CN103260632B - Gastrin formation inhibitor and the food compositions containing this gastrin formation inhibitor - Google Patents

Gastrin formation inhibitor and the food compositions containing this gastrin formation inhibitor Download PDF

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CN103260632B
CN103260632B CN201280003716.6A CN201280003716A CN103260632B CN 103260632 B CN103260632 B CN 103260632B CN 201280003716 A CN201280003716 A CN 201280003716A CN 103260632 B CN103260632 B CN 103260632B
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gastrin
formation inhibitor
food
bacterium
present
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CN103260632A (en
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古贺泰裕
浅见幸夫
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Meiji Co Ltd
Tokai University Educational System
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Meiji Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065—Microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/145—Gasseri

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  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
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Abstract

Problem of the present invention is, provides a kind of novel medicine or food, and it does not utilize the anti-Hp effect of milk-acid bacteria, and prevention improves the series of symptoms such as pyrosis, stomachache, reflux esophagitis, the esophageal carcinoma etc. that hyperchlorhydria causes.The present invention relates to the gastrin formation inhibitor of the dead thalline comprising genus lactubacillus bacterium and the food compositions containing this gastrin formation inhibitor.Especially, the present invention relates to the gastrin formation inhibitor that genus lactubacillus bacterium is Lactobacillus gasseri (especially Lactobacillus gasseri OLL2716).

Description

Gastrin formation inhibitor and the food compositions containing this gastrin formation inhibitor
Technical field
The present invention relates to the milk-acid bacteria with the effect improving gastritis, stomach ulcer, hyperchlorhydria and reflux esophagitis, the pharmaceutical composition comprising this milk-acid bacteria and food compositions.
Background technology
Comprise in enteron aisle and be rich in multifarious microorganism species, described flora is formed by the multiple different strains comprising the bacterium that high-density is survived, and the concentration of this flora reaches 10 of tube chamber content 11~ 10 12cells/g.Fully know, have an instinct for the microorganism species possessed in digestive tube and effect that is healthy, disease is comprised: nourish metabolic activity, gut epithelium and immune effect and protect the host of its Su Ju not to be subject to (non-patent literatures 1) such as the intrusions of external microorganism.
On the other hand, when stomach does not infect helicobacter pylori, in human body, only there are several bacteriums.Go on a hunger strike period, gastric juice only comprise comprise Streptococcus, Lactobacillus and Veillonella, about 10 2~ 10 3the minority bacterium of/ml.But these bacteriums are the momentary bacteriums being derived from oral cavity, throat, uncommon (non-patent literature 2 and 3).Can think people stomach in lack this bacterium be the reason causing acidity in tube chamber high.
Fully know, helicobacter pylori is the pathogenic bacteria causing peptide ulceration, cancer in the stomach of people.It is also proposed Helicobacter bacteria to belong to people all the time and have an instinct for microorganism species (non-patent literature 4) in the stomach that possesses.This hypothesis is subject to following true support: helicobacter pylori just obtains at the early stage of infantile period and continue Su Ju under one's belt with suitable quantity within decades subsequently.Thus, for physiological development and the function of stomach, need to understand fully that stomach has an instinct for the effect of the microorganism species possessed.But the infection experiment be difficult to by employing helicobacter pylori understands fully that it acts on.This is because the various pathogenic agents of the helicobacter pylori of CagA, vacuolating cytotoxin, urease and meta-bolites thereof etc. bring out the inflammation of chronic disease in stomach-tissue, and this situation makes helicobacter pylori thicken as having an instinct for the bacterial physiology effect possessed.
In previous research, the present inventor etc. find in the stomach of specific-pathogen free (specificpathogen-free, SPF) mouse dominant to be formed by genus lactubacillus bacterium have an instinct for the microorganism species (non-patent literature 5) possessed.Can think, because the acidity of the stomach of SPF mouse is lower, therefore genus lactubacillus bacterium can Su Ju under one's belt.And then, create agnogenic inflammatory activity in the stomach of SPF mouse.
Like this, comprise the milk-acid bacteria of multiple research groups for genus lactubacillus bacterium etc. of the present inventor etc., Hp (helicobacter pylori) and the relation of stomach, be repeated research.Wherein, large quantifier elimination (patent documentation 1 ~ 5) has been carried out in the anti-improvement of giving for the gastritis caused by milk-acid bacteria, stomach ulcer.But the anti-Hp effect of milk-acid bacteria of these a large amount of research and utilizations, give anti-improvement for what do not utilize the disease of upper digestive tract of described effect etc., study not yet fully, this is present situation.
