CN105749099B - A kind of probiotics fermention Chinese medicine composition and its preparation method and application for treating gastric cancer - Google Patents

A kind of probiotics fermention Chinese medicine composition and its preparation method and application for treating gastric cancer Download PDF

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CN105749099B
CN105749099B CN201610189936.2A CN201610189936A CN105749099B CN 105749099 B CN105749099 B CN 105749099B CN 201610189936 A CN201610189936 A CN 201610189936A CN 105749099 B CN105749099 B CN 105749099B
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CN105749099A (en
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吴炳新
陆继臣
孙筱林
牛纪江
褚新红
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SANZHU fur PHARMACEUTICAL Co Ltd
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Abstract

The invention discloses a kind of probiotics fermention Chinese medicine compositions and its preparation method and application for treating gastric cancer, the fermented tcm composition is by American Ginseng, radix bupleuri, Fructus Aurantii, radix scutellariae, the tuber of pinellia, salvia chinensis, sea Piao sheath, fritillaria thunbergii, Radix Paeoniae Alba, Rhizoma Chuanxiong, prepared RHIZOMA CYPERI with vinegar, galangal, Radix Angelicae Sinensis, Herba Cistanches, radix glycyrrhizae preparata, extracted, enzymatic hydrolysis, add fructose, peptone, yeast extract, yeast powder, inorganic salts, sweetener etc., after probiotics fermention, adds dietary fiber and prebiotics are made.The probiotics fermention Chinese medicine composition has the effects of inhibiting Growth of Gastric, improving immunity, adjust intestinal flora balance, improve gastrointestinal function, quick, in good taste, good effect.Experiment display, the fermentation composition can dramatically increase the quantity of mouse T lymphocyte, improve immunity of organisms；To stomach cancer cell inhibiting rate up to 65%, illustrate that fermentation composition of the invention is preferable to the therapeutic effect of gastric cancer.

Description

A kind of probiotics fermention Chinese medicine composition and its preparation method and application for treating gastric cancer
Technical field
The present invention relates to a kind of probiotics fermention Chinese medicine compositions and preparation method thereof for treating gastric cancer, belong to traditional Chinese medicine skill Art field.
Background technique
Gastric cancer is one of most common malignant tumour in the world, and in China, its disease incidence occupies the first place of all kinds of tumours, every year about There are 170,000 people to die of gastric cancer, almost close to the 1/4 of whole mortality of malignant tumors numbers, and there are also 20,000 or more new gastric cancers every year Patient generates, and gastric cancer is strictly a kind of disease for seriously threatening people's health.Gastric cancer can betide any age, But common with 40~60 years old, male is more than female, about 2:1.Gastric cancer can betide any position of stomach, but be more common in antrum portion, especially It is lesser curvature side.Untreated person's average life span is about 13 months.
Operation excision is still the only scheme of current radical cure early carcinoma of stomach, and the main means for the treatment of gastric cancer.For a long time with Come, because it is found that gastric cancer is later, most of to belong to late tumor, operative effect is not good enough, and postoperative 5 years survival rates maintain always 30% or so, therefore, it is necessary to seek more more effective therapeutic schemes, to improve survival rate.
The treatment of gastric cancer Chinese medicine is a big characteristic of China's curing gastric cancer, and is ground as China in recent years controls gastric cancer to Chinese medicine That studies carefully deepens continuously so that Chinese medicine treatment plays increasingly important role in the treatment of gastric cancer, Chinese medicine treat gastric cancer from Entirety is set out, and the treatment of gastric cancer Chinese medicine is not only able to be individually used for the treatment of gastric cancer, but also can be with Surgery for Gastric Carcinoma, radiotherapy, chemotherapy Etc. treatment means use in conjunction, can significantly improve comprehensive therapeutic effect, especially mitigate chemicotherapy to the toxic side effect of body, improving Patient immune and the effect for improving its quality of life etc. are obvious, can effectively prevent the Preventive of tumour.
Currently, the related patents of domestic compound of Chinese herbal medicine gastric cancer are existing very much, the Yiqihuoxue Decoction of gastric cancer is such as prevented and treated Chinese materia medica preparation and preparation method thereof-CN201310543123, the Chinese materia medica preparation for treating gastric cancer and preparation method thereof- CN200710052851, a kind of Chinese medicine composition-CN201110262095 treating gastric cancer, a kind of Chinese traditional medicine composition for treating gastric cancer Object-CN201210416744, it a kind of Chinese herbal oral decoction-CN201010003136 for treating gastric cancer, a kind of treats in gastric cancer Drug composition and preparation method thereof-CN201310017007, a kind of Chinese medicine for treating gastric cancer and preparation method thereof- CN201210582510 etc., most of foregoing invention are classical prescription, proved recipe, and production method is simple, extensive, such as decoction, pill, are dissipated Agent, capsule etc., though there is certain curative effect, raw material availability is low, and mouthfeel is very poor, slow effect, brings pole to patient when long-term use Big pain.
Summary of the invention
Gastric cancer category disease of digestive system, patient cause greatly digestive tract environment because of disease itself and various chemicotherapies Damage, has especially upset the ecological balance of intestine beneficial bacteria colony.It lives away from home in normal human's enteron aisle a large amount of microbial population, Wherein probiotics plays a crucial role human health.Probiotics can cannot be decomposed with catabolism upper digestive tract system The food ingredients of metabolism generate short chain fatty acids, are gastrointestinal mucosa cells with nutrient substance.Especially simultaneously for oral drugs It is that certain ingredients of natural drug can carry out catabolism, the precursor substance of drug is transformed into active pharmaceutical ingredient and is sent out The effect of waving.Therefore, if being aided with adjusting intestinal flora while treating disease, undoubtedly great benefit can be brought for patient.
