbed in cell surface by thalline under cholate existent condition, and this reaction requires the existence of cholate in environment.Probiotic bacteria blood fat reducing, firstly the need of cholesterol molecule is adsorbed in Membranes surface, then could be penetrated into or be incorporated into cell membrane.
Reducing Blood Lipid by Chinese medicine complicated mechanism, plays comprehensive function often through multicomponent, multipath.Its main path has the synthesis reducing endogenous lipid, reduce exogenous cholesterol absorption, promote the operating of lipid and row, regulating lipid metabolism etc..Make a general survey of present Research in recent years, Chinese medicine treatment hyperlipemia, prevent cardiovascular and cerebrovascular disease generation etc. in play the advantage of uniqueness, its curative effect certainly, continue, few side effects, be that Western medicine institute is incomparable.
The instructions of taking of Chinese medicine mostly generally is oral.With the Chinese medicine that decoction form is oral, its chemical composition is then directly entered digestive tract.The behavior in digestive tract of this alien material chemical composition includes: 1. with prototype compound form absorbed into serum；2. with digestive tract bioconversion and (or) metabolite form absorbed into serum；3. directly excrete with feces with prototype or bioconversion thing and (or) metabolite form；4. the wall that stimulates digestion causes immune activation to produce biological effect；5. enteral Tiny ecosystem is stimulated to cause bioactive substance secretion to produce biological effect；6. destroy enteral microecological balance and promote that new balance system is set up.
Intestinal bacterium is of crucial importance to Chinese medicine ingredients effect.Human body intestinal canal is the habitat of a huge and dynamic bacteria flora, human intestine there are about 100,000,000 antibacterials, more than 99% is anaerobe, and wherein the obligate anaerobe such as Bacteroideae (Bacteriodacae), catenabacterium (Cattenabacterium), peptostreptococcus (Peptostreptococcus), Spirillum (Spirillaceal) and lactic acid bacteria and bacillus bifidus are preponderated.Intestinal microbial population can produce many drug metabolism enzymes, and different types of antibacterial produces different metabolic enzymes, and different drug metabolism enzymes participates in again in different types of drug metabolism.These antibacterials can pass through " pharmacological activation " makes inactive medicine be converted into activated medicine, makes the material Non-toxic being harmful to simultaneously, and vice versa.Interact performance and the pipe intestinal digesting systemic disease to Study of Traditional Chinese Medicine drug effect of Chinese medicine and intestinal microbial population have great importance.
Summary of the invention
It is an object of the present invention to provide medicine composite preparation in a kind of probiotics fermention, technical scheme is as follows:
Choosing study of semen cassia, Flos Chrysanthemi, Radix Puerariae, Fructus Crataegi, Herb Gynostemmae Pentaphylli, carry through water, concentrate, enzymolysis, add carbon source, nitrogenous source and inorganic salt, after sterilizing, inoculation probiotic bacteria is made after fermenting.
Described Chinese medicine content (weight g/ volume ml) in compound formulation is: Semen Cassiae 5-15%, Flos Chrysanthemi 3.0-5.0%, Radix Puerariae 8-20%, Fructus Crataegi 8-15%, Herb Gynostemmae Pentaphylli 1-5%.
Wherein, a preferred scheme is: Semen Cassiae 10%, Flos Chrysanthemi 3%, Radix Puerariae 15%, Fructus Crataegi 10%, Herb Gynostemmae Pentaphylli 1%.
Semen Cassiae is leguminous plant CassiaobtusifoliaL. or the dry mature seed of little Semen Cassiae CassiatoraL.Sweet, bitter, salty, be slightly cold.Return liver, large intestine channel.There is blood pressure lowering, blood fat reducing and antibacterial effect.
Flos Chrysanthemi is the dry capitulum of feverfew chrysanthemum ChrysanthemummorifoliumRamat..Sweet, bitter, it is slightly cold.Return lung, Liver Channel.There is scattered wind heat clearing away, the effect of suppressing the hyperactive liver improving eyesight.It is clinically used for treatment coronary heart disease, hypertension.
Radix Puerariae is legume pueraria lobata Puerarialobata(Willd.) dry root of Ohwi or Radix Puerariae rattan PuerariathomsoniiBenth..Property is sweet, pungent, cool.Return spleen, stomach warp.There is expelling pathogenic factors from muscles for reducing heat, promote the production of body fluid, rash, yang invigorating anti-diarrhea effect.It is clinically used for treatment coronary heart disease.
