CN108624520A - Promote the lactobacillus plantarum strain GMNL-662 and combinations thereof of inostosis - Google Patents
Promote the lactobacillus plantarum strain GMNL-662 and combinations thereof of inostosis Download PDFInfo
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- CN108624520A CN108624520A CN201710157055.7A CN201710157055A CN108624520A CN 108624520 A CN108624520 A CN 108624520A CN 201710157055 A CN201710157055 A CN 201710157055A CN 108624520 A CN108624520 A CN 108624520A
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- lactobacillus plantarum
- inostosis
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Classifications
-
- 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
-
- 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
-
- 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
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
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- Bioinformatics & Cheminformatics (AREA)
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- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
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- Tropical Medicine & Parasitology (AREA)
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Abstract
The present invention discloses a kind of lactobacillus plantarum strain GMNL 662 and combinations thereof promoting inostosis.The lactobacillus plantarum strain GMNL 662 has the ability for improving osteogenesis gene expression, inhibiting the expression of erosion bone related genes and improving the relevant cytohormone TGF β expression of ostosis, therefore can delay bone-loss.
Description
Technical field
The invention relates to a kind of lactobacillus plantarum strain GMNL-662 and combinations thereof promoting inostosis, especially
It is about a kind of lactobacillus plantarum strain GMNL-662 of ability and combinations thereof for having and improving osteogenesis gene expression.
Background technology
Osteoporosis is a kind of systemic skeletal disease, and feature includes that bone amount is reduced, and the microstructure of bone tissue is deteriorated,
Cause bone fragile, fracture risk increases.
During remodeling, including the bon e formation effect of osteoblast and the bone resorption of Osteoclasts, it is common to tie up
The dynamic equilibrium of bone tissue is held, once bone information is more than bon e formation, i.e., can lead to bone loss, finally cause osteoporosis.One
As for, osteoporosis is divided into two class of osteoporosis and senile osteoporosis after menopause.Osteoporosis after menopause
Disease is common in women after menopause, since internal gynotermone amount is rapidly reduced after women menopause, Osteoclasts increased activity and absorb
Bone trabecula finally makes bone trabecula attenuate, wreck, number is reduced, discontinuous, decrease bone strength；And senile osteoporosis is
Osteoblast deterioration, calcium and vitamin D intake are insufficient, and intestinal absorption ability is deteriorated, and cause bone synthesis to reduce, cortex of bone
Thicker pine macropore, bone trabecula disappear, and bone strength obviously lowers.
The drug of prevention osteoporosis and fracture at present, according to its effect machine turn can be divided into bone against corrosion or anti-current lose class drug,
Rush makes bone or promotees ostosis class drug and mixed type three classes.Bone class drug against corrosion includes calcium agent, vitamin D, calcitonin, double phosphorus
Hydrochlorate, estrogenic agents, sex hormones, Osteoclasts ferment inhibitor, RANKL monoclonal antibodies etc., mixed type is current
Only strontium salt is a kind of.The drug of osteoporosis itself is prevented with some side effects.Clinical test results find to merge and use
Drug has no addition effect, can resist mutually instead, or increases the incidence or intensity of side effect, so various countries are each at present
Kind osteoporosis prevention is guided, and it is not recommended that two kinds of anti-loss agents are used in combination, or anti-loss agent is used in combination to make bone agent with rush.
Clinically be commonly used for the osteoporosis drug of old man and climacteric women, as Fosamax ingot, gram bone pine ingot, can bone
The Diphosphonates similar drug such as magnificent ingot does not pay attention to oral hygiene or is had tooth pulled out, dental implant surgical, serious jaw probably occurs if taking for a long time
Bones and joints necrosis；Recent research is also found, may cause the adverse reaction of atypia femoral fracture.
Although past has part document to point out that certain specific probiotics strains have the sclerotin for reducing ovariectomy rats and mice
The ability of loss, such as：L.reuteri ATCC PTA 6475；L.paracasei DSM13434；L.plantarum DSM
15312, DSM 15313 and B.longum etc. is to be tested in the form of viable bacteria, and be the discovery that by reducing inflammatory response
Mechanism delays the ability of bone-loss to reach, but they and without the ability that can make inostosis, belong to more passive
Therapeutic modality.
Therefore, it is necessary to a kind of lactobacillus plantarum strain GMNL-662 and combinations thereof promoting inostosis is provided, with solution
The problems of certainly in the prior art.