The present invention is to solve the deficiency of existing Chinese medicine prescription, is first carried out to the preferable Chinese prescription of clinical treatment gastric cancer effect High efficiency extraction, using modern biotechnology, by Chinese medicine extract through probiotics fermention, in-vitro simulated human body intestinal canal environment, in advance It to the various effective components in Chinese medicine extract, is decomposed, is directly absorbed after making it into human body, mitigate human body burden, Be equipped with dietary fiber and prebiotics simultaneously, adjust human body intestinal canal environment, develop raw material availability is high, effect is strong, it is in good taste and The high Chinese medicine composition of added value, meets the demand of patient, wide market.
It is an object of the present invention to provide a kind of probiotics fermention Chinese medicine composition for treating gastric cancer, the Chinese medicines Composition is made of the medicinal material of following parts by weight: 3 ~ 6 parts of American Ginseng, 3 ~ 10 parts of radix bupleuri, and 3 ~ 10 parts of Fructus Aurantii, 3 ~ 10 parts of radix scutellariae, half 3 ~ 9 parts of summer, 5 ~ 20 parts of salvia chinensis, 5 ~ 10 parts of sea Piao sheath, 2 ~ 8 parts of fritillaria thunbergii, 6 ~ 15 parts of Radix Paeoniae Alba, 3 ~ 10 parts of Rhizoma Chuanxiong, prepared RHIZOMA CYPERI with vinegar 6 ~ 10 parts, 3 ~ 6 parts of galangal, 6 ~ 12 parts of Radix Angelicae Sinensis, 6 ~ 10 parts of Herba Cistanches, 2 ~ 10 parts of radix glycyrrhizae preparata；Preparation method is as follows:
(1) medicinal material is handled:
A. radix scutellariae, salvia chinensis add the water of 8 ~ 15 times of medicinal material weight together, impregnate 20 ~ 60 minutes, through 115 ~ 121 DEG C of high pressure waters It mentions 30 ~ 60 minutes, filters, filtrate sterilizing is spare for sterile radix scutellariae, salvia chinensis mixed liquor；
B. Radix Angelicae Sinensis adds the water of 8 ~ 15 times of medicinal material weight, and distillate and distillation are collected in immersion 20 ~ distill after sixty minutes respectively Aqueous solution afterwards, distillate sealing, 115 ~ 121 DEG C sterilize 30 ~ 60 minutes, obtain sterile distillate, spare；
C., the Radix Angelicae Sinensis dregs of a decoction are added to the water of 8 ~ 15 times of medicinal material weight together with remaining 12 kinds of medicinal material, are impregnated 20 ~ 60 minutes, warp 115 ~ 121 DEG C of high pressure waters mention 30 ~ 60 minutes, and filtration, the aqueous solution after filtrate is distilled with Radix Angelicae Sinensis merges, and obtain herbal extract；
(2) digest: the herbal extract that step (1) C is obtained maintains 70 ~ 95 DEG C of constant temperature, adjusts pH to 6.0 ~ 7.0, adds Alpha-amylase, enzyme digestion reaction 10 ~ 15 minutes, using iodine solution as indicator, when medical fluid chance iodine solution is constant blue, enzymatic hydrolysis terminated, and adds Heat to 100 DEG C enzyme deactivation 10 ~ 20 minutes, filtering, filtrate is spare；
(3) after the filtrate decompression concentration that step (2) is obtained, fructose, peptone, yeast extract, ferment concentration, ingredient: are added Female powder, add inorganic salts, add sweetener, adjust pH value to 6.5 ~ 7.5,115 ~ 121 DEG C sterilize 30 ~ 60 minutes, obtain sterile medicine Liquid；
(4) it is inoculated with, ferments: when the sterile liquid medicine temperature of step (3) is down to 25 ~ 35 DEG C, (1) B being obtained mutually synthermal Sterile distillate be mixed with, be inoculated with preculture percent by volume be 1% ~ 5% Leuconostoc mesenteroides bacterium solution, 25 ~ 35 DEG C Culture adds glucose when pH value is down to 5.5 ~ 6.5, and high-temperature sterilization is down to 35 ~ 40 DEG C to temperature, is inoculated with the body of preculture A kind of Bifidobacterium bacterium solution that product percentage is 1% ~ 5%, 37 DEG C are cultivated 6 ~ 8 hours, are inoculated with the percent by volume of preculture again For a kind of 1% ~ 5% lactobacillus bacterium solution, continue fermentation 20 ~ 24 hours；When pH value is down to 4.0 ~ 4.1, fermentation temperature is reduced extremely 20 ~ 30 DEG C, continue fermentation to pH value and be down to 3.5 ~ 3.8, terminates fermentation to get Chinese medicine composition fermentation liquid；
(5) it deploys, inactivate: adding sterile dietary fiber, prebiotics and step into the Chinese medicine composition fermentation liquid that (4) obtain Suddenly (1) A is extracted sterile radix scutellariae, salvia chinensis mixed liquor, mix, and 115 ~ 121 DEG C of sterilizings 5 ~ 50 minutes are to get to treatment gastric cancer Probiotics fermention Chinese medicine composition.
Leuconostoc mesenteroides and its tunning can significantly improve animal body GSH-px, SOD activity, reduce serum MDA Level, and the long period is maintained, have the function of improving animal body oxidation resistance, meanwhile, normal intestinal flora can be stimulated Growth, have antagonism to common pathogens such as Shigella, Salmonella, staphylococcus aureuses.