Fructus Crataegi is the dry mature fruit of rosaceous plant Fructus Pyri Pashiae CrataeguspinnatifidaBge.var.majorN.E.Br. or Fructus Crataegi CrataeguspinnatifidaBge..Sour, sweet, the tepor of property.Return spleen, stomach, Liver Channel.There is promoting digestion and invigorating the stomach, the effect of circulation of qi promoting dissipating blood stasis.It is clinically used for blood pressure lowering, reduces cholesterol and antioxidation.
Cucurbitaceae genus gynostemma Herb Gynostemmae Pentaphylli Gynostemmapentaphyllum(Thunb.) Makino, is used as medicine with root stock.Property bitter, cold, there is heat-clearing and toxic substances removing, cough-suppressing phlegm-dispelling functions.It is clinically used for treatment hyperlipemia, improves immunity, defying age.
In order to improve preparation stability and palatability, Semen Cassiae water precipitate with ethanol again after carrying, alcohol precipitation concentration is 60%.
For the apparent condition of preparation, after Semen Cassiae extracting solution mixes with Flos Chrysanthemi, Radix Puerariae, Fructus Crataegi, Herb Gynostemmae Pentaphylli extract, select pectinase enzymatic hydrolysis, to remove the pectin in Chinese medicine extract.
The probiotic bacteria used in Chinese medicine extract fermentation is selected from bacillus bifidus, lactobacillus, streptococcus.Described bacillus bifidus is selected from bifidobacterium bifidum, bifidobacterium breve, bifidobacterium adolescentis and bifidobacteria infantis；Described lactobacillus selected from bacillus acidophilus, Deshi Lactobacillus, lactobacillus casei and lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus fermenti, described streptococcus selected from streptococcus thermophilus (strain derive from China General Microbiological culture presevation administrative center (CGMCC) (China, Beijing) or separate from the feces or intestinal submucosa tissue of normal person, in condensed milk obtain these bacterial isolateses).
According to a preferred embodiment of the invention, described bacillus bifidus is selected from bifidobacterium adolescentis and bifidobacteria infantis；Described lactobacillus is selected from Deshi Lactobacillus, Lactobacillus fermenti and bacillus acidophilus, and streptococcus is selected from streptococcus thermophilus.
It is a further object to provide the preparation method of medicine composite preparation in above-mentioned probiotics fermention, step is as follows: Semen Cassiae adds the water of 6 ~ 8 times, soaks 30 ~ 60 minutes, decoct twice, each 1 hour, merge decoction liquor, filter and concentrating under reduced pressure, making liquor strength is 50%(medical material weight g/ volume ml), add ethanol, making concentration of alcohol is 60%, stand overnight, filter, reclaim and wave most ethanol, obtain Semen Cassiae extracting solution.Flos Chrysanthemi, Radix Puerariae, Fructus Crataegi, Herb Gynostemmae Pentaphylli add the water of 6 ~ 8 times, soak 30 ~ 60 minutes, decoct twice, each 1 hour, merge decoction liquor, and filtration is concentrating under reduced pressure also, merges with Semen Cassiae extracting solution, and making containing medical material concentration is 25 ~ 60%(medical material weight g/ volume ml).Mixed extracting solution is adjusted temperature is 50 DEG C, pectin lyase is added according to millesimal ratio (volume/volume), enzyme liquid drinking water is diluted to the concentration use of 10%, after stirring, enzymolysis 2 hours, after terminating enzymolysis, 100 DEG C of enzyme denaturing 10 minutes, it is centrifuged or filters, take clear liquid, obtain Semen Cassiae, Flos Chrysanthemi, Radix Puerariae, Fructus Crataegi, Herb Gynostemmae Pentaphylli Chinese medicine extraction liquid.Add carbon source, nitrogenous source and inorganic salt, sweeting agent, adjust pH value to neutral, after high temperature sterilize, obtain probiotics fermention substrate；Inoculating pre-incubated is respectively 0.5%(volume/volume) at least one bacillus bifidus bacterium solution, after cellar culture about 2 ~ 6 hours, again inoculate pre-incubated respectively for 0.5%(volume/volume) lactobacillus, at least one bacterium solution in streptococcus, continue fermentation 20 ~ 22 hours, when pH value is down to below 4.0, terminate fermentation, obtain probiotic bacteria fermented tcm compound formulation.
