CN105105145A - Lactobacillus plantarum and application thereof in preparing functional food for reducing blood glucose and blood fat - Google Patents
Lactobacillus plantarum and application thereof in preparing functional food for reducing blood glucose and blood fat Download PDFInfo
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- CN105105145A CN105105145A CN201510581420.8A CN201510581420A CN105105145A CN 105105145 A CN105105145 A CN 105105145A CN 201510581420 A CN201510581420 A CN 201510581420A CN 105105145 A CN105105145 A CN 105105145A
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- lactobacillus plantarum
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- diabetes
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- 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
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
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- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses lactobacillus plantarum and the application thereof in preparing functional food for reducing blood glucose and blood fat. The preservation and register number of the lactobacillus plantarum Sc52 is CGMCC NO.11027. The invention further discloses the application of the lactobacillus plantarum Sc52 in preparing a product. The invention further discloses a product. The active ingredient of the product is the lactobacillus plantarum Sc52 or lactobacillus plantarum Sc52 fungicide. The lactobacillus plantarum Sc52 is mainly used for reducing blood glucose and/or blood fat, promoting the growth of probiotics in the intestinal flora and/or restraining the growth of pathogenic bacteria in the intestinal flora. The lactobacillus plantarum Sc52 can serve as a functional probiotic to be applied to food, healthcare products and drugs and has broad industrial prospects.
Description
Technical field
The present invention relates to a lactobacillus plantarum and the application in the functional food of the hypoglycemic blood fat of preparation thereof.
Background technology
In recent years, the metabolic disorder that discovery feature food can improve blood pressure and blood lipoid is studied.Extensively be distributed in the phytosterol in vegetable oil, seed and nut, cereal and beans, its chemical constitution is similar to cholesterol structure, weaken cholesterol absorption by competitive inhibitory effect, and effectively can reduce T-CHOL and LDL-C content in serum.Soluble dietary fiber in the cereal such as oat, wheat, corn can reduce blood sugar level, improves Glucose tolerance test, can promote bile acid biosynthesis simultaneously, reduces LDL-C synthesis.Soybean protein in soymilk, bean curd can reduce human low density lipoprotein cholesterol and content of triglyceride.
Be different from normal food, functional food is for specific crowd designs the food with security and specific health care produced.Mainly through diet, Disease-causing gene is regulated, thus reach the symptom and the state of an illness of improving relevant disease, delay and prevent generation and the development of complication.Functional food has wide material sources, and toxic and side effect is little, and lower-price characteristic is easily accepted, and is daily health caring, regulates healthy best means.
Probio is an anthropoid microecologic regulator, intestinal microflora can be improved, promote the propagation of beneficial bacterium in enteron aisle, suppress the growth of harmful bacteria, eliminate carcinogen, improve immunity of organisms etc., for the control important in inhibiting of hypertension, high fat of blood, heart disease, diabetes and cancer.At present to probio as functional food improve hyperglycaemia, high fat of blood report less, thus the Application and Development of probio reducing blood sugar and blood fat will become a study hotspot of functional food, and will become the key areas of functional food development.
Diabetes (DiabetesMellitus, DM) be the internal system metabolic disturbance diseases of a class complexity, along with the increase of the incidence of disease, the ratio in diabetic shared by diabetes B, more than 90%, is the worldwide public health problem of serious threat human health.Diabetes B patient effectively can not utilize insulin or form insulin resistant, and then causes the metabolism system such as sugar, fat, protein in body disorderly, to cause organ dysfunction and exhaustion.Clinical treatment diabetes are based on oral insulin sensitizer, Drugs Promoting Insulin Secretion and alpha-glucosidase inhibitor, but these medicines of Long-Time Service can produce the serious side effect such as abdominal distension, inflatable, diarrhoea.
Summary of the invention
The object of this invention is to provide a lactobacillus plantarum and the application in the functional food of the hypoglycemic blood fat of preparation thereof.
