CN105105145B - Lactobacillus plantarum and application thereof in preparation of functional food for reducing blood sugar and blood fat - Google Patents

Lactobacillus plantarum and application thereof in preparation of functional food for reducing blood sugar and blood fat Download PDF

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CN105105145B
CN105105145B CN201510581420.8A CN201510581420A CN105105145B CN 105105145 B CN105105145 B CN 105105145B CN 201510581420 A CN201510581420 A CN 201510581420A CN 105105145 B CN105105145 B CN 105105145B
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lactobacillus plantarum
serum
reducing
blood
mice
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CN105105145A (en
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李盛钰
段翠翠
高磊
赵玉娟
牛春华
栾畅
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Ming Zhiyuan Bio Tech Ltd Of Jilin Province
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吉林省命之元生物科技有限公司
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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
    • 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
    • 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
    • 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

Abstract

The invention discloses lactobacillus plantarum and application thereof in preparing functional food for reducing blood sugar and blood fat. The lactobacillus plantarum Sc52 provided by the invention has a preservation registration number of CGMCC NO. 11027. The invention also protects the application of the lactobacillus plantarum Sc52 in preparing products. The invention also protects a product, and the active ingredient of the product is a microbial inoculum of lactobacillus plantarum Sc52 or lactobacillus plantarum Sc 52. The lactobacillus plantarum Sc52 provided by the invention has the main functions as follows: reducing blood glucose and/or blood lipid; promote the growth of probiotics in the intestinal flora and/or inhibit the growth of pathogenic bacteria in the intestinal flora. The lactobacillus plantarum Sc52 provided by the invention can be used as functional probiotics to be applied to the fields of food, health care products and medicines, and has good industrial prospects.

Description

Lactobacillus plantarum and application thereof in preparation of functional food for reducing blood sugar and blood fat
Technical Field
The invention relates to lactobacillus plantarum and application thereof in preparing functional food for reducing blood sugar and blood fat.
Background
In recent years, research finds that functional foods can improve abnormal metabolism of blood sugar and blood fat. Phytosterols, which are widely distributed in vegetable oils, seeds and nuts, grains and beans, have a chemical structure similar to that of cholesterol, can weaken cholesterol absorption by competitive inhibition, and can effectively reduce the total cholesterol and low density lipoprotein cholesterol content in serum. The soluble dietary fiber in cereals such as oat, wheat and corn can reduce blood sugar level, improve glucose tolerance, promote bile acid metabolism and reduce low density lipoprotein cholesterol synthesis. The soybean protein in soybean milk and bean curd can reduce the content of low density lipoprotein cholesterol and triglyceride in human body.
Unlike general foods, functional foods are foods designed and produced for specific people to have safety and specific health-care functions. The pathogenic gene is regulated mainly by diet, thereby achieving the purposes of improving symptoms and illness states of related diseases and delaying and preventing the occurrence and development of complications. The functional food has the characteristics of wide sources, small toxic and side effects, low price and the like, is easily accepted by people, and is the best means for daily health care and body health regulation.
The probiotics is a micro-ecological regulator for human bodies, can improve the structure of intestinal flora, promote the proliferation of beneficial bacteria in intestinal tracts, inhibit the growth of harmful bacteria, eliminate carcinogenic factors, improve the immunity of organisms and the like, and has important significance for preventing and treating hypertension, hyperlipidemia, heart disease, diabetes and cancers. At present, few reports are reported on probiotics as functional food for improving hyperglycemia and hyperlipidemia, so that the development and application of the probiotics for reducing blood sugar and blood fat become a research hotspot of the functional food and an important field of the development of the functional food.
Diabetes Mellitus (DM) is a complex disease of metabolic disorders of the endocrine system, and with increasing incidence, type 2 diabetes mellitus accounts for over 90% of patients with diabetes mellitus, which is a worldwide public health problem seriously threatening human health. Type 2 diabetic patients cannot effectively utilize insulin or develop insulin resistance, and further cause disorder of metabolic systems such as sugar, fat, protein, etc. in vivo, resulting in organ dysfunction and failure. Clinically, the diabetes is mainly treated by oral insulin sensitizer, insulin secretagogue and alpha-glycosidase inhibitor, but the drugs can cause severe side effects such as abdominal distension, flatulence, diarrhea and the like after being used for a long time.
