CN109715197A - Collagenic hydrolysate is used to improve the purposes of endurance performance and stimulation lipid catabolism - Google Patents
Collagenic hydrolysate is used to improve the purposes of endurance performance and stimulation lipid catabolism Download PDFInfo
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Abstract
It is used to improve the purposes of endurance performance by increasing mitochondria activity the present invention relates to collagenic hydrolysate.It is used to stimulate lipid catabolism by increasing mitochondria activity the invention further relates to collagenic hydrolysate, the purposes especially lost weight.
Description
The present invention relates to the purposes that collagenic hydrolysate is used to improve endurance performance.
In addition, being used to stimulate lipid catabolism the present invention relates to collagenic hydrolysate, especially for reducing weight
Purposes.
Collagenic hydrolysate, especially by the starting material of the animal origin comprising collagen enzymatic hydrolysis generate, be by
Peptide mixer composition, the molecular weight of the peptide depends on starting material and working condition and has dividing for certain size range
Cloth.Collagenic hydrolysate as nutritional supplement purposes it is known to the public have the comparable time, especially for prevent and/
Or treatment bone, joint or connective tissue disease, especially because having been proven that collagen peptide for the body in these organization types
The synthesis that body has extracellular matrix by oneself has stimulation (see, for example, Bello et al., Curr.Med.Res.Opin.2006
(22)2221-2232)。
It has now surprisingly been found that, collagenic hydrolysate also causes the active enhancing of humans and animals cell Mitochondria, that is, draws
Play the increase of the mitochondria number of each cell and/or the increase of individual mitochondria.
From this discovery it is found that collagenic hydrolysate of the invention can be used in a manner of targeting, increase for passing through
Mitochondria activity in the myocyte of humans and animals and improve endurance performance and stimulation lipid catabolism in humans and animals.
The endurance of human or animal's body shows and is used for ATP (atriphos) required for muscle systems supply for a long time
The ability of the aerobic metabolism of the energy of form is related.One key factor of aerobic capacity is the absorption of oxygen, again by three
A factor determines: utilizing by the oxygen supply of lung, by the oxygen transport of cardiovascular system and the oxygen in myocyte.Although maximum
Oxygen-supplying amount is substantially predefined by the anatomic environment (total surface area of alveolar) of individual, but can be by trained and other
Measure improves oxygen transport and oxygen utilizes, wherein the last one described step has typically represented conclusive limiting factor.Therefore,
Endurance, which shows, relies basically on (each myocyte's, or in entire muscle systems) mitochondria number, in respiratory chain
Consumption oxygen and the reaction for generating ATP occur in mitochondria.
In general, the increase of mitochondria activity means that the metabolic rate of body improves, and have per unit time a greater amount of
Nutrient be metabolized for energy regenerating.However, the activity of higher catabolism necessarily causes (if nutrition supply is kept
It is constant) the more havoc of body reserve, that is, stimulation lipid catabolism.Herein, the long-chain carboxylic acid's class discharged from adipose tissue
Catabolism occur in large quantities in entire body, particularly in liver, therefore, for improve endurance performance it is opposite
It is not the mitochondria only related in myocyte for this aspect of the invention.
The purposes of collagenic hydrolysate of the invention especially includes non-therapeutic use, that is, is administered to collagenic hydrolysate
Do not need medically endurance performance or weight in terms for the treatment of human or animal.On the contrary, the purpose of the purposes one side is
It typically would be desirable to endurance performance enhancing.This can contribute to improve quality of life, especially related with the people to be keen on sports.Separately
On the one hand, due to stimulation lipid catabolism caused by weight loss may mainly due to cosmetic reasons and cater to the need, that is,
Improve Body proportion.
However, in addition to this, the invention also includes collagenic hydrolysates for preventing and/or treating to be characterized in that mitochondria
The therapeutic use for the pathological conditions that activity reduces.In particular, the pathological conditions can with endurance show reduce and/
Or weight gain is characterized.
In the case where this therapeutic use, the pathological conditions are preferably selected from obesity, cardiovascular disease, the rhythm of the heart
Uneven, cardiac insufficiency, hypotony, hypertension, metabolic disorder, diabetes, metabolic syndrome, abrasive grit cellulous anemia, kidney
With the dysfunction of liver, neuropathy, incoordination, epileptic attack, dementia, Alzheimer's disease, autism, depression, slow
Fatigue syndrome, Parkinson's disease, motor neuron disease, multiple sclerosis, apoplexy sample symptom, migraine, myoclonia, fiber crops
Numbness, neuralgia, hyperalgia, hyperesthesia, dysphagia, vomiting, constipation, diarrhea, optic nerve fiber and retina denaturation,
Nystagmus, ptosis, yctalopia, hearing loss, deafness and inner ear disorders.For these indications, increase can be passed through
Mitochondria number improves mitochondria activity realization therapeutic effect.