Preferably, the Chinese medicine composition is made of the medicinal material of following parts by weight: 3 parts of American Ginseng, 6 parts of radix bupleuri, and Fructus Aurantii 6 Part, 6 parts of radix scutellariae, 5 parts of the tuber of pinellia, 12 parts of salvia chinensis, 10 parts of sea Piao sheath, 4 parts of fritillaria thunbergii, 9 parts of Radix Paeoniae Alba, 5 parts of Rhizoma Chuanxiong, 6 parts of prepared RHIZOMA CYPERI with vinegar, 4 parts of galangal, 9 parts of Radix Angelicae Sinensis, 10 parts of Herba Cistanches, 2 parts of radix glycyrrhizae preparata；
The amylase is α-alpha-amylase, dosage calculation method are as follows: medicinal material amount × 30/ enzyme-activity unit, before First enzyme is mixed with a small amount of water.
The dosage of the fructose is weight g/ volume ml percentage 0.1 ~ 1.0%, and the dosage of the glucose is weight g/ body Product ml percentage is 0.1 ~ 1.0%.
The dosage of the peptone be weight g/ volume ml percentage 0.05 ~ 1.5%, yeast extract dosage be weight g/ body Accumulate ml percentage 0.01 ~ 0.5%, the dosage of yeast powder is weight g/ volume ml percentage 0.01 ~ 0.5%.
The sweetener is the sucrose and weight g/ volume ml percentage 0.007% of weight g/ volume ml percentage 3% Sucralose.
The inorganic salts are K2HPO43H2O 0.1g/100ml、KH2PO4 0.1g/100ml、FeSO47H2O 5.0mg/ 100ml、ZnSO47H2O 4.4mg/100ml、MgSO47H2O 25mg/100ml。
The dietary fiber is the polydextrose of weight g/ volume ml percentage 1 ~ 5%.
The prebiotics are that the oligofructose of weight g/ volume ml percentage 0.1 ~ 0.7% starches (G 55L).
The Leuconostoc mesenteroides is selected from Leuconostoc mesenteroides dextran subspecies or Leuconostoc mesenteroides butterfat is sub- Kind；The Bifidobacterium is selected from bifidobacterium bifidum, bifidobacterium breve, bifidobacterium adolescentis or bifidobacterium infantis；It is described Lactobacillus be selected from lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus casei, Lactobacillus rhamnosus, lactobacillus reuteri or plant Lactobacillus.
It is also another object of the present invention to provide the probiotics fermention Chinese medicine compositions of the treatment gastric cancer to press down in preparation Growth of Gastric processed improves immunity, adjusts intestinal flora balance, improving the application in the drug of gastrointestinal function.It can be according to Known conventional method, by the probiotics fermention Chinese medicine composition of the treatment gastric cancer, add it is one or more have it is identical, Similar or different biological is active, and be presented with auxiliary or synergistic effect to primary activity ingredient, but mutual not antagonism again each other Or mixtures thereof natural or synthetic other function composition is made and is suitable for oral mixture, emulsion, pill, powder, capsule, piece Agent or granule.
The invention has the following advantages that
1, the probiotics fermention Chinese medicine composition has quick, in good taste, inhibition Growth of Gastric, raising immune The effects of power adjusts intestinal flora balance, improves gastrointestinal function.
2, it when the present invention is to traditional Chinese medicine extyaction, according to the characteristic of medicinal material, is extracted, is mentioned using different technique The high utilization rate of medicinal material.
3, present invention simulation human body intestinal canal environment, is handled Chinese medicine extract first with zymotechnic, using it as training Base is supported, batch fermentation is carried out using more probiotics, becomes feasible to the effective component of Chinese medicine by the active matter of intestinal absorption Matter, quick, good effect reduce human body intestinal canal burden, compensate for the deficiency of existing Chinese medicine product slow effect, period length.
4, the water-soluble dietary fiber polydextrose and prebiotic, fructooligosaccharide not being absorbed by the body are added in this composition, Sweet mouthfeel is agreeable to the taste, in stable condition, can significantly improve human body intestinal canal environment, improves the quality of life of patient.
5, this composition is not added with any preservative, and through high-temperature sterilization, sterile filling, long shelf-life is drunk conveniently.
Specific embodiment
The following describes the present invention further through the description of specific embodiments, but it is to limit of the invention that this, which is not, System, those skilled in the art's basic thought according to the present invention can make various modifications or improvements, but without departing from this The basic thought of invention, is all within the scope of the present invention；Material described in embodiment meets respective country standard.
Embodiment 1: the preparation of the probiotics fermention Chinese medicine composition A of gastric cancer is treated
Be made of the medicinal material of following parts by weight: 3 parts of American Ginseng, 4 parts of radix bupleuri, 4 parts of Fructus Aurantii, 4 parts of radix scutellariae, 4 parts of the tuber of pinellia, stone is shown in Wear 8 parts, 10 parts of sea Piao sheath, 4 parts of fritillaria thunbergii, 8 parts of Radix Paeoniae Alba, 4 parts of Rhizoma Chuanxiong, 4 parts of prepared RHIZOMA CYPERI with vinegar, 3 parts of galangal, 7 parts of Radix Angelicae Sinensis, Herba Cistanches 7 parts, 2 parts of radix glycyrrhizae preparata, it is specific the preparation method is as follows:
(1) medicinal material is handled: radix scutellariae, salvia chinensis add the water of 8 times of medicinal material amounts together, impregnate 30 minutes, mention through 115 DEG C of high pressure waters 60 minutes, filtration, it was sterile radix scutellariae, salvia chinensis mixed liquor that 115 DEG C of filtrate sterilizings sterilizing in 60 minutes is spare；Radix Angelicae Sinensis adds 8 times of medicinal materials The water of amount distills after impregnating 30 minutes, and the aqueous solution after collecting distillate and distillation respectively is spare, and distillate sealing, 115 DEG C go out Bacterium 60 minutes, obtain sterile distillate；The water that the Radix Angelicae Sinensis dregs of a decoction are added to 8 times of medicinal material amounts together with remaining 12 kinds of medicinal materials, impregnates 30 minutes, It mentions 60 minutes, filters through 115 DEG C of high pressure waters, the aqueous solution after filtrate is distilled with Radix Angelicae Sinensis merges with high pressure aqueous extract, obtains medicinal material Extracting solution.