Disclosure of Invention
The invention aims to provide lactobacillus plantarum and application thereof in preparing functional food for reducing blood sugar and blood fat.
The lactobacillus plantarum Sc52 provided by the invention has been preserved in China general microbiological culture Collection center (CGMCC for short, with the address of CGMCC No. 3 of the national institute of sciences, China) in 29.06.2015 in the Korean area, and the preservation registration number is CGMCC NO. 11027. Lactobacillus plantarum (Lactobacillus plantarum) Sc52CGMCC NO.11027, abbreviated as Lactobacillus plantarum Sc 52.
The invention also protects the application of the lactobacillus plantarum Sc52 in preparing products.
The product functions as at least one of the following (a1) to (a 24):
(a1) reducing blood glucose and/or blood lipid;
(a2) promoting the growth of probiotics in the intestinal flora and/or inhibiting the growth of pathogenic bacteria in the intestinal flora;
(a3) reducing blood glucose levels in serum;
(a4) reducing the blood lipid content in serum;
(a5) reducing total cholesterol content in serum;
(a6) reducing the triglyceride content in serum;
(a7) reducing fasting blood glucose;
(a8) improving glucose intolerance;
(a9) ameliorating a disorder of lipid metabolism;
(a10) treatment and/or prevention of diabetes;
(a11) reducing blood sugar content in blood serum of diabetic;
(a12) reducing blood lipid content in blood serum of diabetic;
(a13) reducing the total cholesterol content in the serum of a diabetic patient;
(a14) reducing the triglyceride content in the serum of a diabetic patient;
(a15) reducing fasting blood glucose value of diabetic;
(a16) reducing the endotoxin content in the serum of a diabetic patient;
(a17) increasing the insulin content in the serum of a diabetic patient;
(a18) reducing the content of tumor necrosis factor-alpha in blood serum of a diabetic patient;
(a19) reducing the content of interleukin-6 in the serum of a diabetic patient;
(a20) promoting the growth of probiotics in the intestinal flora of the diabetic and/or inhibiting the growth of pathogenic bacteria in the intestinal flora of the diabetic;
(a21) improving glucose intolerance in diabetic patients;
(a22) improving lipid metabolism disorder of diabetic patients;
(a23) improving low grade inflammation symptoms of diabetic patients;
(a24) improving the hyperlipemia symptoms of the diabetics.
In the (a2), the probiotic bacteria may be lactobacillus and/or bifidobacterium.
In the (a2), the pathogenic bacteria can be enterobacteria and/or enterococcus and/or clostridium perfringens.
In the (a20), the probiotic bacteria may be lactobacillus and/or bifidobacterium.
In the (a20), the pathogenic bacteria can be enterobacteria and/or enterococcus and/or clostridium perfringens.
Said (a10) and/or said (a11) and/or said (a12) and/or said (a13) and/or said (a14) and/or said (a15) and/or said (a16) and/or said (a17) and/or said (a18) and/or said (a19) and/or said (a20) and/or said (a21) and/or said (a22) and/or said (a23) and/or said (a24), said diabetes being type 2 diabetes.
The product may be a pharmaceutical product. The food can be functional food, such as functional yogurt, functional bacteria, etc.
The invention also protects a product, and the active ingredient of the product is a microbial inoculum of lactobacillus plantarum Sc52 or lactobacillus plantarum Sc 52. In the bacterial agent of the lactobacillus plantarum Sc52, the concentration of the lactobacillus plantarum Sc52 is more than 108CFU/g, and specifically can be 1010 and 1011 CFU/g. The bacterial agent of the lactobacillus plantarum Sc52 can be powder prepared by using bacterial liquid containing the lactobacillus plantarum Sc 52. The method of preparing the powder may be freeze-drying or other processes.