Lactobacillus plantarum provided by the invention (Lactobacillusplantarum) Sc52, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 06 29th, 2015 and (be called for short CGMCC, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCCNO.11027.Lactobacillus plantarum (Lactobacillusplantarum) Sc52CGMCCNO.11027, is called for short Lactobacillus plantarum Sc52.
The present invention goes back the application of protective plant lactobacillus Sc52 in preparing product.
The function of described product be following (a1) at least one item in (a24):
(a1) hypoglycemic and/or reducing blood lipid;
(a2) promote the growth of probiotics in gut flora and/or suppress the pathogenic bacteria growth in gut flora;
(a3) blood-sugar content in serum is reduced;
(a4) lipids contents in serum is reduced;
(a5) total cholesterol level in serum is reduced;
(a6) content of triglyceride in serum is reduced;
(a7) fasting blood sugar is reduced;
(a8) GI is improved;
(a9) disorders of lipid metabolism is improved;
(a10) diabetes are treated and/or prevented;
(a11) blood-sugar content in Diabetes Mellitus is reduced;
(a12) lipids contents in Diabetes Mellitus is reduced;
(a13) total cholesterol level in Diabetes Mellitus is reduced;
(a14) content of triglyceride in Diabetes Mellitus is reduced;
(a15) fasting blood sugar of diabetic is reduced;
(a16) endotoxin content in Diabetes Mellitus is reduced;
(a17) insulin content in Diabetes Mellitus is raised;
(a18) levels of tnf-alpha in Diabetes Mellitus is reduced;
(a19) Content of IL-6 in Diabetes Mellitus is reduced;
(a20) promote the growth of probiotics in intestinal flora of patients with diabetes and/or suppress the pathogenic bacteria growth in intestinal flora of patients with diabetes;
(a21) GI of diabetic is improved;
(a22) disorders of lipid metabolism of diabetic is improved;
(a23) the low grade inflammation symptom of diabetic is improved;
(a24) hyperlipidemic conditions of diabetic is improved.
In described (a2), described probio specifically can be lactobacillus and/or Bifidobacterium.
In described (a2), described pathogenic bacteria specifically can be enterobacteria and/or enterococcus and/or C.perfringens.
In described (a20), described probio specifically can be lactobacillus and/or Bifidobacterium.
In described (a20), described pathogenic bacteria specifically can be enterobacteria and/or enterococcus and/or C.perfringens.
Described (a10) and/or described (a11) and/or described (a12) and/or described (a13) and/or described (a14) and/or described (a15) and/or described (a16) and/or described (a17) and/or described (a18) and/or described (a19) and/or described (a20) and/or described (a21) and/or described (a22) and/or described (a23) and/or described (in a24) described diabetes are diabetes B.
Described product can be food, health products or medicine.Described food specifically can be functional food, such as functional yoghourt, functional microbial inoculum etc.
The present invention also protects a kind of product, and its active component is the microbial inoculum of Lactobacillus plantarum Sc52 or Lactobacillus plantarum Sc52.In the microbial inoculum of described Lactobacillus plantarum Sc52, Lactobacillus plantarum Sc52 concentration is 10
8more than CFU/g, specifically can be 10
10-10
11cFU/g.The microbial inoculum of described Lactobacillus plantarum Sc52 can be the pulvis obtained with the bacterium solution preparation containing described Lactobacillus plantarum Sc52.The method preparing pulvis can be freeze drying or other technique.