Invention content
The main purpose of the present invention is to provide it is a kind of promote inostosis lactobacillus plantarum strain GMNL-662 and its
Composition, the lactobacillus plantarum strain GMNL-662 (Lactobacillus Plantarum GMNL-662) can be by any
Form enters in digestive system, increases the expression of cytohormone TGF-β and osteocalcin (Osteocalcin), at the same can inhibit and
The problem of reducing the performance amount of erosion bone related genes (such as TRAP-5), therefore reaching improvement bone-loss.
To reach the foregoing purpose of the present invention, one embodiment of the invention provides a kind of plant breast bar promoting inostosis
Bacterium (Lactobacillus Plantarum) bacterial strain, the lactobacillus plantarum strain is lactobacillus plantarum strain GMNL-662, and
It is preserved in China typical culture collection center (CCTCC) with deposit number M 2016571.
In one embodiment of this invention, the lactobacillus plantarum strain GMNL-662 is viable bacteria bacterial strain or dead bacteria strain.
In one embodiment of this invention, the lactobacillus plantarum strain GMNL-662, which has, improves osteogenesis gene expression
Ability.
The osteogenesis gene is osteocalcin (Osteocalcin) gene.
To reach the foregoing purpose of the present invention, another embodiment of the present invention provides a kind of combination promoting inostosis
Object, it includes the lactobacillus plantarum strains as described above for promoting inostosis.
In one embodiment of this invention, the composition of the promotion inostosis is a medical composition, nutrition benefit
Fill product, a health food, a dietetic food or combinations thereof.
Description of the drawings
The performance amount bar chart of the cytohormone TGF-β of each group in Fig. 1 displays experiment 2.
The performance amount bar chart of the osteogenesis gene Osteocalcin of each group in Fig. 2 displays experiment 2.
The performance amount bar chart of the erosion bone related genes TRAP-5 of each group in Fig. 3 displays experiment 2.
Specific implementation mode
In order to allow the present invention above-mentioned and other purposes, feature, advantage can be clearer and more comprehensible, hereafter by spy lift the present invention compared with
Good embodiment, and coordinate institute's accompanying drawings, it is described in detail below.In addition, the singulative " one " that the present invention is previously mentioned, "one"
" described " includes several references, unless the context clearly determines otherwise.Numberical range (such as 10% to 11% A) is if without specific
Illustrate all to include upper limit value and lower limit value (i.e. 10%≤A≤11%)；If numberical range do not define lower limiting value (as be less than 0.2% B, or
0.2% B below), then it may be 0 (i.e. 0%≤B≤0.2%) all to refer to its lower limiting value.Above-mentioned term is to illustrate and understand
The present invention, rather than to limit the present invention.
One embodiment of the invention provides a kind of lactobacillus plantarum strain promoting inostosis, the lactobacillus plantarum bacterium
Strain is lactobacillus plantarum strain GMNL-662 (Lactobacillus Plantarum GMNL-662), and with deposit number M
2016571 are preserved in China typical culture collection center (CCTCC).
One embodiment of the invention provides a kind of composition promoting inostosis, and it includes plant breast bars as described above
Bacteria strain GMNL-662.Preferably, the composition for promoting inostosis can also a medical composition, a nutritional supplementation
The mode of product, a health food, a dietetic food or combinations thereof is presented, that is, the composition for promoting inostosis can root
According to validity or convenience different forms is designed to consider.In addition, it is described promote inostosis composition preferably with
Edible way enters digestive system, and then promotes the expression of osteogenesis gene, inhibits the performance of erosion bone gene and increase protection sclerotin
The expression of the cytohormone TGF-β of loss, to delay bone-loss.
The lactobacillus plantarum strain GMNL-662 in above-described embodiment is mainly obtained from human body intestinal canal screening separation
Wherein one plant of multiple separation strains.Utilize introduction (the SEQ ID NO of table 1:1 and SEQ ID NO:2) it is more to replicate to carry out PCR
A respective 16S rDNA segments of separation strains are then subject to sequencing, wherein one plant of 16S rDNA are obtained after the completion of sequencing
Following (the SEQ ID NO of gene order:3)；Then, as shown in table 2 below, the result after being compared by the websites NCBI is it is found that described point
16S rDNA sequences from strain are similar to the 16S rDNA sequences of lactobacillus plantarum (Lactobacillus Plantarum), phase
It is 99% or more like degree, therefore understands that this bacterial strain GMNL-662 really belongs to lactobacillus plantarum.