The product functions as at least one of the following (a1) to (a 24):
(a1) reducing blood glucose and/or blood lipid;
(a2) promoting the growth of probiotics in the intestinal flora and/or inhibiting the growth of pathogenic bacteria in the intestinal flora;
(a3) reducing blood glucose levels in serum;
(a4) reducing the blood lipid content in serum;
(a5) reducing total cholesterol content in serum;
(a6) reducing the triglyceride content in serum;
(a7) reducing fasting blood glucose;
(a8) improving glucose intolerance;
(a9) ameliorating a disorder of lipid metabolism;
(a10) treatment and/or prevention of diabetes;
(a11) reducing blood sugar content in blood serum of diabetic;
(a12) reducing blood lipid content in blood serum of diabetic;
(a13) reducing the total cholesterol content in the serum of a diabetic patient;
(a14) reducing the triglyceride content in the serum of a diabetic patient;
(a15) reducing fasting blood glucose value of diabetic;
(a16) reducing the endotoxin content in the serum of a diabetic patient;
(a17) increasing the insulin content in the serum of a diabetic patient;
(a18) reducing the content of tumor necrosis factor-alpha in blood serum of a diabetic patient;
(a19) reducing the content of interleukin-6 in the serum of a diabetic patient;
(a20) promoting the growth of probiotics in the intestinal flora of the diabetic and/or inhibiting the growth of pathogenic bacteria in the intestinal flora of the diabetic;
(a21) improving glucose intolerance in diabetic patients;
(a22) improving lipid metabolism disorder of diabetic patients;
(a23) improving low grade inflammation symptoms of diabetic patients;
(a24) improving the hyperlipemia symptoms of the diabetics.
In the (a2), the probiotic bacteria may be lactobacillus and/or bifidobacterium.
In the (a2), the pathogenic bacteria can be enterobacteria and/or enterococcus and/or clostridium perfringens.
In the (a20), the probiotic bacteria may be lactobacillus and/or bifidobacterium.
In the (a20), the pathogenic bacteria can be enterobacteria and/or enterococcus and/or clostridium perfringens.
Said (a10) and/or said (a11) and/or said (a12) and/or said (a13) and/or said (a14) and/or said (a15) and/or said (a16) and/or said (a17) and/or said (a18) and/or said (a19) and/or said (a20) and/or said (a21) and/or said (a22) and/or said (a23) and/or said (a24), said diabetes being type 2 diabetes.
The product may be a pharmaceutical product. The food can be functional food, such as functional yogurt, functional bacteria, etc.
The invention takes lactic acid bacteria separated and identified from traditional fermented food as research objects, and obtains a new lactic acid bacteria strain named as lactobacillus plantarum Sc52 through a large amount of experimental screening. The lactobacillus plantarum Sc52 has probiotic properties. In the invention, lactobacillus plantarum Sc52 is continuously fed to the type 2 diabetes model mouse obtained by inducing high-fat feed and streptozotocin through intragastric administration for 4 weeks, and the treatment effect and mechanism of the lactobacillus plantarum Sc52 on the type 2 diabetes model mouse are discussed. Research results show that the lactobacillus plantarum Sc52 can reduce blood sugar, blood fat and fasting blood glucose, improve glucose tolerance, reduce serum cholesterol, regulate lipid metabolism disorder, reduce serum endotoxin and proinflammatory cytokines, thereby improving low-grade inflammation state and insulin resistance.
The lactobacillus plantarum Sc52 provided by the invention can be used as functional probiotics to be applied to the field of medicines, and has a very good industrial prospect.
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FIG. 1 shows the results of the glucose tolerance test.
FIG. 2 shows the results of detection of intestinal flora.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
C57BL/6J mice (body weight 15-17g, 4-5 weeks old): jilin university animal experiment center.
Solid BCP medium: the solvent is water, and contains 2.5g/L yeast extract, 5g/L peptone, 5g/L glucose, 0.04g/L bromocresol purple and 15g/L agar; pH 7.0. Liquid BCP media differs from solid BCP media only in that no agar is added.
Solid MRS medium: the solvent is water, and contains 10g/L of peptone, 10g/L of beef extract, 5g/L, KH2PO42g/L of yeast extract, 5g/L of sodium acetate, 5g/L, MgSO 4.7 H2O0.2g/L, MnSO 4.4 H2O0.05g/L of sodium citrate, 801mL/L of tween-801, 15g/L of agar and 20g/L of glucose; pH 6.6. Liquid MRS medium differs from solid MRS medium only in that no agar is added.
Lactobacillus medium (LBS): the solvent is water, and contains 10.0g/L of tryptone, 5.0g/L of yeast extract powder, 6.0g/L of monopotassium phosphate, 0.034g/L of ferrous sulfate, 0.575g/L of magnesium sulfate, 20.0g/L of glucose, 25.0g/L of sodium acetate, 2.0g/L of ammonium citrate, 0.12g/L of manganese sulfate, Tween-801 mL/L and 12g/L of agar; pH5.5. + -. 0.2.