The function of described product be following (a1) at least one item in (a24):
(a1) hypoglycemic and/or reducing blood lipid;
(a2) promote the growth of probiotics in gut flora and/or suppress the pathogenic bacteria growth in gut flora;
(a3) blood-sugar content in serum is reduced;
(a4) lipids contents in serum is reduced;
(a5) total cholesterol level in serum is reduced;
(a6) content of triglyceride in serum is reduced;
(a7) fasting blood sugar is reduced;
(a8) GI is improved;
(a9) disorders of lipid metabolism is improved;
(a10) diabetes are treated and/or prevented;
(a11) blood-sugar content in Diabetes Mellitus is reduced;
(a12) lipids contents in Diabetes Mellitus is reduced;
(a13) total cholesterol level in Diabetes Mellitus is reduced;
(a14) content of triglyceride in Diabetes Mellitus is reduced;
(a15) fasting blood sugar of diabetic is reduced;
(a16) endotoxin content in Diabetes Mellitus is reduced;
(a17) insulin content in Diabetes Mellitus is raised;
(a18) levels of tnf-alpha in Diabetes Mellitus is reduced;
(a19) Content of IL-6 in Diabetes Mellitus is reduced;
(a20) promote the growth of probiotics in intestinal flora of patients with diabetes and/or suppress the pathogenic bacteria growth in intestinal flora of patients with diabetes;
(a21) GI of diabetic is improved;
(a22) disorders of lipid metabolism of diabetic is improved;
(a23) the low grade inflammation symptom of diabetic is improved;
(a24) hyperlipidemic conditions of diabetic is improved.
In described (a2), described probio specifically can be lactobacillus and/or Bifidobacterium.
In described (a2), described pathogenic bacteria specifically can be enterobacteria and/or enterococcus and/or C.perfringens.
In described (a20), described probio specifically can be lactobacillus and/or Bifidobacterium.
In described (a20), described pathogenic bacteria specifically can be enterobacteria and/or enterococcus and/or C.perfringens.
Described (a10) and/or described (a11) and/or described (a12) and/or described (a13) and/or described (a14) and/or described (a15) and/or described (a16) and/or described (a17) and/or described (a18) and/or described (a19) and/or described (a20) and/or described (a21) and/or described (a22) and/or described (a23) and/or described (in a24) described diabetes are diabetes B.
Described product can be food, health products or medicine.Described food specifically can be functional food, such as functional yoghourt, functional microbial inoculum etc.
The present invention, to be separated the lactic acid bacteria of identifying for research object from traditional fermented food, obtains a strains of lactic acid bacteria new strains through a large amount of experiment screenings, called after Lactobacillus plantarum Sc52.Lactobacillus plantarum Sc52 has probiotic properties.By inducing the diabetes B model mice gavage obtained to give Lactobacillus plantarum Sc52 continuous 4 weeks to high lipid food in conjunction with Streptozotocin in the present invention, inquire into Lactobacillus plantarum Sc52 to the therapeutic action of diabetes B model mice and mechanism thereof.Result of study shows, Lactobacillus plantarum Sc52 can hypoglycemic, reducing blood lipid, reduction fasting blood sugar, improve glucose-tolerant, reduce serum cholesterol, regulate disorders of lipid metabolism, reduce serum endotoxin, reduce proinflammatory cytokine, thus improve low grade inflammation state and insulin resistance.
Lactobacillus plantarum Sc52 provided by the invention can be applied to food, health products and medicine field as functional probio, has extraordinary industrial prospect.
Accompanying drawing explanation
Fig. 1 is the testing result of carbohydrate tolerance test.
Fig. 2 is the testing result of gut flora.
Detailed description of the invention
Following embodiment is convenient to understand the present invention better, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is conventional method.Test material used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples, all arranges and repeats experiment for three times, results averaged.
C57BL/6J mouse (body weight 15-17g, 4-5 age in week): Jilin University's animal experimental center.
Solid BCP culture medium: solvent is water, containing yeast extract 2.5g/L, peptone 5g/L, glucose 5g/L, bromocresol purple 0.04g/L, agar 15g/L; PH7.0.The difference of liquid B CP culture medium and solid BCP culture medium is only not add agar.
Solid MRS culture medium: solvent is water, containing peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, KH
2pO
42g/L, sodium acetate 5g/L, natrium citricum 5g/L, MgSO
47H
2o0.2g/L, MnSO
44H
2o0.05g/L, Tween-80 1mL/L, agar 15g/L, glucose 20g/L; PH6.6.The difference of liquid MRS culture medium and solid MRS culture medium is only not add agar.