(2) it digests: by (1) Chinese medicine extracting solution, maintaining 85 DEG C of constant temperature, adjust pH to 6.5, increase temperature amylase, digest Reaction 10 ~ 15 minutes, using iodine solution as indicator, when medical fluid chance iodine solution is constant blue, enzymatic hydrolysis terminates, and is heated to 100 DEG C of enzyme deactivations 10 Minute, it filters, filtrate is spare.
(3) concentration, ingredient: will in (2) filtrate decompression concentration after, addition fructose 0.6%(weight g/ volume ml), peptone 0.3%(weight g/ volume ml), yeast extract 0.1%(weight g/ volume ml), yeast powder 0.06%(weight g/ volume ml), inorganic salts (K2HPO43H2O 0.1g/100ml、KH2PO4 0.1g/100ml、FeSO47H2O 5.0mg/100ml、ZnSO47H2O 4.4mg/ 100ml、MgSO47H2O 25mg/100ml), sucrose 3.0%(weight g/ volume ml) and Sucralose 0.007%(weight g/ volume Ml).Adjust pH value to 7.0,115 DEG C sterilize 60 minutes, obtain sterile liquid medicine.
(4) it is inoculated with, ferments: when sterile liquid medicine temperature is down to 30 DEG C, (1) middle mutually synthermal sterile distillate being mixed therewith Close, be inoculated with preculture percent by volume be 2% Leuconostoc mesenteroides dextran subspecies (CICC 22179) bacterium solution, 30 DEG C culture, when pH value is down to 5.5, add glucose 0.6%(weight g/ volume ml), 115 DEG C sterilize 30 minutes, to temperature drop To 38 DEG C, it is inoculated with bifidobacterium breve (AS 1.2213) bacterium solution that the percent by volume of preculture is 1%, 37 DEG C are cultivated 6 hours, then Lactobacillus plantarum (AS 1.19) bacterium solution that the percent by volume of secondary inoculation preculture is 1%, continues fermentation 20 hours.When pH value drops When to 4.0, fermentation temperature is reduced to 25 DEG C, continues fermentation to pH value and is down to 3.8, terminates fermentation to get Chinese medicine composition fermentation Liquid.
(5) deploy, inactivate: the Chinese medicine composition fermentation liquid in (4) adds the polydextrose 4%(weight g/ to sterilize in advance Volume ml), oligofructose starch (G 55L) 0.6%(weight g/ volume ml) and (1) in extract sterile radix scutellariae, salvia chinensis mixing Liquid mixes, and 115 DEG C sterilize 20 minutes, must treat the probiotics fermention Chinese medicine composition of gastric cancer, (rule filling under aseptic condition Lattice are 100ml/ bottles).
Embodiment 2: the preparation of the probiotics fermention Chinese medicine composition B of gastric cancer is treated
Be made of the medicinal material of following parts by weight: 3 parts of American Ginseng, 7 parts of radix bupleuri, 7 parts of Fructus Aurantii, 4 parts of radix scutellariae, 6 parts of the tuber of pinellia, stone is shown in Wear 15 parts, 12 parts of sea Piao sheath, 4 parts of fritillaria thunbergii, 9 parts of Radix Paeoniae Alba, 5 parts of Rhizoma Chuanxiong, 6 parts of prepared RHIZOMA CYPERI with vinegar, 5 parts of galangal, 6 parts of Radix Angelicae Sinensis, meat desert 8 parts of Rong, 2 parts of radix glycyrrhizae preparata, it is specific the preparation method is as follows:
(1) medicinal material is handled: radix scutellariae, salvia chinensis add the water of 10 times of medicinal material amounts together, impregnate 60 minutes, through 115 DEG C of high pressure waters It mentions 50 minutes, filters, it is sterile radix scutellariae, salvia chinensis mixed liquor that 121 DEG C of filtrate sterilizings sterilizing in 30 minutes is spare；Radix Angelicae Sinensis adds 10 times The water of medicinal material amount, immersion are distilled after sixty minutes, and the aqueous solution after collecting distillate and distillation respectively is spare, distillate sealing, and 121 DEG C sterilizing 30 minutes, obtain sterile distillate；The water that the Radix Angelicae Sinensis dregs of a decoction are added to 10 times of medicinal material amounts together with remaining 12 kinds of medicinal materials, impregnates 60 Minute, it mentions 50 minutes, filters through 115 DEG C of high pressure waters, the aqueous solution after filtrate is distilled with Radix Angelicae Sinensis merges, and obtains herbal extract.
(2) it digests: by (1) Chinese medicine extracting solution, maintaining 85 DEG C of constant temperature, adjust pH to 6.0, increase temperature amylase, digest Reaction 10 ~ 15 minutes, using iodine solution as indicator, when medical fluid chance iodine solution is constant blue, enzymatic hydrolysis terminates, and is heated to 100 DEG C of enzyme deactivations 15 Minute, it filters, filtrate is spare.