Described carbon source is glucose, content 1.5%(weight g/ volume ml).
Described nitrogenous source is peptone, content 0.4%(weight g/ volume ml), yeast extract, content 0.2%(weight g/ volume ml).
Described inorganic salt is: K2HPO4·3H2O0.1g/100ml、KH2PO40.1g/100ml、FeSO4·7H2O5.0mg/100ml、ZnSO4·7H2O4.4mg/100ml、MgSO4·7H2O50mg/100ml、Na2SeO3·5H2O0.1mg/100ml。
Described sweeting agent is sucralose, and content is 0.001g/100ml.
The further object of invention is to provide in the probiotics fermention of the present invention medicine composite preparation in preparation and improves application in the medicine of immunity of organisms, prevention and treatment hyperlipemia or health product.
For the raising immunity of medicine composite preparation in the probiotics fermention of the detection present invention and the action effect regulating blood fat, the Immune Function of the experimental observation impact on rat every blood lipids index that high lipid food is fed and mice, test result to show, in the probiotics fermention of the present invention, medicine composite preparation can make rat T-CHOL, triglyceride concentration be remarkably decreased, and has the effect regulating blood fat；In probiotics fermention, medicine composite preparation compares with the Chinese medicine extract without probiotics fermention, it is possible to dramatically increases mouse antibodies and generates spleens cell number, significantly improves macrophage phagocytosis of mice, have immunoloregulation function.
Detailed description of the invention
The invention provides the middle medicine composite preparation of a kind of many probiotics symbiotic fermentation Semen Cassiae, Flos Chrysanthemi, Radix Puerariae, Fructus Crataegi, Herb Gynostemmae Pentaphylli.Many strains are utilized to have the beneficial bacteria of intestinal tract reducing cholesterol effect, in particular with bacillus bifidus and lactobacillus, streptococcus symbiotic fermentation Semen Cassiae, Flos Chrysanthemi, Radix Puerariae, Fructus Crataegi, Herb Gynostemmae Pentaphylli Chinese medicine compound, making medicine composite preparation in probiotics fermention, in this probiotics fermention, medicine composite preparation can be used for preventing and treat in health product or the medicine of hyperlipidemia.
Present invention screening has the probiotics strain of absorption cholesterol effect, the bacterial strain of exploitation probiotic bacteria cholesterol reducing, expands potential range of application and the field of probiotic bacteria.In formula, select the Chinese medicine with blood fat reducing, raising immunity simultaneously, cooperatively play the effect of its blood fat reducing with probiotic bacteria.
Embodiment 1: the preparation of medicine composite preparation A in probiotics fermention.
Press column weight amount and weigh each taste Chinese medicine: Semen Cassiae 100g, Flos Chrysanthemi 30g, Radix Puerariae 150g, Fructus Crataegi 100g, Herb Gynostemmae Pentaphylli 10g.Depend on pine torch, add the water of 6 ~ 8 times, soak 30 ~ 60 minutes, decoct twice, each 1 hour.Merging decoction liquor, filtration, being evaporated to Semen Cassiae concentration is 50%(medical material weight g/ volume ml), add ethanol, make concentration of alcohol reach 60%, after stirring overnight, filter, collect filtrate, reclaim and wave most ethanol, obtain Semen Cassiae extracting solution.Other four Chinese medicines add the water of 6 ~ 8 times, soak 30 ~ 60 minutes, decoct twice, each 1 hour.Merge decoction liquor, filter and concentrating under reduced pressure, add the extracting solution of Semen Cassiae, be finally settled to 1000ml.5 taste Chinese medicine extraction liquid are adjusted temperature is 50 DEG C, and adjusting pH value is 6.0, adds the pectin lyase of Novi's letter according to millesimal ratio (volume/volume), and enzyme liquid drinking water is diluted to the concentration use of 10%.After stirring, enzymolysis 2 hours.Terminate after enzymolysis, 100 DEG C of enzyme denaturing 10 minutes, centrifugal or filter, take clear liquid, obtain Chinese medicine extraction liquid.Add sucralose 0.01g, glucose 15g, yeast extract 2.0g, peptone 4.0g, potassium dihydrogen phosphate 1.0g, dipotassium hydrogen phosphate 3H2O1.0g, magnesium sulfate 7H2O0.5g, ferrous sulfate 7H2O0.05g zinc sulfate 7H2O0.044g, sodium selenite 5H2O1mg, adjusts pH7.0, heats sterilizing in 40 minutes at 115 DEG C.When temperature is down to 39 DEG C, accessing the good bifidobacterium adolescentis of preculture and bifidobacteria infantis bacterium solution, inoculum concentration is 0.5%(volume/volume).After fermenting 6 hours, then accessing pre-incubated Deshi Lactobacillus, bacillus acidophilus's bacterium solution, Lactobacillus fermenti, inoculum concentration is 0.5%(volume/volume), fermentation culture temperature is about 37 DEG C, continues fermentation 20 ~ 22 hours.When pH value is down to below 4.0, terminating fermentation, 115 DEG C inactivate 20 minutes, namely obtain medicine composite preparation A in probiotics fermention.