Lactobacillus culture medium (LBS): solvent is water, containing pancreas casein peptone 10.0g/L, yeast leaching powder 5.0g/L, potassium dihydrogen phosphate 6.0g/L, ferrous sulfate 0.034g/L, magnesium sulfate 0.575g/L, glucose 20.0g/L, sodium acetate 25.0g/L, ammonium citrate 2.0g/L, manganese sulfate 0.12g/L, Tween-80 1mL/L, agar 12g/L; PH5.5 ± 0.2.
Medium of Bifidobacterium (MRS+NNLP): solvent is water, containing yeast leaching powder 5.0g/L, peptone 10.0g/L, beef extract powder 10.0g/L, glucose 20.0g/L, dipotassium hydrogen phosphate 2.0g/L, natrium citricum 5.0g/L, sodium acetate 5.0g/L, magnesium sulfate 0.2g/L, manganese sulfate 0.05g/L, agar 12.0g/L, Tween-80 1mL/L, acidum nalidixicum 15mg/L, neomycinsulphate 100mg/L, paromomycin sulfate 200mg/L, lithium chloride 3g/L, L-cysteine hydrochloride 500mg/L; PH6.5 ± 0.2,
Enterobacteria culture medium (VRBDA): solvent is water, containing dusty yeast 3.0g/L, peptone 7.0g/L, sodium chloride 5.0g/L, glucose 10.0g/L, cholate 1.5g/L, crystal violet 0.002g/L, dimethyl diaminophenazine chloride 0.03g/L, agar 12g/L; PH7.3 ± 0.2.
Enterococcus culture medium (BEA): solvent is water, containing beef extract powder 3.0g/L, pancreas casein peptone 17.0g/L, dusty yeast 5.0g/L, bilein 10.0g/L, sodium chloride 5.0g/L, aesculin 1.0g/L, ferric citrate 0.5g/L, sodium azide 0.2g/L, natrium citricum 1.0g/L, agar 13.5g/L; PH7.1 ± 0.2.
C.perfringens culture medium (TSC): solvent is water, containing tryptone 15.0g/L, soya peptone 5.0g/L, yeast extract 5.0g/L, sodium metabisulfite 1.0g/L, ferric citrate 1.0g/L, agar 20.0g/L, pH7.6 ± 0.1, before toppling over plate, in every 250mL culture medium (50 DEG C), add the 20mL0.5%D-seromycin aqueous solution.
Basal feed: Jiangsu Province works in coordination with medical bioengineering Co., Ltd.
High lipid food (% represents g/100g): basal feed 75%, lard 10%, yolk powder 10%, cholesterol (food-grade, Zhengzhou Cang Yu chemical products Co., Ltd) 5%.
Carbohydrate tolerance test: after 12h, gavage gives D/W (dosage that gives of glucose is: 2g/kg body weight) on an empty stomach, respectively 0,30,60,90, measure blood glucose value after 120min, calculate Area under the curve of blood glucose (AreaUndertheCurve, AUC).
Employing bright of Shanghai mouse T-CHOL enzyme linked immunological kit (BP-E20095) detects T-CHOL (TotalCholesterol, the TC) content in serum.Employing bright of Shanghai mouse triglycerides enzyme linked immunological kit (BP-E20754) detects triglycerides (Triglyceride, the TG) content in serum.Employing bright of Shanghai mouse HDL-C enzyme linked immunological kit (BP-E20192) detects HDL-C (HighDensityLipoprotein, the HDL-C) content in serum.Employing bright of Shanghai mouse LDL-C enzyme linked immunological kit (BP-E20100) detects LDL-C (LowDensityLipoprotein, the LDL-C) content in serum.Bright of employing Shanghai mouse islets element enzyme linked immunological kit (BP-E20353) detects insulin (Insulin, the INS) content in serum.Employing bright of the Shanghai total enzyme linked immunological kit of Mouse endotoxin (BP-E200316) detects endotoxin (Endotoxin, the ET) content in serum.Employing bright of Shanghai mouse anti-tumor necrosis factor-α enzyme linked immunological kit (BP-E20220) detects anti-tumor necrosis factor-α (TumorNecrosisFactor α, the TNF-α) content in serum.Employing bright of Shanghai mouse interleukin-6 enzyme linked immunological kit (BP-E20012) detects interleukin-6 (Interleukin-6, the IL-6) content in serum.