Prior art document
Patent documentation
Patent documentation 1: Japanese Patent No. 4509250 publication
Patent documentation 2: Japanese Patent No. 3046303 publication
Patent documentation 3: Japanese Patent No. 2672247 publication
Patent documentation 4: Japanese Patent No. 3810403 publication
Patent documentation 5: Japanese Patent No. 4021951 publication
Non-patent literature
Non-patent literature 1:GuarnerF, MalageladaJR. " Gutflorainhealthanddisease. " Lancet2003; 361:512-519.
Non-patent literature 2:LambertJ, andHullR. " Uppergastrointestinaltractdiseaseandprobiotics " AsiaPacificJClinNutr1996; 5:31-35.
Non-patent literature 3:RuddellWS, AxonAT, FindlayJM, BartholomewBA, andHillMJ. " Effectofcimetidineonthegastricbacterialflora. " Lancet.1980; Mar29:672-674.
Non-patent literature 4:BlaserMJ. " Helicobactersareindigenoustothehumanstomach:duodenalulce rationisduetochangesingastricmicroecologyinthemodernera. " Gut1998; 43:721-727.
Non-patent literature 5:KabirAMA, AibaY, TakagiA, KamiyaS, MiwaT, andKogaY. " PreventionofHelicobacterpyloriinfectionbylactobacilliina gnotobioticmurinemodel. " Gut1997; 41:49-55.
Summary of the invention
the problem that invention will solve
The discoveries such as the present inventor: when giving genus lactubacillus bacterium with the form of viable bacteria or dead bacterium to axenic mice, the cell count of the gastrin generated in stomach significantly reduces, and then repeats further investigation, and its result, completes the present invention.
for the scheme of dealing with problems
That is, the present invention relates to following gastrin formation inhibitor and the food compositions containing this gastrin formation inhibitor.
[1] a gastrin formation inhibitor, it comprises the dead thalline of genus lactubacillus bacterium.
[2] a gastrin formation inhibitor, its comprise genus lactubacillus bacterium dead thalline, do not comprise viable bacteria body.
[3] the gastrin formation inhibitor according to aforementioned [1] or [2], wherein, genus lactubacillus bacterium is Lactobacillus gasseri.
[4] the gastrin formation inhibitor according to aforementioned [1] or [2], wherein, genus lactubacillus bacterium is Lactobacillus gasseri OLL2716 (preserving number: FERMBP-6999).。
[5] food compositions, it contains the gastrin formation inhibitor according to any one of aforementioned [1] ~ [4].
[6] food compositions according to aforementioned [5], it is the food with lactobacillus-fermented.
[7] food compositions according to aforementioned [6] wherein, is fermented-milk with the food of lactobacillus-fermented.
[8] food compositions according to aforementioned [7], wherein, fermented-milk is Yoghourt.
the effect of invention
Gastrin formation inhibitor of the present invention is owing to employing the dead thalline of genus lactubacillus bacterium, therefore not there is degermation (such as to Hp, patent documentation 1 and 2), the cell thus gastric acid secretion inhibiting that form stomach-tissue can be directly acted on.And then, also can expect its to such as remove the hyperchlorhydria after Hp, the reflux esophagitis caused thus and then and Hp have nothing to do gastritis, stomach ulcer, reflux esophagitis, such as increase with the age and the effect of reflux esophagitis caused etc.
Especially, according to the present invention, provide a kind of novel medicine or food, it does not utilize the anti-Hp effect of milk-acid bacteria, and prevention improves the series of symptoms, reflux esophagitis, the esophageal carcinoma etc. of the pyrosis, stomachache etc. that hyperchlorhydria causes.
And then such as, when the problem of gastritis that hydrochloric acid in gastric juice causes, reflux esophagitis appears in non-steroidal antiphlogiston (NSAID) user, present situation is, takes to utilize proton pump inhibitor (PPI) to carry out the methods for the treatment of of gastric acid secretion inhibiting.But, can time limit the longest of administration PPI be 8 weeks, and point of application be proton pump, therefore there is the side effect of the improper rising of expression of gastric acid secretion Cu Jin hormone and gastrin.On the other hand, measurable: gastrin formation inhibitor of the present invention is owing to directly acting on the cell of formation stomach-tissue to suppress the secretion of gastrin, therefore the side effect of the consumption of PPI or the gastrin rising after suppressing PPI administration to terminate can be reduced, can think, by combinationally using gastrin formation inhibitor of the present invention and PPI, gastritis, stomach ulcer, reflux esophagitis and hyperchlorhydria that hyperchlorhydria causes effectively can be improved.