(3) concentration, ingredient: will in (2) filtrate decompression concentration after, addition fructose 0.5%(weight g/ volume ml), peptone 0.2%(weight g/ volume ml), yeast extract 0.1%(weight g/ volume ml), yeast powder 0.06%(weight g/ volume ml), inorganic salts (K2HPO43H2O 0.1g/100ml、KH2PO4 0.1g/100ml、FeSO47H2O 5.0mg/100ml、ZnSO47H2O 4.4mg/ 100ml、MgSO47H2O 25mg/100ml), sucrose 3.0%(weight g/ volume ml) and Sucralose 0.007%(weight g/ volume Ml).Adjust pH value to 7.2,121 DEG C sterilize 30 minutes, obtain sterile liquid medicine.
(4) it is inoculated with, ferments: when sterile liquid medicine temperature is down to 28 DEG C, (1) middle mutually synthermal sterile distillate being mixed therewith Close, be inoculated with preculture percent by volume be 4% Leuconostoc mesenteroides dextran subspecies (CICC 22179) bacterium solution, 28 DEG C culture, when pH value is down to 5.5, add glucose, 115 DEG C sterilize 30 minutes, be down to 37 DEG C to temperature, be inoculated with preculture Bifidobacterium bifidum (normal human's separation, identified through Institute of Microorganism, Academia Sinica) bacterium solution that percent by volume is 1%, 37 DEG C are cultivated 8 hours, are inoculated with Lactobacillus delbrueckii (AS 1.1480) bacterium solution that the percent by volume of preculture is 1% again, are continued Fermentation 24 hours.When pH value is down to 4.0, fermentation temperature is reduced to 28 DEG C, is continued fermentation to pH value and is down to 3.7, terminates fermentation, Up to Chinese medicine composition fermentation liquid.
(5) deploy, inactivate: the Chinese medicine composition fermentation liquid in (4) adds the polydextrose 5%(weight g/ to sterilize in advance Volume ml), oligofructose starch (G 55L) 0.5%(weight g/ volume ml) and (1) in extract sterile radix scutellariae, salvia chinensis mixing Liquid mixes, and 115 DEG C sterilize 15 minutes, must treat the probiotics fermention Chinese medicine composition of gastric cancer, (rule filling under aseptic condition Lattice are 100ml/ bottles).
Embodiment 3: the preparation of the probiotics fermention Chinese medicine composition C of gastric cancer is treated
Be made of the medicinal material of following parts by weight: 3 parts of American Ginseng, 6 parts of radix bupleuri, 6 parts of Fructus Aurantii, 6 parts of radix scutellariae, 5 parts of the tuber of pinellia, stone is shown in Wear 12 parts, 10 parts of sea Piao sheath, 4 parts of fritillaria thunbergii, 9 parts of Radix Paeoniae Alba, 5 parts of Rhizoma Chuanxiong, 6 parts of prepared RHIZOMA CYPERI with vinegar, 4 parts of galangal, 9 parts of Radix Angelicae Sinensis, meat desert 10 parts of Rong, 2 parts of radix glycyrrhizae preparata, it is specific the preparation method is as follows:
(1) medicinal material is handled: radix scutellariae, salvia chinensis add the water of 12 times of medicinal material amounts together, impregnate 60 minutes, through 115 DEG C of high pressure waters Mention 60 minutes, filter, 115 DEG C of filtrate sterilizing 50 minutes it is spare, be sterile radix scutellariae, salvia chinensis mixed liquor；Radix Angelicae Sinensis adds 12 times of medicinal materials The water of amount, immersion are distilled after sixty minutes, and the aqueous solution after collecting distillate and distillation respectively is spare, and distillate sealing, 115 DEG C go out Bacterium 50 minutes, obtain sterile distillate；The water that the Radix Angelicae Sinensis dregs of a decoction are added to 12 times of medicinal material amounts together with remaining 12 kinds of medicinal materials, impregnates 60 points Clock mentions 60 minutes through 115 DEG C of high pressure waters, and filtration, the aqueous solution after filtrate is distilled with Radix Angelicae Sinensis merges, and obtains herbal extract.
(2) it digests: by (1) Chinese medicine extracting solution, maintaining 85 DEG C of constant temperature, adjust pH to 6.0, increase temperature amylase, digest Reaction 10 ~ 15 minutes, using iodine solution as indicator, when medical fluid chance iodine solution is constant blue, enzymatic hydrolysis terminates, and is heated to 100 DEG C of enzyme deactivations 10 Minute, it filters, filtrate is spare.
(3) concentration, ingredient: will in (2) filtrate decompression concentration after, addition fructose 0.4%(weight g/ volume ml), peptone 0.3%(weight g/ volume ml), yeast extract 0.08%(weight g/ volume ml), yeast powder 0.04%(weight g/ volume ml), inorganic salts (K2HPO43H2O 0.1g/100ml、KH2PO4 0.1g/100ml、FeSO47H2O 5.0mg/100ml、ZnSO47H2O 4.4mg/ 100ml、MgSO47H2O 25mg/100ml), sucrose 3.0%(weight g/ volume ml) and Sucralose 0.007%(weight g/ volume Ml).Adjust pH value to 7.5,115 DEG C sterilize 50 minutes, obtain sterile liquid medicine.
(4) it is inoculated with, ferments: when sterile liquid medicine temperature is down to 30 DEG C, (1) middle mutually synthermal sterile distillate being mixed therewith It closes, is inoculated with leuconostoc mesenteroides sub species cremoris (CICC 22181) bacterium solution that the percent by volume of preculture is 3%, 30 DEG C are cultivated, When pH value is down to 5.8, glucose 0.4%(weight g/ volume ml is added), 115 DEG C sterilize 30 minutes, 37 DEG C are down to temperature, It is inoculated with bifidobacterium adolescentis (AS 1.2190) bacterium solution that the percent by volume of preculture is 1%, 37 DEG C are cultivated 7 hours, are connect again Lactobacillus acidophilus (AS 1.1854) bacterium solution that the percent by volume of kind preculture is 1%, continues fermentation 22 hours.When pH value is down to When 4.1, fermentation temperature is reduced to 25 DEG C, continues fermentation to pH value and is down to 3.8, terminates fermentation to get Chinese medicine composition fermentation liquid.
