CN106619742A - Lactic acid bacteria composition used for treating diabetes, and preparation method thereof - Google Patents
Lactic acid bacteria composition used for treating diabetes, and preparation method thereof Download PDFInfo
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- CN106619742A CN106619742A CN201610980275.5A CN201610980275A CN106619742A CN 106619742 A CN106619742 A CN 106619742A CN 201610980275 A CN201610980275 A CN 201610980275A CN 106619742 A CN106619742 A CN 106619742A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
Abstract
The invention discloses a lactic acid bacteria composition used for treating diabetes. The lactic acid bacteria composition used for treating diabetes comprises a lactic acid bacteria combination nuclear; the lactic acid bacteria combination nuclear is coated with a coating layer; the coating layer comprises, by weight, 30 to 50 parts of acarbose, 30 to 50 parts of a protein substance, and 5 to 10 parts of nitrocellulose; the lactic acid bacteria combination nuclear is composed of, by weight, 20 to 30 parts of bifidobacterium longum, 10 to 20 parts of lactobacillus salivarius, 20 to 30 parts of lactococcus lactis, 10 to 20 parts of lactobacillus rhamnosus, and 10 to 20 parts of lactobacillus acidophilus. The invention also discloses a preparation method of the lactic acid bacteria composition. The lactic acid bacteria composition can be used for treating diabetes with obvious effect, and no toxic or side effect is caused.
Description
Technical field
The present invention relates to probio, can significantly treat diabetes and without any side effects control more particularly, to a kind of
Treat lactic bacteria composition of diabetes and preparation method thereof.
Background technology
Diabetes are one group of metabolic diseases being characterized with hyperglycaemia.Hyperglycaemia be then due to defect of insulin secretion or
Its biological agent is damaged, or both have concurrently and cause.Long-standing hyperglycaemia during diabetes, cause various tissues, particularly eye,
Kidney, heart, blood vessel, chronic lesion, the dysfunction of nerve.With the aging of world population, diabetes have become one kind
Common disease, frequently-occurring disease, in industrially developed country, its incidence of disease is in rising trend.According to statistics, the whole world there are about 1.5 hundred million patient of diabetes
Person.The death toll caused by diabetic complication has been the rank rear the 3rd after cardiovascular and cerebrovascular disease, cancer in developed country,
The great attention of countries in the world is caused, pathogenesis, medical treatment one after another to diabetes etc. are studied.Current medical circle
It is taken be include dietetic treatment, chemoprophylaxis and treatment with and other complementary therapies comprehensive prevention and remedy measures.
Although Western medicine emerges in an endless stream, but can not tackle the problem at its root, and Western medicine prevention and treatment have certain side effect.
The content of the invention
To overcome the shortcoming of prior art, present invention aim at providing one kind can significantly treat diabetes and without any
Lactic bacteria composition for the treatment of diabetes of toxic and side effect and preparation method thereof.
The present invention is by following technical measures realization, a kind of lactic bacteria composition for treating diabetes, including lactic acid bacteria
Combination core, the lactic acid bacteria combination core is wrapped with coated layer；The coated layer includes the composition of parts by weight：Acarbose 30
～50, protein substance 30～50, celluloid 5～10；The lactic acid bacteria combination core is made up of the bacterial strain of following parts by weight：
Bifidobacterium longum 20～30, Lactobacillus salivarius 10～20, Lactococcus lactis 20～30, Lactobacillus rhamnosus 10～20, acidophilus breast
Bacillus 10～20.
Used as a kind of preferred embodiment, the content of bifidobacterium longum is 10 in the lactic bacteria composition6～109Cuf/g, saliva
The content of liquid lactobacillus is 106～108Cuf/g, the content of Lactococcus lactis are 106～108Cuf/g, Lactobacillus rhamnosus contain
Measure as 106～107Cuf/g, the content of lactobacillus acidophilus are 106～107cuf/g。
As a kind of preferred embodiment, bifidobacterium longum in the lactic acid bacteria combination core, Lactobacillus salivarius, Lactococcus lactis,
, respectively through resistance to Pig cholate, acidproof screening, the Pig cholate and pH for being resistant to 0.3% be for Lactobacillus rhamnosus and lactobacillus acidophilus
2.5 sour environment.
Used as a kind of preferred embodiment, the protein substance is casein and lactoferrin layer, wherein casein and newborn iron egg
White weight ratio is 1-5：1-5.