In addition, owing to using dead thalline, temperature when not needing to regulate the traffic etc., therefore easy and simple to handle, without the anxiety of quality variation.In addition, do not need to consider the symbiotic relationship of itself and other microorganism (especially bacterium), unrestricted in relatively utilizing.
And then the dead thalline comprised in the food compositions of gastrin formation inhibitor energetically containing genus lactubacillus bacterium of the present invention, has diverse composition with food compositions in the past, by suppressing gastrin and can gastric acid secretion inhibiting etc.
Embodiment
Gastrin formation inhibitor of the present invention comprises the dead thalline that milk-acid bacteria (Lactobacillus) belongs to bacterium.Gastrin formation inhibitor of the present invention also can be used as medicine or foodstuff additive.In addition, gastrin formation inhibitor of the present invention can also comprise milk-acid bacteria (Lactobacillus) and belongs to the dead thalline of bacterium and do not comprise viable bacteria body.In addition, also dead thalline can only be comprised.
As the example of the bacterial classification that Lactobacillus belongs to, Lactobacillusdelbrueckiisubsp.burgalicus can be listed, Lactobacillusdelbrueckiisubsp.lactis, Lactobacillusparacaseisubsp.paracasei, Lactobacillushelveticus, Lactobacillushelveticussubsp.jugurti, Lactobacillusacidophilus, Lactobacilluscrispatus, Lactobacillusamylovorus, Lactobacillusgallinarum, Lactobacillusgasseri, Lactobacillusoris, Lactobacilluscaseisubsp.rhamnosus, Lactobacillusjohnsonii, Lactobacillusfermentum, Lactobacillusbrevis, as long as the generation that the genus lactubacillus bacterium in the present invention utilizes dead thalline to carry out preparation gastrin is just not particularly limited, also can be the one kind or two or more of previous example.Be preferably Lactobacillus gasseri, be particularly preferably Lactobacillus gasseri OLL2716 (preserving number: FERMBP-6999) (this bacterial strain is preserved in Independent Administrative Leged Industrial Technology Complex Inst's Patent Organism preservation center (life Advanced Industrial technical institute of old Govement Industrial Research Inst., Ministry of Commerce), east, Zhu Bo city, postcode 305-8566 Hitachinaka County, Japan 1 fourth order a kind of ground 1 central authorities the 6th (old address: east, Zhu Bo city, postcode 305-8566 Hitachinaka County, Japan 1 fourth order a kind No. 3) on May 24th, 1999).
When gastrin formation inhibitor of the present invention is used as medicine, as long as finally act on stomach as the dead thalline of the genus lactubacillus bacterium of effective constituent, then its route of administration is not particularly limited.The individuality of institute's administration suppresses the generation of gastrin, plays the effect improving hyperchlorhydria, gastritis, stomach ulcer, reflux esophagitis etc.
The administering mode of gastrin formation inhibitor of the present invention comprises oral administration and non-oral administration.Such as, also can be oral administration, through pipe administration.From the view point of easy and security, be preferably oral administration.In addition, formulation is not particularly limited, suitably can selects according to route of administration, such as, can list liquid preparation, capsule, granule, pill, clouding agent, emulsion, powder, tablet, syrup, injection, containing tablet etc.
As oral Preparation, can be made into known various formulation, such as, can be made into granule, powder, tablet, pill, capsule, liquid preparation, syrup, emulsion, clouding agent, containing tablets and other formulations.
As non-oral administration, can list and carry out administration with the form of injection etc.In addition, also can by gastrin formation inhibitor topical of the present invention to the region that will apply to process.Such as, can also be injected by the local in operation, use conduit to carry out administration.
As the carrier that can use in pharmaceutical composition of the present invention, the admissible carrier in medicine such as tensio-active agent, vehicle, colouring matter, spices, preservation material, stablizer, buffer reagent, clouding agent, isotonic agent, bonding agent, disintegrating agent, lubrication prescription, flow improver, correctives can be listed, suitably can use other conventional carrier.Specifically, light silicon anhydride, lactose, crystalline cellulose, N.F,USP MANNITOL, starch, calcium carboxymethylcellulose, Xylo-Mucine, hydroxypropylcellulose, HPMC, polyvinylacetal diethylamine, polyvinylpyrrolidone, gelatin, medium chain fatty acid Witepsol W-S 55, HCO60, white sugar, carboxymethyl cellulose, W-Gum, inorganic salts etc. can be listed.