Bifidobacterium culture medium (MRS + NNLP): the solvent is water, and the yeast extract powder comprises 5.0g/L, peptone 10.0g/L, beef extract powder comprises 10.0g/L, glucose 20.0g/L, dipotassium hydrogen phosphate 2.0g/L, sodium citrate 5.0g/L, sodium acetate 5.0g/L, magnesium sulfate 0.2g/L, manganese sulfate 0.05g/L, agar 12.0g/L, Tween-801 mL/L, nalidixic acid 15mg/L, neomycin sulfate 100mg/L, paromomycin sulfate 200mg/L, and lithium chloride 3g/L, L-cysteine hydrochloride 500 mg/L; the pH value is 6.5 +/-0.2,
enterobacter medium (VRBDA): the solvent is water, and contains 3.0g/L of yeast powder, 7.0g/L of peptone, 5.0g/L of sodium chloride, 10.0g/L of glucose, 1.5g/L of bile salt, 0.002g/L of crystal violet, 0.03g/L of neutral red and 12g/L of agar; pH 7.3. + -. 0.2.
Enterococcus culture medium (BEA): the solvent is water, and contains 3.0g/L of beef extract powder, 17.0g/L of tryptone, 5.0g/L of yeast powder, 10.0g/L of ox gall powder, 5.0g/L of sodium chloride, 1.0g/L of esculin, 0.5g/L of ferric ammonium citrate, 0.2g/L of sodium azide, 1.0g/L of sodium citrate and 13.5g/L of agar; pH 7.1. + -. 0.2.
Clostridium perfringens medium (TSC): the solvent was water, 15.0g/L tryptone, 5.0g/L soytone, 5.0g/L yeast extract, 1.0g/L sodium metabisulfite, 1.0g/L ferric ammonium citrate, 20.0g/L agar, pH 7.6. + -. 0.1, 20mL of 0.5% aqueous solution of D-cycloserine was added per 250mL of the medium (50 ℃ C.) before pouring the plate.
Basic feed: jiangsu province, cooperative medical bioengineering, Limited liability company.
High-fat diet (% for g/100 g): 75% of basic feed, 10% of lard, 10% of yolk powder and 5% of cholesterol (food grade, Zhengzhou xanthium chemical product Co., Ltd.).
Sugar tolerance test: after an fasting period of 12 hours, an aqueous glucose solution (glucose dose: 2g/kg body weight) was administered by gavage, and the blood glucose values were measured after 0, 30, 60, 90, and 120min, respectively, to calculate the area under the blood glucose curve (AUC).
The Total Cholesterol (TC) content in serum is detected by adopting a Shanghai Langton mouse total cholesterol enzyme linked immunosorbent assay kit (BP-E20095). Triglyceride (TG) content in serum is detected by adopting Shanghai Langton mouse Triglyceride enzyme-linked immunoassay kit (BP-E20754). The content of high density Lipoprotein cholesterol (HDL-C) in serum is detected by adopting a Shanghai Langton mouse high density Lipoprotein cholesterol enzyme linked immunosorbent assay kit (BP-E20192). The content of low-density lipoprotein cholesterol (LDL-C) in serum is detected by adopting a Shanghai Langton mouse low-density lipoprotein cholesterol enzyme linked immunosorbent assay kit (BP-E20100). The content of Insulin (INSulin, INS) in serum is detected by adopting Shanghai Langton mouse Insulin enzyme-linked immunosorbent assay kit (BP-E20353). The Endotoxin (ET) content in serum is detected by adopting a Shanghai Langton mouse Endotoxin total enzyme linked immunosorbent assay kit (BP-E200316). The anti-tumor necrosis factor-alpha (TNF-alpha) content in serum is detected by adopting a Shanghai Langton mouse anti-tumor necrosis factor-alpha enzyme linked immunosorbent assay kit (BP-E20220). Detecting the content of Interleukin-6 (Interleukin-6, IL-6) in serum by adopting a Shanghai Langton mouse Interleukin-6 enzyme linked immunosorbent assay kit (BP-E20012).