Table 1, primer sequences control
| Introduction | Sequence number | Sequence |
| PAF | SEQ ID NO:1 | AGA GTT TGA TCC TGG CTC AG |
| 536R | SEQ ID NO:2 | GTA TTA CCG CGG CTG CTG |
Table 2, strain sequence deck watch
| NCBI accession number | Bacterial strain scientific name | Similarity |
| KT236093.1 | Lactobacillus plantarum KLB 410 | 99% |
| KT962240.1 | Lactobacillus plantarum USIM03 | 99% |
| KT025848.1 | Lactobacillus plantarum KF | 99% |
| KR816164.1 | Lactobacillus plantarum KF9 | 99% |
Complete 16S rDNA sequences (the SEQ ID NO of the lactobacillus plantarum GMNL-662:3) as follows：
GCCGTTGGCGTCGGATACATGCATGTCGTACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTAC
ATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGAT
GCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCC
GCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCC
ACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGACGAAAGTCTGAT
GGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAA
CTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGGTAAACAC
The lactobacillus plantarum strain GMNL-662 Jin Hang Fermentation ferment is tested, the result such as table 3 is can get.
3, Fermentation ferment test results of table
There is the spy for promoting inostosis in order to verify the lactobacillus plantarum strain GMNL-662 provided by the present invention
Property, and the symptom for improving bone-loss can be reached, carry out following experiment.
Experiment 1：Bone tissue is analyzed
Bacterial strain：Lactobacillus plantarum (Lactobacillus plantarum；GMNL-662)
Bacterial strain processing：
(1) prepared by viable bacteria：From frozen pipe inoculated plant lactobacillus strain (Lactobacilli plantarum GMNL-662)
To 1ml MRS broth, and in 37 DEG C of aerobic lower stationary cultures 20 hours.Next day takes 15 μ l culture bacterium solutions overnight to 1.5ml
MRS broth (1% re-activation) then estimate bacterium number in aerobic lower stationary culture 20 hours with 37 DEG C with OD 600nm, and
Viable bacteria dosage is adjusted to 8 × 107Cfu/ml is spare.
(2) prepared by dead bacterium：From frozen pipe inoculated plant lactobacillus (Lactobacilli plantarum GMNL-662) to
1ml MRS broth, and in 37 DEG C of aerobic lower stationary cultures 20 hours.Next day takes 15 μ l culture bacterium solutions overnight to 1.5ml MRS
Broth (1% re-activation) then in 37 DEG C of aerobic lower stationary cultures 20 hours, then with OD 600nm estimates bacterium number, and by bacterium
Number is adjusted to 4.1 × 108Cfu/ml, and 121 DEG C of progress autoclave sterilization 15 minutes is spare.
Osteoporosis mouse model：
8 week old ICR female mices are bought by Le Sikesheng skills company, removal ovary operation is carried out when 9 week old.Mouse is being anaesthetized
Under, it is cut at the ovary of back both sides, carries out removal ovary operation；All groups start pipe for after surgery 4 days and feed administering experiment
Substance.Mouse is divided into sham-operation group (Control groups only open abdominal cavity, do not extract ovary)；And 4 groups of ovariectomized groups
(Ovariectomy；OVX).When mouse is sacrificed, ovary tissue is checked, confirm whether removal operation on ovary succeeds, operative failure
Animal, experimental result do not use.In the mouse of 4 groups of ovariectomized groups, it is solvent group (H to take 1 group2O groups), 1 group is positive control drug
(osteosporosis resistant medicament Alendronate).Alendronate is small with the suspension of deionized water compound concentration 0.25mg/ml
The every 10 g of weight of mouse administer 0.1ml, administer 4 times within one week.Remaining 2 component, which don't bother about, feeds GMNL-662 viable bacteria 0.2ml (viable bacteria concentrations
It is 8 × 107cfu/ml；Convert mouse daily dosage 1.6 × 107Cfu/mouse, human dose 4 × 109Cfu/60kg is grown up),
And (dead bacteria concentration is 4.1 × 10 to dead bacterium 0.2ml8cfu/ml；Convert mouse dose 8.2 × 107Cells/mouse, convert people
Body dosage：2×1010Cells/60kg is grown up), two groups respectively pipe is fed once, after continuous 28 days, by mouse anesthesia and with abdominal cavity daily
Venous blood collection sacrifice takes out femur for analysis.