When gastrin formation inhibitor of the present invention is used for medicine, the cell concentration of the dead thalline of contained genus lactubacillus bacterium is not particularly limited, preferably, when utilizing with the form of concentrated solution, is set to 2 × 10 10individual/more than g, is set to 1 × 10 when utilizing with the form of dry thing 11individual/more than g.
When gastrin inhibitor of the present invention is used as medicine, the use level of the dead thalline of genus lactubacillus bacterium is not particularly limited, and suitably can adjust according to formulation, symptom, body weight, purposes etc.
The odd-numbered day intake of medicine of the present invention is not particularly limited, and suitably can adjust according to age, symptom, body weight, purposes etc.Typical case, can absorb 0.1 ~ 10000mg/kg body weight, is preferably 1 ~ 2000mg/kg body weight, preferably absorbs 10 ~ 500mg/kg body weight further.In addition, acquisition method is not particularly limited, and can suitably change, and preparation ~ 1 daily ingestion 3 that such as 1 daily ingestion 1 is only approximately 1g is the preparation etc. of 10g.As the picked-up example of 1 day, such as, can exemplify people 1 bu 1 ~ 3 picked-up 1 ~ 5 × 10 of body weight 60kg 11the 6g club (stick) of the powder formulation of bacterium/g, every daily ingestion 100 ~ 300mg/kg.
Food compositions of the present invention contains gastrin formation inhibitor of the present invention.
Food compositions in the present invention is not particularly limited.Such as, can make the food originally just containing milk-acid bacteria, also can be the food originally not containing milk-acid bacteria.In addition, can also be liquid food (patient etc. can drink without the need to chewing into, the aqueous food with mobility).
Gastrin formation inhibitor of the present invention contains the dead thalline of genus lactubacillus bacterium,
Therefore food compositions of the present invention comprises the dead thalline of the one kind or two or more bacterium be selected from genus lactubacillus bacterium.The generation of the gastrin that described food compositions suppresses picked-up individual, plays the effect improving hyperchlorhydria, gastritis, stomach ulcer, reflux esophagitis etc.
Food compositions of the present invention can comprise saccharic, protein, lipid, vitamins, the required trace metal (manganous sulfate, zinc sulfate, magnesium chloride, salt of wormwood etc.) of organism, spices, other title complex.
As saccharic, carbohydrate, starch producing (except dextrin, Zulkovsky starch, Britain's starch, Sumstar 190, starch ester, starch ethers etc.), foodstuff fibre etc. can be listed.
As protein, include, for example out of animal or plant nature protein, their hydrolyzates such as whole milk powder, skim-milk, partially skimmed milk powder, casein, whey powder, whey protein, whey protein enriched material, whey protein sepd, alpha-casein, beta-casein, κ-casein, beta-lactoglobulin, alpha-lactalbumin, lactoferrin, soy-protein, egg protein, meat protein; The various milk-derived compositions etc. such as butter, milkiness mineral substance, missible oil, whey, amino acids, sialic acid, phosphatide, lactose.
As lipid, include, for example out lard, fish oil etc., their distillate oil, winterized stearin, transesterify wet goods animal raw fat; Plam oil, Thistle oil, Semen Maydis oil, rapeseed oil, Oleum Cocois, their distillate oil, winterized stearin, transesterify wet goods vegetative grease etc.
As vitamins, include, for example out vitamin A, carotenoid, vitamins B group, vitamins C, vitamins D group, vitamin-E, vitamin K group, vitamin P, CoenzymeQ10, nicotinic acid (niacin), nicotinic acid (nicotinicacid), pantothenic acid, vitamin H, inositol, choline, folic acid etc., as mineral substance class, include, for example out calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium etc.
Classification, the kind of food compositions of the present invention do not limit, can be functional foodstuff, specific food for health care, specific end use food, trophic function food, heath food, nursing food, in addition, also can be milk-product, the seasonings etc. such as dessert, lactobacillus drink, cheese, Yoghourt.The shape of beverage/food does not limit, solid fraction can be presented, the shape of all beverage/foods that aqueous, liquid food shape, g., jelly-like, sheet, particulate state, capsule shape etc. can circulate usually, can also add in various food (milk, refreshment drink, fermented-milk, Yoghourt, cheese, bread, biscuit, pretzel, Pizza, formula milk, liquid food, patient with food, nutritive food, frozen product, processed food and other delicatessen food etc.).The manufacture of above-mentioned beverage/food can be carried out according to the ordinary method of those skilled in the art.