Example 1 isolation, identification and preservation of strains
First, isolation of the Strain
Taking a sample: fermenting pickled Chinese cabbage traditionally.
And repeatedly carrying out streak culture by using a BCP (brain-derived plasma) culture medium plate, selecting a single colony producing a yellow circle, inoculating the single colony on an MRS (methicillin resistant Staphylococcus) culture medium plate for continuous culture, and repeatedly carrying out streak culture and purification by using the MRS culture medium plate to obtain a plurality of pure culture strains.
Inoculating the pure cultured strain to liquid MRS culture medium, culturing, adding 20% glycerol, and storing at-80 deg.C in refrigerator.
Multiple strains of lactobacillus were screened from the pure cultured strains by gram-positive staining, catalase test negative, indole reaction negative, hydrogen sulfide production test negative, gelatin liquefaction test negative, starch hydrolysis test negative, and nitrate reduction test negative. Among them, 1 strain of Lactobacillus plantarum was designated as Sc 52.
II, identification of the strains
Physiological and biochemical identification results of Sc 52: gram positive, catalase negative; immotile bacilli; capable of growing at 15 ℃ and 45 ℃; tolerance of 6.5% NaCl: starch is not hydrolyzed, gelatin is not liquefied, and hydrogen sulfide is not generated; the acid produced by fermenting glucose does not produce gas; benzidine test negative, indole test negative, and acetyl methyl methanol test positive.
Sc52 shows uniform turbid growth in MRS liquid culture medium, and the thalli is white precipitate after long-time placement.
The optimal growth temperature of the Sc52 is 37-42 ℃, and the pH is 5.0-7.0.
API50CH (Merrier, France) identification of Sc 52: ribose, galactose, glucose, fructose, xylose, mannose, mannitol, sorbitol, amygdalin, arbutin, N-acetyl-glucosamine, heptaphyllol, saligenin, cellobiose, inulin, lactose, melibiose, sucrose, trehalose, melezitose, raffinose, geraniol, D-arabitol, gluconate can be used; adonitol, arabinose, sorbose, dulcitol, inositol, alpha-methyl-D-glucoside, alpha-methyl-D-mannoside, starch, glycogen, rhamnose, xylitol, D-turanose, D-tagatose, D-stachyose, maltose, gluconate, fucose, L-arabitol, 2-keto-gluconate, 5-keto-gluconate cannot be utilized.
The identification result shows that the strain Sc52 is lactobacillus plantarum (Lactobacillus plantarum).
Third, preservation of the Strain
Lactobacillus plantarum Sc52 has been deposited in China general microbiological culture Collection center (CGMCC, address: institute of microbiology, China academy of sciences, No. 3, Xilu 1 on North Chen of the Korean district, Beijing) on 29.06.2015, and the collection registration number is CGMCC NO. 11027. Lactobacillus plantarum (Lactobacillus plantarum) Sc52CGMCC NO.11027, abbreviated as Lactobacillus plantarum Sc 52.
Example 2 blood sugar regulating action of Lactobacillus plantarum Sc52
One, grouping processing mode
80 male C57BL/6J mice, randomly divided into 4 groups of 20 mice each, were treated as follows:
blank control group: feeding the test by adopting common feed from the beginning to the end of the test; at the 9 th week, the mice are injected with 30mg/kg of physiological saline at one time in the abdominal cavity after fasting for 12 hours; administration was performed from week 10 to week 13 at a dose of "12 mL of sterile PBS buffer per kg body weight;
model group: feeding with high fat feed in 1-8 weeks, and feeding with common feed in 9-9 weeks; at 9 weeks, 30mg/kg of streptozotocin is injected into the abdominal cavity once after the mice fasting for 12 hours; administration is performed from week 10 to week 13; the dose was "12 mL of sterile PBS buffer per kg body weight;
positive control group: feeding with high fat feed in 1-8 weeks, and feeding with common feed in 9-9 weeks; at 9 weeks, 30mg/kg of streptozotocin is injected into the abdominal cavity once after the mice fasting for 12 hours; administration is performed from week 10 to week 13; the dose was "12 mL of metformin PBS solution per kg body weight, 12mL of metformin solution containing 120mg of metformin; solvent PBS buffer of metformin solution ";
sc52 group: feeding with high fat feed in 1-8 weeks, and feeding with common feed in 9-9 weeks; at 9 weeks, 30mg/kg of streptozotocin is injected into the abdominal cavity once after the mice fasting for 12 hours; administration is performed from week 10 to week 13; the administration dose is that the bacterial liquid is administered by 12mL per kilogram of body weight; the preparation method of the bacterial liquid comprises the following steps: the Lactobacillus plantarum Sc52 was suspended in PBS buffer to obtain a suspension containing Lactobacillus plantarum Sc52 at a concentration of 1.0X 109CFU/mL ".