Analysis method：
Right femur distal end backbone is with Microcomputerized tomography scanner (micro computed tomography；
SkyScan 1076, Kontizh, Belgium, resolution are 18 μm) shooting computed tomography figure, and to analyze software point
Analyse the ratio in bone trabecula region, the i.e. ratio (bone volume/tissue volume) of bone volume and tissue volume.Analysis
The region of (toward proximal part) 100, does not include cortex bone under the growth selection plate of position.The analysis of bone mineral density selects same zone
The bone mineral density without cortex bone is analyzed respectively in domain.The data for testing gained, with double tail analysis of variance (two-way
Analysis of variance), and T-test statistical analyses are carried out, all data are with average ± standard deviation (mean ± SD)
It indicates；Each group and OVX+H2O groups compare after through statistical analysis, indicate statistically to have that (* represents p ＜ to significant difference with distinct symbols
0.05；* represents p ＜ 0.01).The result of experiment 1 is see table 4 and table 5.
Table 4, bone trabecula region ratio (BV/TV)
As can be seen from Table 4, after mouse removal ovary, disease group (OVX+H is made2The ratio in bone trabecula region O), i.e. bone volume
With the ratio analysis of tissue volume, all substantially less than sham-operation group (Control), indicate osteoporosis zootype make disease at
Work(.Compare and feed this two groups of GMNL-662 viable bacterias and dead bacterium, it is possible to find BV/TV analyses, which are significantly higher than, makes disease group, indicates GMNL-
662 can slow down the degree of bone-loss after ovariectomy really.Alendronate is positive control group, and also tool protection ovary is plucked
Except the degree of rear bone-loss；And feed GMNL-662 bacterial strains two groups are managed, protecting effect is even slightly better than osteosporosis resistant medicament
Alendronate。
Table 5, femur bone mineral density (being free of cortex bone, BMD)
As can be seen from Table 5, disease group (OVX+H is made2O right femur bone mineral density) is all substantially less than group of doing evil through another person
(Control)；And the group of GMNL-662 viable bacterias and dead bacterium is fed, bone density, which is substantially better than, makes disease group (OVX+H2O), table
Show that pipe feeds two groups of GMNL-662 degree that can slow down bone-loss after ovariectomy.
Experiment 2：GMNL-662 is to osteogenesis gene, cytohormone and the influence for losing bone gene
Extract shin bone RNA：Mouse left femur is removed after experiment, shin bone is cut into small pieces with scissors, and is added suitable
The liquid nitrogen of amount makes bone be ground rapidly；0.5ml is added in the bone meal groundReagent carries out RNA extractions,
Be added 0.1ml chloroforms (chloroform) spin upside down 15 times, be placed in room temperature reaction after five minutes centrifugation and by upper layer extract to
In new microcentrifugal tube eppendorf, and 0.25ml isopropanols are added, are centrifuged after ten minutes being placed at room temperature for；It removes
The sediment after the cleaning centrifugation of 75% ethyl alcohol of 0.5ml is used in combination in supernatant；20-50 is added after drying up the sediment after above-mentioned centrifugation
DEPC water (diethylpyrocarbonate treated water) back dissolving of μ l measures RNA concentration.
RNA inverts cDNA：It takes 1-5 μ g RNA and mends without RNA enzyme water (RNase-free water) to 10 μ l；In addition then add
Enter 10 × Ramdon primer (2 μ l), 10mM dNTP (1 μ l), 65 DEG C 5 minutes, on ice 2-3 minutes；It is finished the first stage
Afterwards, 5 × RT buffer (4 μ l) are added, 0.1M DTT (1 μ l), RNase inhibitor (RNase inhibitor,
Invitrogen, RNaseOUTTM, 1 μ l), RT ferment ((Invitrogen,1 μ l), RT points of Mix
Clock is placed in 50 DEG C 15 minutes in 60 minutes and 70 DEG C, carries out the effect of reverse transcription ferment.
Shin bone cDNA carries out real-time PCR analyses：Take 1 μ l shin bones cDNA that 4 μ L, 1 μM of F+R primers are added
The 2x Rotor-Gene SYBR Green PCR Master Mix of (forwards/reverse primer sequences are as follows), 5 μ L
(Qiagen, Cat.204076) is put into Q-PCR machines and is reacted, and TGF-β and RANKL relative expression quantities are deduction itself
Obtained by GAPDH.