Embodiment 2: the preparation of medicine composite preparation B in probiotics fermention
Press column weight amount and weigh each taste Chinese medicine: Semen Cassiae 150g, Flos Chrysanthemi 40g, Radix Puerariae 80g, Fructus Crataegi 140g, Herb Gynostemmae Pentaphylli 20g.Depend on pine torch, add the water of 6 ~ 8 times, soak 30 ~ 60 minutes, decoct twice, each 1 hour.Merging decoction liquor, filtration, being evaporated to Semen Cassiae concentration is 50%(medical material weight g/ volume ml), add ethanol, make concentration of alcohol reach 60%, after stirring overnight, filter, collect filtrate, reclaim and wave most ethanol, obtain Semen Cassiae extracting solution.Other four Chinese medicines add the water of 6 ~ 8 times, soak 30 ~ 60 minutes, decoct twice, each 1 hour.Merge decoction liquor, filter and concentrating under reduced pressure, add the extracting solution of Semen Cassiae, be finally settled to 1000ml.5 taste Chinese medicine extraction liquid are adjusted temperature is 50 DEG C, and adjusting pH value is 6.0, adds the pectin lyase of Novi's letter according to millesimal ratio (volume/volume), and enzyme liquid drinking water is diluted to the concentration use of 10%.After stirring, enzymolysis 2 hours.Terminate after enzymolysis, 100 DEG C of enzyme denaturing 10 minutes, centrifugal or filter, take clear liquid, obtain Chinese medicine extraction liquid.Add sucralose 0.01g, glucose 15g, yeast extract 2.0g, peptone 4.0g, potassium dihydrogen phosphate 1.0g, dipotassium hydrogen phosphate 3H2O1.0g, magnesium sulfate 7H2O0.5g, ferrous sulfate 7H2O0.05g zinc sulfate 7H2O0.044g, sodium selenite 5H2O1mg, adjusts pH7.0, heats sterilizing in 40 minutes at 115 DEG C.When temperature is down to 39 DEG C, accessing the good bifidobacterium breve of preculture and bifidobacteria infantis bacterium solution, inoculum concentration is 0.5%(volume/volume).After fermenting 6 hours, then access pre-incubated Lactobacillus plantarum, streptococcus thermophilus, each 0.5%(volume/volume of Deshi Lactobacillus bacterium bacterium solution).Fermentation culture temperature is about 37 DEG C, continues fermentation 20 ~ 22 hours.When pH value is down to below 4.0, terminating fermentation, 115 DEG C inactivate 20 minutes, namely obtain medicine composite preparation B in probiotics fermention.