Also a kind of preparation method of the lactic bacteria composition for treating diabetes of the present invention, comprises the steps：
(1) by bifidobacterium longum, Lactobacillus salivarius, Lactococcus lactis, Lactobacillus rhamnosus, lactobacillus acidophilus respectively with
The inoculum concentration of 1ml/100ml is seeded in culture medium, at 35 DEG C～45 DEG C, cultivates 12～24h, obtains the lactobacillus bacterium for activating
Kind；
(2) by five kinds of bacterial classifications of activation with parts by weight：Bifidobacterium longum 20～30, Lactobacillus salivarius 10～20, lactic acid
After galactococcus 20～30, Lactobacillus rhamnosus 10～20, lactobacillus acidophilus 10～20 mix, then by mixed bacteria with 1ml/
The inoculum concentration of 100ml is seeded in nutrient solution, at 35 DEG C～45 DEG C, after 10～12h of culture, and centrifuging and taking precipitation, 4 DEG C～10
It is incubated after 6～10h at DEG C, homogeneous obtains combination strain；
(3) by parts by weight：Acarbose 30～50, protein substance 30～50, celluloid 5～10 add water fully
Coating buffer is mixed into, combination strain addition coating buffer is mixed evenly；
(4) liquid being mixed evenly is added drop-wise in calcium chloride solution and is solidified, vacuum freeze-drying 12h obtains lactic acid bacteria after filtration
Composition.
Used as a kind of preferred embodiment, the content of bifidobacterium longum is 10 in the lactic bacteria composition6～109Cuf/g, saliva
The content of liquid lactobacillus is 106～108Cuf/g, the content of Lactococcus lactis are 106～108Cuf/g, Lactobacillus rhamnosus contain
Measure as 106～107Cuf/g, the content of lactobacillus acidophilus are 106～107cuf/g。
As a kind of preferred embodiment, the bifidobacterium longum, Lactobacillus salivarius, Lactococcus lactis, Lactobacillus rhamnosus and
Lactobacillus acidophilus is resistant to 0.3% Pig cholate and acid that pH is 2.5 before activation respectively through resistance to Pig cholate, acidproof screening
Property environment.
Used as a kind of preferred embodiment, the protein substance is casein and lactoferrin layer, wherein casein and newborn iron egg
White weight ratio is 1-5：1-5.
The present invention has the effect of significantly treatment diabetes, and because being prebiotic becteriums product, does not have any poison to human body
Side effect.
Specific embodiment
The present invention is described in further detail with reference to embodiment.
Embodiment 1
(1) actication of culture
Filter out resistance to 0.3% Pig cholate, the bifidobacterium longum of the sour environment that resistance to PH is 2.5, Lactobacillus salivarius, lactic acid breast
Coccus, Lactobacillus rhamnosus and lactobacillus acidophilus, filter out five kinds of bacterium are seeded to respectively with the inoculum concentration of 1ml/100ml
In MRS culture mediums, at 45 DEG C, 12h is cultivated, obtain the lactobacillus strain for activating；
Wherein, the formula of MRS culture mediums is：10g peptones, 8g beef extract powders, 20g Portugals are added in 1000mL distilled water
Grape sugar, 4g yeast extracts, the water manganese sulfates of 0.05g tetra-, 1mL Tween 80s, 2g dipotassium hydrogen phosphates, 5g sodium acetates, 0.2g magnesium sulfate, 2g
Dibasic ammonium citrate；
(2) combination strain is prepared
By four kinds of bacterial classifications of activation with parts by weight：Bifidobacterium longum 20g, Lactobacillus salivarius 10g, Lactococcus lactis 20g,
After Lactobacillus rhamnosus 10g, lactobacillus acidophilus 10g mixing, then mixed bacteria is seeded to into culture with the inoculum concentration of 1ml/100ml
In liquid, at 45 DEG C, after culture 10h, with 6000r/min, at 5 DEG C, 15min is centrifuged, taking precipitate is incubated 6h at 4 DEG C
Afterwards, homogeneous, obtains combination strain；
(3) coating combination strain is prepared
By following weight：Acarbose 30g, casein 15g, lactoferrin 15g, celluloid 5g add water 1000ml
Coating buffer is sufficiently mixed into, combination strain addition coating buffer is mixed evenly；
(4) lactic bacteria composition is prepared
The liquid being mixed evenly is added drop-wise in calcium chloride solution and is solidified, vacuum freeze-drying 12h obtains lactic acid bacteria group after filtration
Compound.In lactic bacteria composition the content of bifidobacterium longum be 106～109cuf/g, Lactobacillus salivarius content be 106～