The separation of embodiment 1, bacterial strain, preservation
One, the separation of bacterial strain
Sample is drawn materials: traditional zymotic sauerkraut.
Repeatedly to rule cultivation through BCP culture medium flat plate, single colony inoculation that picking produces yellow circle continues cultivation on MRS culture medium flat plate, repeatedly to rule culture purified, obtain many strains Pure cultured spawn through MRS culture medium flat plate.
The strain inoculation of pure culture, to liquid MRS medium culture, then adds 20% glycerine ,-80 DEG C of Refrigerator stores.
Through Gram-positive, catalase test is negative, indole reaction is negative, hydrogen sulfide production test is negative, gelatin liquefaction test is negative, hydrolyzed starch negative and nitrate reduction test feminine gender filter out many strains lactobacillus from the bacterial classification of pure culture.To wherein 1 lactobacillus plantarum strain called after Sc52.
Two, the qualification of bacterial strain
The Physiology and biochemistry qualification result of Sc52: Gram-positive, negative catalase; The bacillus do not moved; Can grow 15 DEG C and 45 DEG C; Tolerance 6.5%NaCl; Not hydrolyzed starch, not liquefy gelatin, do not produce hydrogen sulfide; Glucose fermentation produces acid not aerogenesis; Benzidine test is negative, and indole test is negative, and voges-Proskauer test is positive.
Sc52 is in even turbid growth in MRS fluid nutrient medium, and being long placed in thalline is white precipitate.
The optimum growth temperature of Sc52 37 ~ 42 DEG C, appropriate pH is 5.0 ~ 7.0.
API50CH (the French Mei Liai company) qualification result of Sc52: ribose, galactolipin, glucose, fructose, wood sugar, mannose, sweet mellow wine, sorbierite, amarogentin, ursin, N-acetyl-gucosamine, seven leaf-alcohols, salicin, cellobiose, synanthrin, lactose, melibiose, sucrose, trehalose, melezitose, raffinose, geraniol, D-R alcohol, gluconate can be utilized; Adonite, arabinose, sorbose, dulcitol, inositol, Alpha-Methyl-D-Glucose glycosides, Alpha-Methyl-D-MANNOSE glycosides, starch, glycogen, rhamnose, xylitol, D-turanose, D-Tag, D-rice threose, maltose, gluconate, rock sugar, Arabinose alcohol, 2-keto-D-gluconate salt, 5-keto-D-gluconate salt can not be utilized.
Qualification result shows, bacterial strain Sc52 is Lactobacillus plantarum (Lactobacillusplantarum).
Three, the preservation of bacterial strain
Lactobacillus plantarum (Lactobacillusplantarum) Sc52, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 06 29th, 2015 and (be called for short CGMCC, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preservation registration number is CGMCCNO.11027.Lactobacillus plantarum (Lactobacillusplantarum) Sc52CGMCCNO.11027, is called for short Lactobacillus plantarum Sc52.