Embodiment 3: the preparation of medicine composite preparation C in probiotics fermention
Press column weight amount and weigh each taste Chinese medicine: Semen Cassiae 70g, Flos Chrysanthemi 50g, Radix Puerariae 180g, Fructus Crataegi 100g, Herb Gynostemmae Pentaphylli 40g.Depend on pine torch, add the water of 6 ~ 8 times, soak 30 ~ 60 minutes, decoct twice, each 1 hour.Merging decoction liquor, filtration, being evaporated to Semen Cassiae concentration is 50%(medical material weight g/ volume ml), add ethanol, make concentration of alcohol reach 60%, after stirring overnight, filter, collect filtrate, reclaim and wave most ethanol, obtain Semen Cassiae extracting solution.Other four Chinese medicines add the water of 6 ~ 8 times, soak 30 ~ 60 minutes, decoct twice, each 1 hour.Merge decoction liquor, filter and concentrating under reduced pressure, add the extracting solution of Semen Cassiae, be finally settled to 1000ml.5 taste Chinese medicine extraction liquid are adjusted temperature is 50 DEG C, and adjusting pH value is 6.0, adds the pectin lyase of Novi's letter according to millesimal ratio (volume/volume), and enzyme liquid drinking water is diluted to the concentration use of 10%.After stirring, enzymolysis 2 hours.Terminate after enzymolysis, 100 DEG C of enzyme denaturing 10 minutes, centrifugal or filter, take clear liquid, obtain Chinese medicine extraction liquid.Add sucralose 0.01g, glucose 15g, yeast extract 2.0g, peptone 4.0g, potassium dihydrogen phosphate 1.0g, dipotassium hydrogen phosphate 3H2O1.0g, magnesium sulfate 7H2O0.5g, ferrous sulfate 7H2O0.05g zinc sulfate 7H2O0.044g, sodium selenite 5H2O1mg, adjusts pH7.0, heats sterilizing in 40 minutes at 115 DEG C.When temperature is down to 39 DEG C, accessing the good bifidobacterium bifidum of preculture and bifidobacteria infantis bacterium solution, inoculum concentration is 0.5%(volume/volume).After fermenting 6 hours, then access pre-incubated lactobacillus casei, each 0.5%(volume/volume of lactobacillus rhamnosus bacterium solution).Fermentation culture temperature is about 37 DEG C, continues fermentation 20 ~ 22 hours.When pH value is down to below 4.0, terminating fermentation, 115 DEG C inactivate 20 minutes, namely obtain medicine composite preparation C in probiotics fermention.
Experimental example 1: in probiotics fermention, medicine composite preparation regulates blood fat zoopery.
Select sample prepared by embodiment.
Experimental animal is two grades of Wister male rats, 48, weight range 160 ~ 180 grams, medicine composite preparation in probiotics fermention is given in gavage mode, preparation is concentrated 5 times test, each dosage group: matched group 0.0ml/kg.bw, low dose group 3.0ml/kg.bw, middle dosage group 6.0ml/kg.bw, high dose group 9.0ml/kg.bw, the dosage that each dosage group gives tested material is equivalent to 10,20,30 times of people's recommended amounts (0.3ml/kg.bw).In 28 days observation cycles, take fasting 12 hours before blood every time.
Experimental technique: with after normal feedstuff feed rat 5 days, fasting, weigh, take tail blood, enzymatic assays serum total cholesterol (TC), triglyceride (TG), HDL-C (HDL-C).According to body weight, TC level, animal is randomly divided into 4 groups: matched group (0.0ml/kg.bw), three tested material groups (3.0ml/kg.bw, 6.0ml/kg.bw, 9.0ml/kg.bw).From formal test, each treated animal uses high lipid food instead, and tested material prepares desired concn with distilled water.Giving tested material in gavage mode, matched group gives distilled water.Weigh when the 14th, 28 days, take the every blood lipids index of tail hematometry.Experimental data is added up: adopt the significant difference of t-method of inspection inspection experimental group and matched group indices.Experimental result is as follows:
1, the medicine composite preparation impact on rat body weight in probiotics fermention
Before table 1 experiment, in, respectively organize the body weight (g, M ± SD) of rat latter stage
Whole experiment periods, each group Growth in Rats is normal.From table 1, each experimental group rat is compared with matched group, without significant difference (p > 0.05).
2, the medicine composite preparation impact on Serum TC concentration in probiotics fermention
Table 2 experiment before, in, latter stage respectively organize Serum TC concentration (mmol/L, M ± SD)
From table 2, testing latter stage, compared with matched group, sample A, B, C high dose group Serum TC concentration significantly reduces, and has statistically difference (p < 0.05).
3, the medicine composite preparation impact on rat blood serum TG concentration in probiotics fermention
Table 3 experiment before, in, latter stage respectively organize rat blood serum TG concentration (mmol/L, M ± SD)
From table 3, in an experiment, latter stage, compared with matched group, sample A, B, C high dose group rat blood serum TG concentration significantly reduces, and has statistically difference (p < 0.05).