108cuf/g, the content of Lactococcus lactis are 106～108Cuf/g, the content of Lactobacillus rhamnosus are 106～107It is cuf/g, thermophilic
The content of Lactobacillus lactis is 106～107cuf/g。
Embodiment 2
Filter out resistance to 0.3% Pig cholate, the bifidobacterium longum of the sour environment that resistance to PH is 2.5, Lactobacillus salivarius, lactic acid breast
Coccus, Lactobacillus rhamnosus and lactobacillus acidophilus, filter out five kinds of bacterium are seeded to respectively with the inoculum concentration of 1ml/100ml
In MRS culture mediums, at 35 DEG C, 24h is cultivated, obtain the lactobacillus strain for activating；
Wherein, the formula of MRS culture mediums is：10g peptones, 8g beef extract powders, 20g Portugals are added in 1000mL distilled water
Grape sugar, 5g yeast extracts, the water manganese sulfates of 0.06g tetra-, 1mL Tween 80s, 2g dipotassium hydrogen phosphates, 5g sodium acetates, 0.2g magnesium sulfate, 2g
Dibasic ammonium citrate；
(2) combination strain is prepared
By four kinds of bacterial classifications of activation with parts by weight：Bifidobacterium longum 30g, Lactobacillus salivarius 20g, Lactococcus lactis 30g,
After Lactobacillus rhamnosus 20g, lactobacillus acidophilus 20g mixing, then mixed bacteria is seeded to into culture with the inoculum concentration of 1ml/100ml
In liquid, at 35 DEG C, after culture 12h, with 4500r/min, at 10 DEG C, 20min is centrifuged, taking precipitate is incubated at 10 DEG C
After 10h, homogeneous obtains combination strain；
(3) coating combination strain is prepared
By following weight：Acarbose 50g, casein 35g, lactoferrin 35g, celluloid 10g add water
1000ml is sufficiently mixed into coating buffer, and combination strain addition coating buffer is mixed evenly；
(4) lactic bacteria composition is prepared
The liquid being mixed evenly is added drop-wise in calcium chloride solution and is solidified, vacuum freeze-drying 12h obtains lactic acid bacteria group after filtration
Compound.In lactic bacteria composition the content of bifidobacterium longum be 106～109cuf/g, Lactobacillus salivarius content be 106～
108cuf/g, the content of Lactococcus lactis are 106～108Cuf/g, the content of Lactobacillus rhamnosus are 106～107It is cuf/g, thermophilic
The content of Lactobacillus lactis is 106～107cuf/g。
Embodiment 3
Filter out resistance to 0.3% Pig cholate, the bifidobacterium longum of the sour environment that resistance to PH is 2.5, Lactobacillus salivarius, lactic acid breast
Coccus, Lactobacillus rhamnosus and lactobacillus acidophilus, filter out five kinds of bacterium are seeded to respectively with the inoculum concentration of 1ml/100ml
In MRS culture mediums, at 42 DEG C, 13h is cultivated, obtain the lactobacillus strain for activating；
Wherein, the formula of MRS culture mediums is：10g peptones, 8g beef extract powders, 20g Portugals are added in 1000mL distilled water
Grape sugar, 4g yeast extracts, the water manganese sulfates of 0.05g tetra-, 1mL Tween 80s, 2g dipotassium hydrogen phosphates, 5g sodium acetates, 0.2g magnesium sulfate, 2g
Dibasic ammonium citrate；
(2) combination strain is prepared
By four kinds of bacterial classifications of activation with parts by weight：Bifidobacterium longum 35g, Lactobacillus salivarius 15g, Lactococcus lactis 23g,
After Lactobacillus rhamnosus 15g, lactobacillus acidophilus 15g mixing, then mixed bacteria is seeded to into culture with the inoculum concentration of 1ml/100ml
In liquid, at 42 DEG C, after culture 11h, with 7000r/min, at 5 DEG C, 10min is centrifuged, taking precipitate is incubated 10h at 4 DEG C
Afterwards, homogeneous, obtains combination strain；
(3) coating combination strain is prepared
By parts by weight：Acarbose 35g, casein 20g, lactoferrin 20g, celluloid 6g add water 1000ml
Coating buffer is sufficiently mixed into, combination strain addition coating buffer is mixed evenly；
(4) lactic bacteria composition freeze-dried powder is prepared
The liquid being mixed evenly is added drop-wise in calcium chloride solution and is solidified, vacuum freeze-drying 12h obtains lactic acid bacteria group after filtration
Compound freeze-dried powder.The content of bifidobacterium longum is 106～109cuf/g, Lactobacillus salivarius in lactic bacteria composition freeze-dried powder
Content be 106～108cuf/g, Lactococcus lactis content be 106～108Cuf/g, the content of Lactobacillus rhamnosus are 106~
107Cuf/g, the content of lactobacillus acidophilus are 106～107cuf/g。
It is used to further illustrate the present invention below by clinical testing.