Weeks 1 to 8, i.e., days 1-56 of the trial, weeks 10 to 13, i.e., days 64-91 of the trial, and so on. Intraperitoneal injection of streptozotocin or saline was performed on day 57 of the experiment.
II, judging whether the established diabetes model mouse is established
At week 9 (day 63 of the experiment), the mice were bled for Fasting Blood Glucose (FBG). The mice meeting the following criteria are type 2 diabetes model mice: FBG is measured for two times continuously and is more than or equal to 11.1 mmol/L. The result shows that on the 57 th day of the test, the model group mice, the positive control group mice and the Sc52 group mice are type 2 diabetes model mice.
Third, blood sugar detection
The fasting blood glucose level of the mice was measured by tail vein blood sampling at week 10, week 11, week 12, week 13 and week 14, respectively (the average value in the week was calculated by daily measurement). Test day 99 a glucose tolerance test was performed.
The fasting blood glucose results are shown in table 1. Compared with the model group, the fasting blood glucose value of the mice in the Sc52 group is in a descending trend. The fasting blood glucose values of the mice in the Sc52 group are remarkably reduced at 12 weeks, 13 weeks and 14 weeks, and the blood glucose value at the 14 week is reduced to 8.45mmol/L, which are very different from those of the mice in the model group (P is less than 0.01). The result shows that the lactobacillus plantarum Sc52 has a treatment effect on hyperglycemia of type 2 diabetes and can obviously reduce the fasting blood glucose value.
TABLE 1 Effect of Lactobacillus plantarum Sc52 on blood glucose in type 2 diabetic mice
Figure BFW0000023323610000081
Note: comparison with model groups: significant differences (P < 0.05) and very significant differences (P < 0.01).
The results of blood glucose values in the glucose tolerance test are shown in fig. 1A. The blood glucose values of the mice of each group reach the peak value 30min after the intragastric glucose administration. The blood sugar value of the Sc52 group mice is reduced at a uniform speed, the blood sugar value is reduced to 10.72mmol/L at 120min, and the blood sugar value is obviously different from that of the model group mice (P is less than 0.05). The area under the blood glucose curve in the glucose tolerance test is shown in fig. 1B. The area under the blood glucose curve of the Sc52 group mice is obviously lower than that of the model group mice, and the difference is very obvious (P < 0.01). The result shows that the lactobacillus plantarum Sc52 can obviously improve the condition of glucose intolerance of type 2 diabetes mellitus, and the effect is equivalent to that of metformin.
Detection of blood fat, insulin, endotoxin and inflammatory factor
At the end of the test (day 100), each group of mice was collected, the eyeballs were removed, blood was collected, the mixture was coagulated at room temperature for 60min and centrifuged at 3000r/min for 15min, and the supernatant (serum) was collected. Detecting the content of total cholesterol, triglyceride, high density lipoprotein cholesterol, low density lipoprotein cholesterol, insulin, endotoxin, anti-tumor necrosis factor-alpha and interleukin-6 in serum.
The results of the serum lipid-related factor measurements are shown in Table 2. The contents of TC, TG and LDL-C in the serum of the model group mouse are all obviously higher than those of a blank control group, and the content of HDL-C is obviously lower than that of the blank control group, which is caused by diabetes to cause metabolic disorder in the mouse body. Compared with the model group, the serum TC and TG content of the Sc52 group mice is reduced. The result shows that the lactobacillus plantarum Sc52 can obviously improve the hyperlipemia symptom of type 2 diabetes.
TABLE 2 influence of Lactobacillus plantarum Sc52 on blood lipids in type 2 diabetic mice
Figure BFW0000023323610000091
Note: comparison with model groups: significant differences (P < 0.05) and very significant differences (P < 0.01).