Embodiment 2, Lactobacillus plantarum Sc52 are to the regulating action of blood sugar
One, packet transaction mode
80 male C 57 BL/6 J mouses, are divided into 4 groups at random, and often organize 20, packet transaction is as follows:
Blank group: all adopt normal diet to feed from on-test to end; 9th week, mouse is disposable celiac injecting normal saline 30mg/kg after 12 hours on an empty stomach; 10th carries out administration in thoughtful 13rd week, and dosage is " every kg body weight gives 12mL sterile PBS buffer at every turn ";
Model group: the 1st feeds with high lipid food for thoughtful 8 weeks, and the 9th thoughtful off-test normal diet is fed; 9th week, mouse is disposable celiac injection Streptozotocin 30mg/kg after 12 hours on an empty stomach; 10th carries out administration in thoughtful 13rd week; Dosage is " every kg body weight gives 12mL sterile PBS buffer at every turn ";
Positive controls: the 1st feeds with high lipid food for thoughtful 8 weeks, and the 9th thoughtful off-test normal diet is fed; 9th week, mouse is disposable celiac injection Streptozotocin 30mg/kg after 12 hours on an empty stomach; 10th carries out administration in thoughtful 13rd week; Dosage is for " every kg body weight gives 12mL melbine PBS solution, containing 120mg melbine in 12mL melbine solution at every turn; The solvent PBS buffer solution of melbine solution ";
Sc52 group: the 1st feeds with high lipid food for thoughtful 8 weeks, and the 9th thoughtful off-test normal diet is fed; 9th week, mouse is disposable celiac injection Streptozotocin 30mg/kg after 12 hours on an empty stomach; 10th carries out administration in thoughtful 13rd week; Dosage is for " every kg body weight gives 12mL bacterium liquid at every turn; The preparation method of bacterium liquid: with PBS buffer solution suspension plant lactobacillus Sc52, obtaining Lactobacillus plantarum Sc52 concentration is 1.0 × 10
9the bacterium liquid of CFU/mL ".
Namely 1st test 1-56 days in thoughtful 8 weeks, and namely the 10th test 64-91 days in thoughtful 13 weeks, by that analogy.Lumbar injection Streptozotocin or physiological saline carry out in test on the 57th day.
Two, the diabetic mice set up is judged whether
9th week (testing the 63rd day), tail vein blood measures mouse fasting blood sugar (FBG).The mouse meeting following standard is diabetes B model mice: double mensuration FBG >=11.1mmol/L.Result shows, test the 57th day, model group, positive controls and each mouse of Sc52 group are diabetes B model mice.
Three, blood sugar test
Respectively at the 10th week, the 11st week, the 12nd week, the 13rd week and the 14th week, tail vein blood measured mouse fasting blood sugar (every day is detected, and calculates the mean value in this week).Test and carry out carbohydrate tolerance test on the 99th day.
Fasting blood sugar the results are shown in Table 1.Compared with model group, Sc52 group mouse fasting blood sugar is on a declining curve.Sc52 group mouse fasting blood sugar significantly declined at the 12nd week, the 13rd week, the 14th week, and when the 14th week, blood glucose value drops to 8.45mmol/L, all has pole significant difference (P < 0.01) compared with model group mouse.Result shows, the hyperglycemia of Lactobacillus plantarum Sc52 to diabetes B has therapeutic action, significantly can reduce fasting blood sugar.
Table 1 Lactobacillus plantarum Sc52 is on the impact of diabetes B mouse blood sugar
Note: compare with model group: * has significant difference (P < 0.05), and * * has pole significant difference (P < 0.01).
Blood glucose value in carbohydrate tolerance test the results are shown in Figure 1A.Each group of mouse blood sugar value 30min after gavage glucose reaches peak value.Sc52 group mouse blood sugar value decrease speed is comparatively even, and during 120min, blood sugar amount drops to 10.72mmol/L, and there were significant differences compared with model group mouse (P < 0.05).Area under the curve of blood glucose in carbohydrate tolerance test is shown in Figure 1B.Sc52 group mouse blood sugar TG-AUC is starkly lower than model group mouse, and otherness extremely significantly (P < 0.01).Result shows, Lactobacillus plantarum Sc52 obviously can improve the situation of the GI of diabetes B, and effect is suitable with melbine.
Four, the detection of blood fat, insulin, endotoxin and inflammatory factor
(test the 100th day) during off-test, get each group of mouse, pluck eyeball and get blood, room temperature solidifies the centrifugal 15min of 3000r/min after 60min, collects supernatant (serum).Detect the content of T-CHOL, triglycerides, HDL-C, LDL-C, insulin, endotoxin, anti-tumor necrosis factor-α and the interleukin-6 in serum.