Because the increase of mitochondria activity declines glucose level, the application of collagenic hydrolysate also to prevention and/or
Treating cancer, i.e. malignant tumour, there is good effect.This be based on following observation as a result, i.e. tumour cell mainly pass through it is scarce
Oxygen metabolism (lactic fermentation) supplies energy required for them, and is therefore necessarily dependent on glucose.Therefore, soma
Higher glucose consumption can weaken tumour cell.
In the case where the present invention, it was found that the protein kinase (AMPK) that collagenic hydrolysate causes enzyme AMP- to activate is more
Height expression.This adjusting enzyme equally has an impact to the energetic supersession of cell, so, increase AMPK quantity also to endurance performance and rouge
Matter catabolism has positive effect.The mitochondria activity caused by collagenic hydrolysate increases may between AMPK expression increase
There are direct correlations.
All use of the invention on the way, collagenic hydrolysate is preferably applied through enteron aisle, is especially administered orally.
In a preferred embodiment of the invention, collagenic hydrolysate is applied in the form of nutritional supplement.It is especially advantageous
, apply the form in the form of a solution for example as the bottle of preparation and carry out or carry out in powder form.Since it is good
Solubility, collagenic hydrolysate can also be added in different beverages without causing muddiness.Collagen water tasteless
The receiving that user can be improved in the application of solution product is horizontal.
Preferred embodiment according to the invention, other than collagenic hydrolysate, the nutritional supplement is without in addition
Protein or protein hydrolysate.It is formed in the nutritional supplement kept with muscle, is used for muscle known
Various protein, the people to be especially keen on sports, the purpose is to largely replace carbohydrate and rouge by protein
Fat is as energy supply object.However, purposes of the invention is not based on function of the collagenic hydrolysate as energy supply, but
Based on the above-mentioned specific function to mitochondria activity.
Therefore, in another embodiment of the present invention, other than collagenic hydrolysate, the nutritional supplement is not
Containing other physiologically active component.
However, optionally, the present invention also includes using collagenic hydrolysate as the (battalion with a variety of other components
Support) component of enriching substance is come the case where application.Specifically, collagenic hydrolysate can be added to food or snacks is for example skilful
Gram force stick, protein rod or cereal bars (so-called functional food);Or milk, dairy products (such as Yoghourt) and milk replacer (example
Such as soya-bean milk, almond milk and coconut milk) in.
No matter apply in what manner, collagenic hydrolysate is more excellent typically with daily 1-40g, preferably daily 2.5-30g
Select daily 10-25g, the amount application of especially daily 12.5-20g.
In collagenic hydrolysate not as unique physiologically active group timesharing of nutritional supplement, for use of the invention
For way, it can for general health and be especially presented with other components of positive effect with one or more to endurance
It is combined.Such component be preferably selected from vitamin C, vitamin B, D, E and K series, conjugated linoleic acid, caffeine and its
Derivative, Guarana extract, green-tea extract, Epigallo-catechin gallate (EGCG), creatine, L-BETAIN, L- melon ammonia
Acid, L-arginine, alpha-lipoic acid, N-acetylcystein, NADH, D-ribose, magnesium aminosuccinate, antioxidant such as anthocyanidin,
Carotenoid, flavonoids, resveratrol, glutathione, superoxide dismutase and xanthone such as mangiferin, minerals are such as
Iron, magnesium, calcium, zinc, selenium and phosphorus and other oroteins, hydrolysate or peptide such as soybean, wheat or lactalbumin.
Another advantageous embodiment of the invention is related to collagenic hydrolysate and 2,3-dimethoxy-5-methyl-6-decaprenyl-1,4-benzoquinone and/or panthenol ---
That is ubiquinone10Oxidation and reduction form combination, wherein due to panthenol better bioavilability and it is preferred that panthenol.Even if making
With the daily intaking amount of 50-100mg panthenol, the positive effect for physical efficiency is also observed, presumption is the antioxidation pair of panthenol
The promotion of mitochondria activity.In this way, collagenic hydrolysate is in the above-mentioned indication with mitochondria dysfunction
Effect is supported.Alternatively or additionally, it is also possible for combining collagenic hydrolysate with pyrroloquinoline quinone (PQQ),
PQQ is found to be a kind of important redox confactor recently.
In one embodiment of the invention, by the application of collagenic hydrolysate and endurance training or altitutde training
It is combined.The aerobic capacity of metabolism can be improved in endurance training.It is also known that in environment of the oxygen with respect to reduction
In physical training (hypoxic training) remarkable effect is presented with to endurance, so, while being applied using the dosage of collagenic hydrolysate
With, can correspondingly be expected synergistic effect.This is especially significant for the people to be keen on sports.
On the other hand, within the scope of the present invention, applied in the case where no endurance training, altitutde training or muscular training
Collagenic hydrolysate is equally possible and actually useful.In particular, showing in animal experiment (seeing below)
The influence having been observed that Ben Faming to mitochondria activity etc. is combined with normal body movement.