The dead thalline of genus lactubacillus bacterium of the present invention can to add in the food with lactobacillus-fermented and to use.Such as fermented-milk, pickles, pickles etc. can be listed with the food of lactobacillus-fermented.Be particularly preferably fermented-milk, wherein preferred Yoghourt.With in the food of lactobacillus-fermented, the viable bacteria of genus lactubacillus bacterium is drawn growth curve and exponentially breeds functionality, viable count in aforesaid fermentation food naturally can be determined according to its applicable attenuation degree, in food compositions of the present invention, except viable bacteria body, also add dead thalline, thus the total amount contained (viable bacteria+dead bacterium) exceedes the thalline number comprised in the food of existing lactobacillus-fermented, effectively suppresses the generation of gastrin thus under one's belt.Such as, can infer, when Yoghourt, the biomass of the genus lactubacillus bacterium comprised in end article is average 100g is 1 × 10 10left and right, in order to effectively suppress the generation of gastrin, adds 1 × 10 8~ 5 × 10 13, preferably add 1 × 10 9~ 5 × 10 12, particularly preferably add 5 × 10 10~ 5 × 10 11the dead thalline of left and right, and make total amount (viable bacteria+dead bacterium) be 1 × 10 10~ 5 × 10 13left and right, preferably 1 × 10 10~ 5 × 10 12, be particularly preferably 1 × 10 11left and right.
The cell concentration of the dead thalline of the genus lactubacillus bacterium contained in food compositions of the present invention is not particularly limited, and preferably, is set to 2 × 10 when utilizing with the form of concentrated solution 10individual/more than g, is set to 3 × 10 when utilizing with the form of dry thing 11individual/more than g.
In food compositions of the present invention, the use level of the dead thalline of genus lactubacillus bacterium is not particularly limited, and suitably can adjust according to formulation, symptom, body weight, purposes etc.
The odd-numbered day intake of food compositions of the present invention is not particularly limited, and suitably can adjust according to age, symptom, body weight, purposes etc.Typical, 0.1 ~ 30000mg/kg body weight can be absorbed, preferably absorb 0.1 ~ 20000mg/kg body weight, preferably absorb 0.1 ~ 4000mg/kg body weight further.
Below, further illustrate the present invention based on embodiment, but described embodiment is illustration of the present invention, does not limit the present invention.
Embodiment
Test example: the gastrin employing the dead thalline of genus lactubacillus bacterium generates and suppresses
1. material and method
[bacterium]
Bacterium uses Lactobacillus gasseri OLL2716 (Mingzhi Dairy Co., Ltd).
In order to inoculate mouse, genus lactubacillus bacterium is bred 1 day in Difco (trade mark) MRSbroth (BectonDickinson).
The thermal treatment of genus lactubacillus bacterium by cultivating bacterial suspension 10 minutes and carrying out at 100 DEG C.
[mouse]
The BALB/c male mice of aseptic (germ-free, GF) is obtained from CLEAJapan, Inc..Use Trexler type pliability film plastic barrier device to give sterilized feed and water to female GF mouse, raise.
In order to make bacterium place occupy mouse, the viable bacteria of 109CFU being suspended in the phosphoric acid buffer (PBS) of 0.5ml, using the Mouse oral of stomach tube to 8 week age to inoculate 1 time.
As the process based on dead bacterium, by 10 10the heat treated bacterium (dead bacterium) of CFU is suspended in the phosphoric acid buffer (PBS) of 0.5ml, amounts to 10 every day and inoculates via stomach.
After 10 days that inoculate the last time, cut stomach, for immunohistochemical analysis.
[fabric analysis]
In order to carry out the histopathological analysis of stomach, the half of the stomach along the large curved excision of tail being fixed by 4% buffered formalin, is embedded in paraffin, cuts out the section of 2 μm.Then, according to standard method Hematorylin and eosin by section statining.The thickness of the muscle layer of stomach uses analysis software (OlympusBP2-BSW) to measure along Z-axis with microscope (OlympusAX80).Amount to the point that mensuration 10 place is different, obtain the mean thickness in each field of sample.Calculate the quantity of the core of the rectangle (150 × 200 μm) in the muscle layer in each field of tail.