(5) deploy, inactivate: the Chinese medicine composition fermentation liquid in (4) adds the polydextrose 4%(weight g/ to sterilize in advance Volume ml), oligofructose starch (G 55L) 0.5%(weight g/ volume ml) and (1) in extract sterile radix scutellariae, salvia chinensis mixing Liquid mixes, and 115 DEG C sterilize 15 minutes, must treat the probiotics fermention Chinese medicine composition of gastric cancer, (rule filling under aseptic condition Lattice are 100ml/ bottles).
Experimental example 1: fermented tcm composition of the invention tests the function and effect of immunity of organisms
This experiment is measured mainly for function and effect of the object of the present invention to immunity of organisms；Control sample be with it is excellent Choose the unleavened high pressure extraction object of medicine component solutions same concentrations；It is commercially available for testing drug and instrument used.
The preparation of 1.1 mice spleen suspensions
Balb/c mouse is put to death, is impregnated 5 minutes with 75% alcohol, the sterile complete spleen of separation washes away floating blood with PBS, shells Except excess tissue ingredient, 5mlPBS is inhaled with syringe, is inserted into spleen, cell is blown out, is repeated several times, remaining spleen transparent to spleen outer membrane Tissue is cut into size 1mm3Fritter shreds and is placed on 200 mesh filter screens, is ground with syringe needle core, and cleaning solution is collected in PBS washing, 1200rpm is centrifuged 5 minutes, abandons supernatant, adds 3ml erythrocyte cracked liquid, stands 2 minutes, and 10mlPBS is added to terminate reaction, 1200rpm Supernatant is abandoned in centrifugation 5 minutes, and PBS is washed twice, and 1200rpm is centrifuged 5 minutes, is washed once with RPMI-1640 incomplete culture medium, 1200rpm is centrifuged 5 minutes, abandons supernatant, adds appropriate RPMI-1640 complete medium (containing 10% fetal calf serum, 100u/ml mould Element, 100ug/ml streptomysin and 10mM Hepes), adjustment cell concentration to 2 × 104A cell/ml, Trypan Blue, detection Cell survival rate is greater than 95%.
1.2 fermented tcm compositions of the present invention are on lymphopoietic influence
Fermented tcm composition A, B, C and control group prepared by embodiment 1 ~ 3 are subjected to gradient dilution with pure water respectively, obtained Respective gradient dilution liquid.Above-mentioned splenocyte suspension is added in 96 well culture plates by the hole 100ul/, then is added respectively 10ulConA(5ug/ml), the gradient dilution liquid of the 10ul present composition, each gradient do 6 repetitions, culture plate are placed in 37℃、5%CO2Incubator in cultivate 72h；Blank control (addition 10ul pure water) is set simultaneously.Using the kit side CCK-8 Method measures influence (with stimulus index SI indicate) of each fermented product to spleen lymphocyte proliferation ability, and the results are shown in Table 1.
Table 1 treats the probiotics fermention Chinese medicine composition of gastric cancer to lymphopoietic influence
Stimulus index SI value and splenic lymphocytes how much being positively correlated property, as shown in Table 1, with the unfermentable Chinese medicine of control group Extracting solution is compared, and the SI value of fermented tcm composition of the invention is all larger than 4, hence it is evident that is higher than control group, is illustrated that it can be significant Increase the quantity of mouse T lymphocyte, improves immunity of organisms.
Experimental example 2: fermented tcm composition of the invention tests Growth of Gastric inhibitory effect
This experiment is measured Growth of Gastric inhibitory effect mainly for fermented tcm composition of the invention；It is right It is and the unfermentable high pressure extraction object of preferred traditional Chinese medicinal components scheme same concentrations according to group sample；Drug concentration, which is pressed, contains dry medicinal material meter It calculates, animal dosage with 10 times of drug ports of clinical equivalent dosage quite to raise animal, and dosage is big or hour progress is appropriate dense Contracting or dilution；It is commercially available for testing drug and instrument used.
2.1 the preparation of drug containing animal blood serum
Rat 196, male, weight 250 ~ 300g/ only, is randomly divided into 14 groups, and every group 14, gastric infusion；(1) blank Control group fills the physiological saline with experimental group same dose；(2) experimental group is fermentation group (in fermented tcm composition A, fermentation Drug composition B, fermented tcm composition C), group of not fermenting (drug control group) (basic, normal, high concentration is respectively 3.235, 6.470,12.94g/kg7d), 2ml/ times/day of dosage, continuous 7 days, administration took blood after 6 hours every time, two repeated every time； (3) 5 FU 5 fluorouracil (5-Fu) positive controls, 50mg/kg7d, intraperitoneal injection；Broken end takes blood, in 4 DEG C, 2000r/min Be centrifuged 20min, separate serum, dispense after 56 DEG C of inactivation 30min, -20 DEG C freeze it is spare.
2.2 cell inhibitory effects test (MTT reduction method)
The SGC-7901 cell of logarithmic growth phase, it is outstanding with the RPMI-1640 culture solution adjustment cell containing 10% fetal calf serum Liquid concentration is 2.0 × 104A/L inoculating cell sets 37 DEG C, 5% CO in 96 well culture plates, every hole 200ul2, the training of saturated humidity It supports in case after culture for 24 hours, replacement is further cultured for for 24 hours, making cell synchronization without the 1640 culture medium of fetal calf serum；Supernatant is abandoned, respectively Group is changed respectively to contain the RPMI-1640 culture solution of 10% corresponding rat blood serum, and every group sets 6 multiple holes culture 48h；Every hole is added again 5mg/ml MTT solution 20ul continues to be incubated for 4h, adds 200ul DMSO after abandoning supernatant, and micro oscillator vibrates 10min, stands 490nm wavelength absorbance value (OD) is measured with microplate reader, calculates cell inhibitory rate.