Table 5, primer sequences
Analysis method：The data for testing gained, with double tail analysiss of variance (two-way analysis of
Variance), and T-test is carried out.Statistical analysis each group is and OVX+H2O groups are compared, and * represents p ＜ 0.05；* represents p ＜
0.01。
As shown in Figure 1, GMNL-662 viable bacterias and dead bacterial type state can increase the cytohormone TGF-β of protection bone-loss
Performance.After mouse removal ovary compared with sham-operation group, ostosis relevant cell hormone gene TGF-β performance amount is substantially reduced；
The TGF-β performance amount of feeding GMNL-662 groups (the dead bacterium of GMNL-662 and viable bacteria) and ovariectomy group (OVX+H2O) comparing has
The situation obviously increased, this result indicates that GMNL-662 has effects that promote TGF-β expression, and thereby delays bone-loss.
Then, as shown in Fig. 2, giving the GMNL-662 groups of (viable bacteria and dead bacterium), osteogenesis gene after removal ovary
The performance amount of Osteocalcin, which is above, makes disease group (OVX+H2O), indicate that GMNL-662 has the work(for promoting osteogenesis gene expression
Effect, GMNL-662 viable bacterias and dead bacterial type state can improve the performance of osteogenesis gene Osteocalcin, and thereby delay sclerotin stream
It loses.
Please continue to refer to Fig. 3, it can be clearly seen that, after mouse removes ovary, make disease group (OVX+H2) and sham-operation group O
(Control) it compares, erosion bone related genes TRAP-5 performance amounts have increased situation；And give pipe feed GMNL-662 viable bacterias and
The group of dead bacterium, the performance amount of erosion bone gene TRAP-5, which is significantly lower than, makes disease group (OVX+H2O), indicate that GMNL-662 has
The effect of inhibiting erosion bone gene expression, and thereby delay bone-loss.
In summary result, it may be determined that lactobacillus plantarum GMNL-662 provided by the present invention, whether viable bacteria or heat
Bacterial type state is killed, in the bone tissue analysis (ratio in bone trabecula region, BV/TV) of zoopery, femur bone mineral density (BMD)
The bone-loss phenomenon of ovariectomy mouse can significantly be slowed down；And find that GMNL-662 has promotion osteogenesis gene
(Osteocalcin), inhibit the expression of erosion bone gene (TRAP-5) and increase the cytohormone TGF-β expression of protection bone-loss
Ability, and then achieve the purpose that inhibit bone-loss.
In addition, experimental result is it has also been found that the protection bone-loss effect of GMNL-653 is better than Forteo
Alendronate.And at present clinically it has been found that Alendronate has many side effects, including heart disease, obstinate pain
Disease, jawbone necrosis, fracture, cancer of the esophagus etc.；Therefore, safe and without side-effects lactobacillus plantarum strain GMNL-662 is applied to prolong
Should should be able to be a kind of better choice for the bone loss disorders of women after the following prevention and improvement menopause on slow bone-loss.
The present invention is described by above-mentioned related embodiment, however above-described embodiment is only the example for implementing the present invention.
It must be noted that, it has been disclosed that embodiment be not limiting as the scope of the present invention.On the contrary, being contained in the spirit of claims
And range modification and impartial setting be included in the scope of the present invention.
SEQUENCE LISTING
<110>Jingyue Biological Science and Technology Co., Ltd.