The clinical report of 100 patients with NIDDM of this lactic bacteria composition freeze-dried powder prevention and treatment is as follows.
1st, clinical observation
Lactic bacteria composition freeze-dried powder (preparing according to the method for embodiment 1) of the present invention is to treating patients with NIDDM 200
Example, the wherein male sex 100, women 100.
2nd, medicament selection
Treatment group's anthology invention lactic bacteria composition freeze-dried powder (is prepared) according to the method for embodiment 1, course for the treatment of (month
For a course for the treatment of), 1.5g/ bags,
A day three times, every time one bag, boiling water is taken after mixing it with water.
3rd, diagnostic criteria
Jing doctors trained in Western medicine are diagnosed, and meet ADA (ADA) diabetes diagnostic criterion in 2010, wherein man 100,
Female 100,45～72 years old age, the course of disease, 2～10 years.Prevention and treatment are observed before all patient's prevention and treatment after one month
The clinical symptoms such as soreness and weakness of waist and knees afterwards, fatigue and weak, dry mouth and throat, enuresis nocturna improve situation.Before prevention and treatment, after prevention and treatment
Fasting blood-glucose (FPG), glycosylated hemoglobin (HbA1c), FPI (FINS).
4th, criterion of therapeutical effect
It is effective：Clinical symptoms improve obvious, and soreness and weakness of waist and knees, fatigue and weak, dry mouth and throat, night pollakiuria etc. disappear or basic
Disappear, blood sugar level fluctuates in normal range (NR), blood pressure≤128/80mmHg, 24h Urine proteins ＜ 30mg, blood urea nitrogen and creatinine
It is down to normal level.
Effectively：Clinical symptom relief, soreness and weakness of waist and knees, fatigue and weak, dry mouth and throat, night pollakiuria etc. be improved significantly,
Fasting blood-glucose control is preferable, and fluctuation of blood pressure has decline in 140/90mmHg or so, 24h Urine proteins and blood urea nitrogen creatinine, but
Do not take a turn for the better completely.
It is invalid：Clinical symptoms improve substantially, and, without declining or having decline but undesirable, 24h Urine proteins are determined for blood pressure and blood sugar
Amount and blood urea nitrogen creatinine do not decline, or decline not up to effective standard.
5th, treatment results
Curative effect
| Treatment number | It is effective | Effectively | It is invalid | Total effective rate |
| 200 | 124 | 68 | 8 | 96% |
FBG, HbAlc and FINS compare before and after prevention and treatment
| Observation index | Before treatment | After treatment |
| FBG | 9.66±2.52 | 5.37±1.58 |
| HBALC | 7.64±2.37 | 6.31±1.13 |
| FINS | 17.24±4.11 | 10.02±1.35 |
6th, conclusion
As a result show, patient FBG, HBALC, FINS relatively prevent and treat front lower Jing after the prevention and treatment of 1 course for the treatment of
There is pole significant difference (P ＜ 0.01) before drop, with prevention and treatment, it is seen that medicine of the present invention can significantly reduce blood sugar, saccharification blood
The level of Lactoferrin and blood insulin, improves diabetic's insulin resistance.Jing after the prevention and treatment of 1 course for the treatment of, II type
Diabetic's clinical symptoms significantly mitigate, and the symptom such as soreness and weakness of waist and knees, fatigue and weak, dry mouth and throat, night pollakiuria disappears.
It is effective to have 124 in 200 patients with NIDDM, account for 62%；Effectively there are 68, account for 34%；It is invalid to have 8,4% is accounted for, always
Efficient is 96%.Over the course for the treatment of, 200 patients with NIDDM have no bad reaction.
It is more than that lactic bacteria composition to present invention treatment diabetes and preparation method thereof is set forth, is used to help
Understand the present invention, but embodiments of the present invention and be not restricted to the described embodiments, it is any without departing from institute under the principle of the invention
The change of work, modification, replacement, combination, simplify, should be equivalent substitute mode, be included in protection scope of the present invention it
It is interior.