In order to carry out immunohistochemical analysis, tissue slice being taken off paraffinized, seething with excitement 15 minutes in 10mM citrate buffer (pH6.0) with 98 DEG C with microwave oven, at room temperature close 15 minutes with the PBS containing 5% Normal Goat Serum.Section at 4 DEG C with antibody co-cultivation one Dinner, then, at room temperature with the secondary antibodies co-cultivation of carrying out marking with fluorescent mark 2 hours.Finally, sample DAPI (4,6-diamidino-2-phenylindone) and DABCO (Isosorbide-5-Nitrae-diazonium two ring [2,2,2] octane) processes.Section is carried out visual with the microscope (KeyenceBZ-9000) being adjusted to fluoroscopic image.Use AxioVissionRel.4.8 (Zeiss), morphological analysis is carried out to the section of dyeing to the gastrin positive (generation) cell of the random field at the interval of the 1mm of the axle along body of stomach-You Door hole length from mouse You Door hole.
Use and be diluted to the anti-gastrin antibody (rabbit polyclonal antibody, Dako) of 1:300, contrast and rabbit igg (Dako) and contrast and mouse IgG (Dako) as an antibody.Use goat anti-rabbit igg Alexa488 (molecular probe, MolecularProbe) and goat anti-mouse Alexa594 (molecular probe, MolecularProbe) as secondary antibodies.
[statistical analysis]
Result carries out statistical evaluation by suitably utilizing Student ' st inspection or one-way analysis of variance (ANOVA).The software program of SSPSversion16.0 (SSPS) or GraphPadPrism5 (GraphPadSoftware) is used for data analysis.
2. result
Measure without disposing GF mouse respectively, L.gasseriOLL2716 Su Ju limits bacterium (gnotobiote) mouse, PBS disposes GF mouse, administration imprisons gastrin founder cell quantity in Door hole through the stomach of the GF mouse of the dead thalline of heat treated L.gasseriOLL2716.The results are shown in following table.
[table 1]
Gastrin founder cell quantity in stomach pylorus hole
a)the founder cell quantity of 1mm in the horizontal direction of sample
b）Mean±SE
c)use stomach tube to the GF host mouse oral vaccination 1 time 10 in 8 week age 9the genus lactubacillus bacterium of CFU, and analyze after 10 days.
d)in the GF host mouse to 8 week age, 10 10before the dead thalline through heat treated genus lactubacillus bacterium of CFU is analyzed, in 10 days, every Nikkei stomach is inoculated.
*for GF or the GF/PBS group checked based on Turkey ' st, P<0.001
3. investigate
For oral administration through heat treated genus lactubacillus bacterium (dead thalline) GF mouse for, the minimizing quantity that can confirm gastrin founder cell is equal extent with the quantity derived by viable bacteria.Described remarkable minimizing also shows when Lactobacillus gasseri OLL2716.
utilizability in industry
According to gastrin inhibitor of the present invention, do not utilize the degermation to Hp, can directly act on the cell thus gastric acid secretion inhibiting that form stomach-tissue, gastritis, stomach ulcer and the reflux esophagitis that thus can cause as the non-Hp of improvement, improvement remove the pharmaceutical composition of the hyperchlorhydria after Hp etc.
And then, such as in food with lactobacillus-fermented etc., although in its food quality consider cannot make its contain a certain amount of more than viable bacteria, but according to gastrin inhibitor of the present invention, by using dead thalline, can use as the food compositions comprising the biomass (viable bacteria and dead bacterium) exceeded in existing food.

Claims (8)

1. a gastrin formation inhibitor, its comprise the dead thalline of genus lactubacillus bacterium as effective constituent, do not comprise viable bacteria body.
2. gastrin formation inhibitor according to claim 1, wherein, genus lactubacillus bacterium is Lactobacillus gasseri.
3. gastrin formation inhibitor according to claim 1, wherein, genus lactubacillus bacterium is Lactobacillus gasseri OLL2716 (preserving number: FERMBP-6999).
4. a food compositions, it contains the gastrin formation inhibitor according to any one of claims 1 to 3.
5. food compositions according to claim 4, it is the food with lactobacillus-fermented.
6. food compositions according to claim 5 wherein, is fermented-milk with the food of lactobacillus-fermented.
7. food compositions according to claim 6, wherein, fermented-milk is Yoghourt.
8. the dead thalline not comprising the genus lactubacillus bacterium of viable bacteria body is manufacturing the application in gastrin formation inhibitor.
CN201280003716.6A 2011-01-25 2012-01-24 Gastrin formation inhibitor and the food compositions containing this gastrin formation inhibitor Active CN103260632B (en)

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