<120>Promote the lactobacillus plantarum strain GMNL-662 and combinations thereof of inostosis
<130> TP161062-TW
<160> 11
<170> PatentIn version 3.5
<210> 1
<211> 20
<212> DNA
<213>Artificial sequence
<220>
<223>PAF introductions
<400> 1
agagtttgat cctggctcag 20
<210> 2
<211> 18
<212> DNA
<213>Artificial sequence
<220>
<223>536R introductions
<400> 2
gtattaccgc ggctgctg 18
<210> 3
<211> 525
<212> DNA
<213>Artificial sequence
<220>
<223>GMNL-662 16S rDNA sequence introductions
<400> 3
gccgttggcg tcggatacat gcatgtcgta cgaactctgg tattgattgg tgcttgcatc 60
atgatttaca tttgagtgag tggcgaactg gtgagtaaca cgtgggaaac ctgcccagaa 120
gcgggggata acacctggaa acagatgcta ataccgcata acaacttgga ccgcatggtc 180
cgagcttgaa agatggcttc ggctatcact tttggatggt cccgcggcgt attagctaga 240
tggtggggta acggctcacc atggcaatga tacgtagccg acctgagagg gtaatcggcc 300
acattgggac tgagacacgg cccaaactcc tacgggaggc agcagtaggg aatcttccac 360
aatgacgaaa gtctgatgga gcaacgccgc gtgagtgaag aagggtttcg gctcgtaaaa 420
ctctgttgtt aaagaagaac atatctgaga gtaactgttc aggtattgac ggtatttaac 480
cagaaagcca cggctaacta cgtgccagca gccgcgggta aacac 525
<210> 4
<211> 20
<212> DNA
<213>Artificial sequence
<220>
<223>TGF beta forward direction introductions
<400> 4
gagtaacgct ttccggagtc 20
<210> 5
<211> 20
<212> DNA
<213>Artificial sequence
<220>
<223>The reversed introductions of TGF beta
<400> 5
acagtcacca gcatctcagc 20
<210> 6
<211> 25
<212> DNA
<213>Artificial sequence
<220>
<223>Osteocalcin forward direction introductions
<400> 6
acggtatcac tatttaggac ctgtg 25
<210> 7
<211> 24
<212> DNA
<213>Artificial sequence
<220>
<223>The reversed introductions of Osteocalcin
<400> 7
actttatttt ggagctgctg tgac 24
<210> 8
<211> 20
<212> DNA
<213>Artificial sequence
<220>
<223>TRAP-5 forward direction introductions
<400> 8
gacgatgggc gctgacttca 20
<210> 9
<211> 20
<212> DNA
<213>Artificial sequence
<220>
<223>The reversed introductions of TRAP-5
<400> 9
gcgcttggag atcttagagt 20
<210> 10
<211> 20
<212> DNA
<213>Artificial sequence
<220>
<223>GAPDH forward direction introductions
<400> 10
gcacagtcaa ggccgagaat 20
<210> 11
<211> 20
<212> DNA
<213>Artificial sequence
<220>
<223>The reversed introductions of GAPDH
<400> 11
gccttctcca tggtggtgaa 20
Claims (9)
1. a kind of lactobacillus plantarum strain promoting inostosis, which is characterized in that the lactobacillus plantarum strain is plant breast
Bacillus strain GMNL-662, and the lactobacillus plantarum strain GMNL-662 is preserved in China with deposit number M 2016571
Type Tissue Collection.
2. promoting the lactobacillus plantarum strain of inostosis as described in claim 1, it is characterised in that：The lactobacillus plantarum
Bacterial strain GMNL-662 is viable bacteria bacterial strain or dead bacteria strain.
3. promoting the lactobacillus plantarum strain of inostosis as described in claim 1, it is characterised in that：The lactobacillus plantarum
Bacterial strain GMNL-662 has the ability for improving osteogenesis gene expression.
4. promoting the lactobacillus plantarum strain of inostosis as claimed in claim 3, it is characterised in that：The osteogenesis gene is
Osteocalcin gene.
5. a kind of composition promoting inostosis, which is characterized in that the composition includes as described in claim 1 promotes
The lactobacillus plantarum strain of inostosis.
6. promoting the composition of inostosis as claimed in claim 5, it is characterised in that：The combination for promoting inostosis
Object is a medical composition, a nutriment, a health food, a dietetic food or combinations thereof.
7. promoting the composition of inostosis as claimed in claim 5, it is characterised in that：The lactobacillus plantarum strain
GMNL-662 is viable bacteria bacterial strain or dead bacteria strain.
8. promoting the composition of inostosis as claimed in claim 5, it is characterised in that：The lactobacillus plantarum strain
GMNL-662 has the ability for improving osteogenesis gene expression.
9. promoting the composition of inostosis as claimed in claim 8, it is characterised in that：The osteogenesis gene is osteocalcin base
Cause.
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| WO2022223005A1 (en) * | 2019-06-25 | 2022-10-27 | 生合生物科技股份有限公司 | Method for promoting bone healing using lactiplantibacillus plantarum twk10 |
| CN115806898A (en) * | 2022-08-10 | 2023-03-17 | 重庆第二师范学院 | Lactobacillus plantarum KSFY04 and application thereof |
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