Claims (8)
1. a kind of lactic bacteria composition for treating diabetes, it is characterised in that：Core, the lactic acid bacteria combination are combined including lactic acid bacteria
Core is wrapped with coated layer；The coated layer includes the composition of parts by weight：Acarbose 30～50, protein substance 30～50,
Celluloid 5～10；The lactic acid bacteria combination core is made up of the bacterial strain of following parts by weight：Bifidobacterium longum 20～30, saliva
Liquid lactobacillus 10～20, Lactococcus lactis 20～30, Lactobacillus rhamnosus 10～20, lactobacillus acidophilus 10～20.
2. lactic bacteria composition according to claim 1, it is characterised in that：Bifidobacterium longum in the lactic bacteria composition
Content be 106～109Cuf/g, the content of Lactobacillus salivarius are 106～108Cuf/g, the content of Lactococcus lactis are 106~
108Cuf/g, the content of Lactobacillus rhamnosus are 106～107Cuf/g, the content of lactobacillus acidophilus are 106～107cuf/g。
3. lactic bacteria composition according to claim 1, it is characterised in that：Long bifid bar in the lactic acid bacteria combination core
Bacterium, Lactobacillus salivarius, Lactococcus lactis, Lactobacillus rhamnosus and lactobacillus acidophilus respectively through resistance to Pig cholate, acidproof screening,
The Pig cholate of ability 0.3% and the sour environment that pH is 2.5.
4. lactic bacteria composition according to claim 1, it is characterised in that：The protein substance is casein and newborn iron egg
The weight ratio of white, wherein casein and lactoferrin is 1-5：1-5.
5. a kind of preparation method of the lactic bacteria composition for treating diabetes, it is characterised in that comprise the steps：
(1) by bifidobacterium longum, Lactobacillus salivarius, Lactococcus lactis, Lactobacillus rhamnosus, lactobacillus acidophilus respectively with 1ml/
The inoculum concentration of 100ml is seeded in culture medium, at 35 DEG C～45 DEG C, cultivates 12～24h, obtains the lactobacillus strain for activating；
(2) by five kinds of bacterial classifications of activation with parts by weight：Bifidobacterium longum 20～30, Lactobacillus salivarius 10～20, Lactococcus lactis
After bacterium 20～30, Lactobacillus rhamnosus 10～20, lactobacillus acidophilus 10～20 mix, then by mixed bacteria with 1ml/100ml's
Inoculum concentration is seeded in nutrient solution, and at 35 DEG C～45 DEG C, after 10～12h of culture, centrifuging and taking precipitation is protected at 4 DEG C～10 DEG C
After 6～10h of temperature, homogeneous obtains combination strain；
(3) by parts by weight：Acarbose 30～50, protein substance 30～50, celluloid 5～10 add water and are sufficiently mixed
Into coating buffer, combination strain addition coating buffer is mixed evenly；
(4) liquid being mixed evenly is added drop-wise in calcium chloride solution and is solidified, vacuum freeze-drying 12h obtains lactic acid bacteria combination after filtration
Thing.
6. preparation method according to claim 5, it is characterised in that：Bifidobacterium longum contains in the lactic bacteria composition
Measure as 106～109Cuf/g, the content of Lactobacillus salivarius are 106～108Cuf/g, the content of Lactococcus lactis are 106~
108Cuf/g, the content of Lactobacillus rhamnosus are 106～107Cuf/g, the content of lactobacillus acidophilus are 106～107cuf/g。
7. preparation method according to claim 5, it is characterised in that：The bifidobacterium longum, Lactobacillus salivarius, lactic acid breast
Coccus, Lactobacillus rhamnosus and lactobacillus acidophilus respectively through resistance to Pig cholate, acidproof screening, are resistant to 0.3% before activation
Pig cholate and pH are 2.5 sour environment.
8. preparation method according to claim 5, it is characterised in that：The protein substance is casein and lactoferrin
Layer, wherein the weight ratio of casein and lactoferrin are 1-5：1-5.
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| CN113583903A (en) * | 2021-07-23 | 2021-11-02 | 微康益生菌(苏州)股份有限公司 | Microbial composition for preventing or treating type II diabetes and preparation method and application thereof |
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| 孙欣: "吸附与包埋技术固定化乳酸菌及其性质研究", 《万方数据》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112075638A (en) * | 2019-06-14 | 2020-12-15 | 丰华生物科技股份有限公司 | Food compositions and pharmaceutical compositions comprising a hypoglycemic lactic acid bacterial strain |
| CN113583903A (en) * | 2021-07-23 | 2021-11-02 | 微康益生菌(苏州)股份有限公司 | Microbial composition for preventing or treating type II diabetes and preparation method and application thereof |
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Application publication date: 20170510 |