This experimental example shows, compared with the control rats feeding high lipid food, in probiotics fermention, medicine composite preparation can make rat blood serum T-CHOL (TC), triglyceride (TG) concentration be remarkably decreased, and have significant difference (p < 0.05) statistically, therefore in probiotics fermention, medicine composite preparation has the effect regulating blood fat.
Experimental example 2: the experiment that in probiotics fermention, immune function of mice is regulated by medicine composite preparation and unfermentable Chinese medicine extract.
After detecting probiotics fermention, Chinese medicine extract is affected, tested medicine composite preparation and the impact on the immunologic function of mice of the unfermentable Chinese medicine extract in the probiotics fermention of embodiment 1 preparation.
Laboratory animal selects two grades of healthy BALB/C female mices, body weight 18-22.Calculate according to human body recommended amounts everyone every day of 90ml, by two sample concentration one times, design high, medium and low three dosage groups, be respectively equivalent to 30 times of human body recommended amounts, 10 times, 3 times.Matched group directly gives tap water, after giving tested material continuously 15 days, lives and kills mice and measure every immune indexes.Concrete grammar performs according to " health food inspection pre assessment technical specification ".Experimental project includes the experiment of mice carbonic clearance, Turnover of Mouse Peritoneal Macrophages phagocytosis chicken red blood cell experiment (half intracorporal method) and antibody-producting cell detection (Jerne improves slide method).
Experimental data method of analysis of variance is added up.Experimental result is as follows.
1, mouse antibodies is generated the impact of spleens cell number by laboratory sample
Specific experiment result is in Table 4.
Mouse antibodies is generated the impact of spleens cell number by table 4 laboratory sample
From table 4, per os gave the laboratory sample of mice various dose after 15 days, medicine composite preparation middle and high dosage group in probiotics fermention, non-fermented tcm extract high dose group mice hemolysis plaque number have compared with matched group significant difference (*P < 0.05), three dosage groups of probiotic bacteria Chinese medicine composition compare with three dosage groups of not fermenting, it is possible to significantly improve mouse antibodies and generate number, (#P < 0.05), probiotics fermention Chinese medicine is described, it is possible to improve effect of Chinese medicine composition.
2, the laboratory sample impact on mice carbon clearance
Specific experiment result is in Table 5.
The impact on mice carbon clearance of table 5 laboratory sample
From table 5, per os gave the laboratory sample of mice various dose after 15 days, medicine composite preparation basic, normal, high dosage group in probiotics fermention, non-fermented tcm extract high dose group mice phagocytic index have compared with matched group significant difference (*P < 0.05), probiotic bacteria Chinese medicine composition middle and high dosage group compares with middle and high dosage group of not fermenting, it is possible to significantly improve mice phagocytic index, (#P < 0.05), probiotics fermention Chinese medicine is described, it is possible to improve effect of Chinese medicine composition.
3, laboratory sample is the impact on Phagocytosis By The Peritoneal Macrophages In Mice
Specific experiment result is in Table 6.
Table 6 laboratory sample is the impact on Phagocytosis By The Peritoneal Macrophages In Mice
From table 6, per os gave the laboratory sample of mice various dose after 15 days, in probiotics fermention the phagocytic rate of the middle and high dosage group mice of medicine composite preparation have compared with matched group significant difference (*P < 0.05), probiotic bacteria Chinese medicine composition high dose group compares with high dose group of not fermenting, it is possible to significantly improve mice phagocytic rate, (#P < 0.05), probiotics fermention Chinese medicine is described, it is possible to improve effect of Chinese medicine composition.
Claims (10)
1. medicine composite preparation in a probiotics fermention, it is characterised in that choose study of semen cassia, Flos Chrysanthemi, Radix Puerariae, Fructus Crataegi, Herb Gynostemmae Pentaphylli, carry through water, concentrate, enzymolysis, adding carbon source, nitrogenous source, inorganic salt and sweeting agent, after sterilizing, inoculation probiotics bacterial liquid is made after fermenting；
Described Chinese medicine weight g/ volume ml ratio in compound formulation is: Semen Cassiae 5-15%, Flos Chrysanthemi 3.0-5.0%, Radix Puerariae 8-20%, Fructus Crataegi 8-15%, Herb Gynostemmae Pentaphylli 1-5%；
Described probiotic bacteria is selected from bacillus bifidus, lactobacillus or streptococcus.