In serum, the testing result of blood fat correlation factor is in table 2.In model group mice serum, TC, TG, LDL-C content is all apparently higher than blank group, and HDL-C content is starkly lower than blank group, and this is because diabetes cause Mice Body intracellular metabolite disorderly.Compared with model group, in Sc52 group mice serum, TC, TG content declines.Result shows, Lactobacillus plantarum Sc52 obviously can improve the hyperlipidemic conditions of diabetes B.
Table 2 Lactobacillus plantarum Sc52 is on the impact of diabetes B lipid of mice
Note: compare with model group: * has significant difference (P < 0.05), and * * has pole significant difference (P < 0.01).
The testing result of serum insulin, endotoxin, inflammatory factor is in table 3.Diabetes B can cause chronic inflammation in Mice Body, improves TNF-alpha content and IL-6 content in serum.Compared with model group, in Sc52 group mice serum, TNF-alpha content has pole conspicuousness to decline (P < 0.01).Compared with model group, in Sc52 group mice serum, INS content rises, and ET level obviously declines, and otherness extremely significantly (P < 0.01).Result shows, Lactobacillus plantarum Sc52 can improve INS, ET level in diabetes B mouse in various degree, alleviates body chronic inflammation, reduces TNF-alpha content, reduces IL-6 content.
Table 3 Lactobacillus plantarum Sc52 is on the impact of insulin, endotoxin, inflammatory factor in diabetes B mice serum
| Group | INS(pmol/L) | ET(EU/L) | TNF-α(pg/mL) | IL-6(pg/mL) |
| Blank group | 57.48±8.55 | 60.23±5.35 | 304.45±23.07 | 264.16±27.85 |
| Model group | 28.64±3.29 | 83.05±7.41 | 383.66±35.85 | 327.49±29.68 |
| Positive controls | 45.66±6.21 ** | 64.54±5.28 ** | 288.17±21.06 ** | 255.23±31.42 ** |
| Sc52 group | 35.46±2.67 | 63.68±6.07 ** | 242.18±22.14 ** | 304.92±26.87 |
Note: compare with model group: * has significant difference (P < 0.05), and * * has pole significant difference (P < 0.01).
Embodiment 3, Lactobacillus plantarum Sc52 are to the regulating action of gut flora
One, packet transaction mode
48 male C 57 BL/6 J mouses, are divided into 4 groups at random, often organize 12.
Packet processing method is with the step one of embodiment 2.
Two, the detection of gut flora
Respectively at the 10th week (testing the 70th day), the 12nd week (testing the 84th day), the 14th week (testing the 98th day) and the 15th week (testing the 105th day), often group gets 3 mouse, de-neck is put to death, get mouse shaping intestinal contents under aseptic condition in vitro, the method for observing with adopting slat chain conveyor combining form after SPSS gradient dilution detects gut flora.
For detecting the identification mark of the culture medium of various gut flora, cultivation temperature, incubation time and various gut flora in table 4.
Table 4 culture medium, cultivation temperature, incubation time and identification mark
The results are shown in Figure 2.High lipid food causes mouse intestinal flora to lack of proper care after building mouse diabetes B model in conjunction with Streptozotocin simultaneously.In Sc52 group mouse intestinal, the quantity of lactobacillus and Bifidobacterium increases gradually along with the prolongation of gavage time, within the 14th week, substantially returns to blank group level, after stopping gastric infusion, in mouse intestinal lactobacillus and Bifidobacterium quantity slightly decline.The quantity of enterobacteria in Sc52 group mouse intestinal, enterococcus and C.perfringens declines gradually along with the prolongation for the treatment of time, and within the 14th week, substantially return to blank group level, after stopping gastric infusion, Microflora does not have obvious Long-term change trend.Above result shows, Lactobacillus plantarum Sc52 adds the main probio quantity of lactobacillus, Bifidobacterium two kinds, forms biological barrier together with other anaerobic bacteria flora, suppresses the field planting of the pathogenic bacteria such as enterobacteria, enterococcus, adjustment intestinal microbial balance, promotes that body is to healthy future development.