Cell inhibitory rate (IR)=1-(experimental group mean absorbance values/blank control group mean absorbance values) } × 100%.
The influence result that 2.3 fermented tcm compositions of the present invention are proliferated Human gastric cancer SGC-7901 cells strain:
As shown in Table 2, positive controls (5-Fu) are 66% to the inhibiting rate of stomach cancer cell, and fermentation group (height) is thin to gastric cancer Born of the same parents' inhibiting rate is about 65%, and inhibiting rate is not much different compared with existing chemicals (5-Fu), and group of not fermenting (height) is to stomach cancer cell Inhibiting rate is about 51%, and fermentation group is higher than group of not fermenting, illustrates that preferably (prepared by embodiment 3 for therapeutic effect of this composition to gastric cancer Fermentation composition C effect it is best)；Positive controls (5-Fu) reach the inhibition of stomach cancer cell when 5 days serum is administered Highest and later maintenance balance, fermentation group inhibit to reach highest when 4 days serum is administered and maintain to put down later to stomach cancer cell Weighing apparatus, group of not fermenting inhibit unstable always to stomach cancer cell, illustrate that fermentation composition of the present invention does not ferment a group phase with chemicals, It is rapid, more quick than absorbing.
The inhibiting rate (%) that 2 present composition of table is proliferated Human gastric cancer SGC-7901 cells strain
Note: the time (day) is that number of days is administered in rat in table.

Claims (4)

1. a kind of probiotics fermention Chinese medicine composition for treating gastric cancer, which is characterized in that the Chinese medicine composition by weighing as follows The medicinal material of amount part is made: 3 parts of American Ginseng, 6 parts of radix bupleuri, and 6 parts of Fructus Aurantii, 6 parts of radix scutellariae, 5 parts of the tuber of pinellia, 12 parts of salvia chinensis, sea Piao sheath 10 Part, 4 parts of fritillaria thunbergii, 9 parts of Radix Paeoniae Alba, 5 parts of Rhizoma Chuanxiong, 6 parts of prepared RHIZOMA CYPERI with vinegar, 4 parts of galangal, 9 parts of Radix Angelicae Sinensis, 10 parts of Herba Cistanches, radix glycyrrhizae preparata 2 Part；Preparation method is as follows:
(1) medicinal material is handled:
A. radix scutellariae, salvia chinensis add the water of 8 ~ 15 times of medicinal material weight together, impregnate 20 ~ 60 minutes, mention 30 through 115 ~ 121 DEG C of high pressure waters ~ 60 minutes, filtration, filtrate sterilized, spare for sterile radix scutellariae, salvia chinensis mixed liquor；
B. Radix Angelicae Sinensis adds the water of 8 ~ 15 times of medicinal material weight, immersion 20 ~ distill after sixty minutes, the water after collecting distillate and distillation respectively Solution, distillate sealing, 115 ~ 121 DEG C sterilize 30 ~ 60 minutes, obtain sterile distillate, spare；
C., the Radix Angelicae Sinensis dregs of a decoction add to the water of 8 ~ 15 times of medicinal material weight together with remaining 12 kinds of medicinal material, impregnate 20 ~ 60 minutes, through 115 ~ 121 DEG C of high pressure waters mention 30 ~ 60 minutes, and filtration, the aqueous solution after filtrate is distilled with Radix Angelicae Sinensis merges, and obtain herbal extract；
(2) digest: the herbal extract that step (1) C is obtained maintains 70 ~ 95 DEG C of constant temperature, adjusts pH to 6.0 ~ 7.0, increases temperature Amylase, enzyme digestion reaction 10 ~ 15 minutes, using iodine solution as indicator, when medical fluid chance iodine solution is constant blue, enzymatic hydrolysis terminated, and is heated to 100 DEG C enzyme deactivation 10 ~ 20 minutes, filtering, filtrate is spare；
(3) concentration, ingredient: after the filtrate decompression concentration that step (2) is obtained, adding fructose, peptone, yeast extract, yeast powder, Add inorganic salts, add sweetener, adjust pH value to 6.5 ~ 7.5,115 ~ 121 DEG C sterilize 30 ~ 60 minutes, obtain sterile liquid medicine；
(4) it is inoculated with, ferments: when the sterile liquid medicine temperature of step (3) is down to 25 ~ 35 DEG C, the mutually synthermal nothing that (1) B is obtained Bacterium distillate is mixed with, and is inoculated with the Leuconostoc mesenteroides bacterium solution that the percent by volume of preculture is 1% ~ 5%, and 25 ~ 35 DEG C are cultivated, When pH value is down to 5.5 ~ 6.5, glucose is added, high-temperature sterilization is down to 35 ~ 40 DEG C to temperature, is inoculated with the volume hundred of preculture Point compare a kind of Bifidobacterium bacterium solution for 1% ~ 5%, 37 DEG C are cultivated 6 ~ 8 hours, again the percent by volume of inoculation preculture for 1% ~ A kind of 5% lactobacillus bacterium solution, continues fermentation 20 ~ 24 hours；When pH value is down to 4.0 ~ 4.1, fermentation temperature is reduced to 20 ~ 30 DEG C, continue fermentation to pH value and be down to 3.5 ~ 3.8, terminates fermentation to get Chinese medicine composition fermentation liquid；