2. medicine composite preparation in probiotics fermention according to claim 1, it is characterised in that described Chinese medicine weight g/ volume ml ratio in compound formulation is: Semen Cassiae 10%, Flos Chrysanthemi 3%, Radix Puerariae 15%, Fructus Crataegi 10%, Herb Gynostemmae Pentaphylli 1%.
3. medicine composite preparation in probiotics fermention according to claim 1, it is characterised in that described enzymolysis uses pectin lyase.
4. medicine composite preparation in probiotics fermention according to claim 1, it is characterised in that described carbon source is glucose, its weight g/ volume ml ratio is 1.5%；Described nitrogenous source is: peptone and yeast extract, and the weight g/ volume ml ratio of peptone is 0.4%；The weight g/ volume ml ratio of yeast extract is 0.2%.
5. medicine composite preparation in probiotics fermention according to claim 1, it is characterised in that described sweeting agent is sucralose, content is 0.001g/100ml.
6. medicine composite preparation in probiotics fermention according to claim 1, it is characterised in that described inorganic salt is: K2HPO4·3H2O0.1g/100ml、KH2PO40.1g/100ml、FeSO4·7H2O5.0mg/100ml、ZnSO4·7H2O4.4mg/100ml、MgSO4·7H2O50mg/100ml、Na2SeO3·5H2O0.1mg/100ml。
7. medicine composite preparation in probiotics fermention according to claim 1, it is characterised in that described bacillus bifidus is selected from bifidobacterium bifidum, bifidobacterium breve, bifidobacterium adolescentis or bifidobacteria infantis；Described lactobacillus is selected from bacillus acidophilus, Deshi Lactobacillus, lactobacillus casei, lactobacillus rhamnosus, Lactobacillus plantarum or Lactobacillus fermenti；Described streptococcus is selected from streptococcus thermophilus.
8. medicine composite preparation in probiotics fermention according to claim 1, it is characterised in that described bacillus bifidus is selected from bifidobacterium adolescentis and bifidobacteria infantis；Described lactobacillus is selected from Deshi Lactobacillus, Lactobacillus fermenti and bacillus acidophilus.
9. the preparation method of medicine composite preparation in the probiotics fermention described in claim 1-7, it is characterised in that step is as follows:
(1) Semen Cassiae is added the water of 6 ~ 8 times, soaks 30 ~ 60 minutes, decoct twice, each 1 hour, merging decoction liquor, filter and concentrating under reduced pressure, making liquor strength is weight g/ volume ml ratio 50%, add ethanol, making concentration of alcohol is 60%, stands overnight, filters, reclaim and wave most ethanol, obtain Semen Cassiae extracting solution；
(2) Flos Chrysanthemi, Radix Puerariae, Fructus Crataegi, Herb Gynostemmae Pentaphylli being added the water of 6 ~ 8 times, soaks 30 ~ 60 minutes, decocts twice, each 1 hour, merge decoction liquor, filtration is concentrating under reduced pressure also, obtains the extracting solution of four kinds of Chinese medicines；
(3) step (1) is merged with step (2) gained extracting solution, making the strength by weight g/ volume ml ratio containing medical material is 25 ~ 60%, and adjusting temperature is 50 DEG C, adds pectin lyase according to the millesimal ratio of volume ratio, enzyme liquid drinking water is diluted to the concentration use of 10%, after stirring, enzymolysis 2 hours, after terminating enzymolysis, 100 DEG C of enzyme denaturing 10 minutes, it is centrifuged or filters, take clear liquid, obtain five kinds of Chinese medicine extraction liquid；
(4) add carbon source, nitrogenous source, inorganic salt and sweeting agent, adjust pH value to neutral, after high temperature sterilize, obtain probiotics fermention substrate；
(5) inoculating pre-incubated volume ratio is respectively at least one bacillus bifidus bacterium solution of 0.5%, after cellar culture about 2 ~ 6 hours, again inoculating pre-incubated volume ratio is respectively lactobacillus, at least one bacterium solution in streptococcus of 0.5%, continue fermentation 20 ~ 22 hours, when pH value is down to below 4.0, terminate fermentation, obtain probiotic bacteria fermented tcm compound formulation.
10. medicine composite preparation application in the medicine or health product of preparation raising immunity of organisms, prevention and treatment hyperlipidemia in the probiotics fermention described in claim 1-8.
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