Claims (10)
1. the application of Lactobacillus plantarum (Lactobacillusplantarum) Sc52 in preparing product; The deposit number of described Lactobacillus plantarum (Lactobacillusplantarum) Sc52 is CGMCCNO.11027.
2. apply as claimed in claim 1, it is characterized in that: described product is food, health products or medicine.
3. a product, its active component is Lactobacillus plantarum (Lactobacillusplantarum) Sc52; The deposit number of described Lactobacillus plantarum (Lactobacillusplantarum) Sc52 is CGMCCNO.11027.
4. a product, its active component is the microbial inoculum of Lactobacillus plantarum (Lactobacillusplantarum) Sc52; The deposit number of described Lactobacillus plantarum (Lactobacillusplantarum) Sc52 is CGMCCNO.11027.
5. the product as described in claim 3 or 4, is characterized in that: described product is food, health products or medicine.
6. apply as claimed in claim 1 or 2, or the product as described in claim 3 or 4 or 5, is characterized in that: the function of described product be following (a1) at least one item in (a24):
(a1) hypoglycemic and/or reducing blood lipid;
(a2) promote the growth of probiotics in gut flora and/or suppress the pathogenic bacteria growth in gut flora;
(a3) blood-sugar content in serum is reduced;
(a4) lipids contents in serum is reduced;
(a5) total cholesterol level in serum is reduced;
(a6) content of triglyceride in serum is reduced;
(a7) fasting blood sugar is reduced;
(a8) GI is improved;
(a9) disorders of lipid metabolism is improved;
(a10) diabetes are treated and/or prevented;
(a11) blood-sugar content in Diabetes Mellitus is reduced;
(a12) lipids contents in Diabetes Mellitus is reduced;
(a13) total cholesterol level in Diabetes Mellitus is reduced;
(a14) content of triglyceride in Diabetes Mellitus is reduced;
(a15) fasting blood sugar of diabetic is reduced;
(a16) endotoxin content in Diabetes Mellitus is reduced;
(a17) insulin content in Diabetes Mellitus is raised;
(a18) levels of tnf-alpha in Diabetes Mellitus is reduced;
(a19) Content of IL-6 in Diabetes Mellitus is reduced;
(a20) promote the growth of probiotics in intestinal flora of patients with diabetes and/or suppress the pathogenic bacteria growth in intestinal flora of patients with diabetes;
(a21) GI of diabetic is improved;
(a22) disorders of lipid metabolism of diabetic is improved;
(a23) the low grade inflammation symptom of diabetic is improved;
(a24) hyperlipidemic conditions of diabetic is improved.
7. application as claimed in claim 6 or product, is characterized in that: in described (a2) and/or described (a20), described probio is lactobacillus and/or Bifidobacterium.
8. application as claimed in claims 6 or 7 or product, is characterized in that: in described (a2) and/or described (a20), and described pathogenic bacteria are enterobacteria and/or enterococcus and/or C.perfringens.
9. the application as described in claim 6 or 7 or 8 or product, is characterized in that: described diabetes are diabetes B.
10. Lactobacillus plantarum (Lactobacillusplantarum) Sc52, its deposit number is CGMCCNO.11027.
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| CN113308421A (en) * | 2021-07-30 | 2021-08-27 | 北京量化健康科技有限公司 | Lactobacillus plantarum BUFX and application thereof in metabolic syndrome |
| CN113308421B (en) * | 2021-07-30 | 2022-01-07 | 北京量化健康科技有限公司 | Lactobacillus plantarum BUFX and application thereof in metabolic syndrome |
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| CN114381395B (en) * | 2021-12-30 | 2023-06-06 | 杭州康源食品科技有限公司 | Lactobacillus plantarum ZJFFN 1 and application thereof |
| CN114317353B (en) * | 2021-12-30 | 2023-09-15 | 浙江大学 | Lactobacillus plantarum ZJFFYJ 7 and application thereof |
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