(5) it deploys, inactivate: adding sterile dietary fiber, prebiotics and step into the Chinese medicine composition fermentation liquid that (4) obtain (1) A extract sterile radix scutellariae, salvia chinensis mixed liquor, mix, 115 ~ 121 DEG C sterilizing 5 ~ 50 minutes to get；
In above-mentioned steps, the dosage of the fructose is weight g/ volume ml percentage 0.1 ~ 1.0%, and the dosage of the glucose is Weight g/ volume ml percentage is 0.1 ~ 1.0%；
In above-mentioned steps, the dosage of the peptone is weight g/ volume ml percentage 0.05 ~ 1.5%, the dosage of yeast extract is attached most importance to Measure g/ volume ml percentage 0.01 ~ 0.5%, the dosage of yeast powder is weight g/ volume ml percentage 0.01 ~ 0.5%；The sweet tea Taste agent is the sucrose of weight g/ volume ml percentage 3% and the Sucralose of weight g/ volume ml percentage 0.007%；
In above-mentioned steps, the inorganic salts are K2HPO4•3H2O 0.1g/100ml、KH2PO4 0.1g/100ml、FeSO4•7H2O 5.0mg/100ml、ZnSO4•7H2O 4.4mg/100ml、MgSO4•7H2O 25mg/100ml。
2. the probiotics fermention Chinese medicine composition for the treatment of gastric cancer according to claim 1, which is characterized in that the goldbeater's skin Leukonid is selected from Leuconostoc mesenteroides dextran subspecies or leuconostoc mesenteroides sub species cremoris；The Bifidobacterium choosing From bifidobacterium bifidum, bifidobacterium breve, bifidobacterium adolescentis or bifidobacterium infantis；The lactobacillus is selected from acidophilus cream bar Bacterium, Lactobacillus delbrueckii, Lactobacillus casei, Lactobacillus rhamnosus, lactobacillus reuteri or lactobacillus plantarum.
3. a kind of preparation method of probiotics fermention Chinese medicine composition for treating gastric cancer of any of claims 1 or 2, feature exist In steps are as follows:
(1) medicinal material is handled:
A. radix scutellariae, salvia chinensis add the water of 8 ~ 15 times of medicinal material weight together, impregnate 20 ~ 60 minutes, mention 30 through 115 ~ 121 DEG C of high pressure waters ~ 60 minutes, filtration, filtrate sterilized, spare for sterile radix scutellariae, salvia chinensis mixed liquor；
B. Radix Angelicae Sinensis adds the water of 8 ~ 15 times of medicinal material weight, immersion 20 ~ distill after sixty minutes, the water after collecting distillate and distillation respectively Solution, distillate sealing, 115 ~ 121 DEG C sterilize 30 ~ 60 minutes, obtain sterile distillate, spare；
C., the Radix Angelicae Sinensis dregs of a decoction add to the water of 8 ~ 15 times of medicinal material weight together with remaining 12 kinds of medicinal material, impregnate 20 ~ 60 minutes, through 115 ~ 121 DEG C of high pressure waters mention 30 ~ 60 minutes, and filtration, the aqueous solution after filtrate is distilled with Radix Angelicae Sinensis merges, and obtain herbal extract；
(2) digest: the herbal extract that step (1) C is obtained maintains 70 ~ 95 DEG C of constant temperature, adjusts pH to 6.0 ~ 7.0, increases temperature Amylase, enzyme digestion reaction 10 ~ 15 minutes, using iodine solution as indicator, when medical fluid chance iodine solution is constant blue, enzymatic hydrolysis terminated, and is heated to 100 DEG C enzyme deactivation 10 ~ 20 minutes, filtering, filtrate is spare；
(3) concentration, ingredient: after the filtrate decompression concentration that step (2) is obtained, adding fructose, peptone, yeast extract, yeast powder, Add inorganic salts, add sweetener, adjust pH value to 6.5 ~ 7.5,115 ~ 121 DEG C sterilize 30 ~ 60 minutes, obtain sterile liquid medicine；
(4) it is inoculated with, ferments: when the sterile liquid medicine temperature of step (3) is down to 25 ~ 35 DEG C, the mutually synthermal nothing that (1) B is obtained Bacterium distillate is mixed with, and is inoculated with the Leuconostoc mesenteroides bacterium solution that the percent by volume of preculture is 1% ~ 5%, and 25 ~ 35 DEG C are cultivated, When pH value is down to 5.5 ~ 6.5, glucose is added, high-temperature sterilization is down to 35 ~ 40 DEG C to temperature, is inoculated with the volume hundred of preculture Point compare a kind of Bifidobacterium bacterium solution for 1% ~ 5%, 37 DEG C are cultivated 6 ~ 8 hours, again the percent by volume of inoculation preculture for 1% ~ A kind of 5% lactobacillus bacterium solution, continues fermentation 20 ~ 24 hours；When pH value is down to 4.0 ~ 4.1, fermentation temperature is reduced to 20 ~ 30 DEG C, continue fermentation to pH value and be down to 3.5 ~ 3.8, terminates fermentation to get Chinese medicine composition fermentation liquid；
(5) it deploys, inactivate: adding sterile dietary fiber, prebiotics and step into the Chinese medicine composition fermentation liquid that (4) obtain (1) A extract sterile radix scutellariae, salvia chinensis mixed liquor, mix, 115 ~ 121 DEG C sterilizing 5 ~ 50 minutes to get.
4. it is of any of claims 1 or 2 treatment gastric cancer probiotics fermention Chinese medicine composition preparation inhibit Growth of Gastric, Improve the application in the drug of immunity.
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