The results of the detection of insulin, endotoxin and inflammatory factor in serum are shown in Table 3. Type 2 diabetes causes chronic inflammation in mice, increasing the levels of TNF-alpha and IL-6 in serum. Compared with the model group, the serum content of TNF-alpha in the mice of the Sc52 group is reduced with great significance (P is less than 0.01). Compared with the model group, the serum INS content of the Sc52 mice is increased, the ET level is obviously reduced, and the difference is extremely obvious (P is less than 0.01). The results show that the lactobacillus plantarum Sc52 can improve INS and ET levels in type 2 diabetic mice to different degrees, relieve chronic inflammation of organisms, reduce TNF-alpha content and reduce IL-6 content.
TABLE 3 Effect of Lactobacillus plantarum Sc52 on insulin, endotoxin, inflammatory factors in serum of type 2 diabetic mice
Group of INS(pmol/L) ET(EU/L) TNF-α(pg/mL) IL-6(pg/mL)
Blank control group 57.48±8.55 60.23±5.35 304.45±23.07 264.16±27.85
Model set 28.64±3.29 83.05±7.41 383.66±35.85 327.49±29.68
Positive control group 45.66±6.21** 64.54±5.28** 288.17±21.06** 255.23±31.42**
Group Sc52 35.46±2.67 63.68±6.07** 242.18±22.14** 304.92±26.87
Note: comparison with model groups: significant differences (P < 0.05) and very significant differences (P < 0.01).
Example 3 Regulation of intestinal flora by Lactobacillus plantarum Sc52
One, grouping processing mode
48 male C57BL/6J mice, randomly divided into 4 groups of 12 mice each.
The packet processing method is the same as the first step of embodiment 2.
Second, detection of intestinal flora
At each of the 10 th week (test day 70), 12 th week (test day 84), 14 th week (test day 98) and 15 th week (test day 105), 3 mice were taken out of the test tube, sacrificed, and the contents of the shaped intestine of each mouse were aseptically taken out into the test tube, and the intestinal flora was examined by plate culture combined with morphological observation after dilution with a sterile physiological saline gradient.
The culture media, culture temperatures, culture times and identification characteristics of various intestinal flora used for detection of various intestinal flora are shown in table 4.
TABLE 4 culture Medium, culture temperature, culture time and identification characteristics
Figure BFW0000023323610000101
The results are shown in FIG. 2. After a mouse type 2 diabetes model is constructed by combining high-fat feed and streptozotocin, the intestinal flora of the mouse is disordered. The amount of lactobacillus and bifidobacterium in the intestinal tract of the Sc52 group mice gradually increased along with the prolongation of the gavage time, the intestinal tract of the mice basically recovered to the level of a blank control group at the 14 th week, and the amount of the lactobacillus and the bifidobacterium in the intestinal tract of the mice slightly decreased after the gavage administration is stopped. The quantity of enterobacteria, enterococcus and clostridium perfringens in intestinal tracts of mice in the Sc52 group gradually decreases along with the prolonging of treatment time, the level of the mice is basically recovered to the level of a blank control group at the 14 th week, and the bacterial population quantity has no obvious trend change after the gastric lavage administration is stopped. The results show that the lactobacillus plantarum Sc52 increases the number of two main probiotics, namely lactobacillus and bifidobacterium, forms a biological barrier together with other anaerobic flora, inhibits the colonization of pathogenic bacteria such as enterobacteria and enterococcus, regulates the balance of intestinal microorganisms and promotes the development of organisms to a healthy direction.

Claims (1)

1. Application of lactobacillus plantarum Sc52 in preparation of products; the preservation number of the lactobacillus plantarum (Lactobacillus plantarum) Sc52 is CGMCC NO. 11027;
the product functions as at least one of (b1) to (b9) as follows:
(b1) reducing blood sugar;
(b2) reducing blood glucose levels in serum;
(b3) reducing fasting blood glucose;
(b4) improving glucose intolerance;
(b5) treatment and/or prevention of diabetes;
(b6) reducing blood sugar content in blood serum of diabetic;
(b7) reducing fasting blood glucose value of diabetic;
(b8) increasing the insulin content in the serum of a diabetic patient;
(b9) improving glucose intolerance in diabetic patients;
the product is a medicine, and the diabetes is type 2 diabetes.
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