According to the present invention, the molecular weight of the collagenic hydrolysate used can change in a wide range, wherein the upper limit by
Collagenic hydrolysate has sufficiently high hydrolysis degree not at room temperature to be gelled and to provide for water solubility, this and denaturation
Collagen or gelatin are different.The soluble peptide of collagenic hydrolysate can be fully subsequently reabsorbed into body.Typically, collagen water
The average molecular weight that product has 200-25000Da is solved, preferably 1000-6000Da, more preferable 1200-4000Da are even more excellent
Select 1500-3500Da, especially 2800-3300Da.
Advantageously, producing collagenic hydrolysate by the way that enzymatic hydrolysis includes the starting material of collagen.For the hydrolysis,
Especially using microorganism or the endopeptidase and/or exopeptidase of plant origin.
The starting material comprising collagen is typically chosen from the skin or bone of vertebrate, preferably mammal, especially
The skin (ox-hide or pigskin) of ox or pig.Can be from these starting materials with One-step production collagenic hydrolysate, or it can be through
The intermediate stage of gelatin is spent, in this case, type A gelatin and type B gelatin can be used.
As alternative, the collagenic hydrolysate for purposes of the present invention can be produced by recombinant gene expression.
By using natural collagen sequence, collagen sequence especially from ox or pig and its gene modification cell (for example,
Yeast, bacterium or plant cell, especially tobacco cell) in expression, the product of production can be with corresponding comprising collagen
The hydrolysate of raw material is substantially the same.Herein, it is possible to obtain relatively narrow or precisely predetermined molecular weight distribution.Make
For alternative, the sequence can be changed by mutation, so that the special properties to product have an impact.
The invention further relates to for improving endurance performance and/or stimulation lipolytic generation by improving mitochondria activity
The method thanked, particularly lost weight.The method preferably includes through enteron aisle, particularly collagen water is administered orally to human or animal
Solve product.The method can be therapeutic method or actually non-therapeutic method.
The present invention is explained in greater detail with reference to the in vitro and in vivo experimental result described in the examples below.
In the accompanying drawings:
Fig. 1 shows the fluorescence microscope images for the SH-SY5Y cell cultivated in the presence of collagenic hydrolysate.
Embodiment
1. improving mitochondria number using collagenic hydrolysate
There is a possibility that showing collagenic hydrolysate in vitro with human nerve cell (Human Neuroblastoma Cell Line SH-SY5Y)
For improving the validity of mitochondria number.
By SH-SY5Y cell 0.05 weight %, 0.2 weight % and 2.5 weight % with various concentration collagen water
It solves and is cultivated in the culture medium of product.For this purpose, using the average molecular weight produced by enzymatic hydrolysis from pigskin gelatin in 3000Da
The collagenic hydrolysate (hereinafter referred to as collagenic hydrolysate A) of left and right.Pass through point for the peptide that gel permeation chromatography measures
Son amount distribution is shown in table 1 below:
Table 1: the molecular weight distribution of collagenic hydrolysate A
Fraction | Weight % |
>7500Da | <10 |
3500–7500Da | 20-30 |
1500–3500Da | 30-33 |
500–1500Da | 25-38 |
100–500Da | <5 |
In order to direct estimation mitochondria number, mitochondrial protein ingredient TOM20 is subjected to fluorescent marker.TOM20
It is the subunit of one of mitochondrial outer membrane receptor complex, has and cytoplasmic precursor protein (propetide) is moved to line grain
Function in body.There, the protein as the enzyme of respiratory chain or citrate cycle falls presequence by cracking and is activated.
As a result, under fluorescence microscope the quantity of the TOM20 of visible fluorescent marker be cell Mitochondria number amount
Degree.It is shown with the cell of the collagenic hydrolysate culture of 0.05 weight %, 0.2 weight % and 2.5 weight % in Figure 1A, 1B and 1C
In, wherein the clearly visible light fluorescence (being originally used for green) in nucleus (being originally used for blue) peripheral region is with concentration
Increase and increases.Thus, collagenic hydrolysate increases SH-SY5Y cell Mitochondria number and thus makes total mitochondria living
Property increase.
2. enzyme AMPK is by collagenic hydrolysate Activated in Vitro
The protein kinase (AMPK) of AMP- activation participates in the energy supply in both adipose tissue and muscle systems.Because
AMP is formed when consuming ATP, the indicant of energy deficiency can be seen as.Thus, the expression of AMPK is activated from deposit
Fat energy reserve and play a role in glycolytic cycle.
In order to determine influence that collagenic hydrolysate expresses AMPK, person monocytic cell is being included into 0.5mg/ml collagen water
It solves and is cultivated 24 hours in the culture medium of product.After removing culture medium, RNA is extracted from cellular layer, and draw using specificity
Object measures the amount of AMPK-RNA by PCR.
Compared with the control without collagenic hydrolysate, AMPK-RNA increases significantly (more than 600 times).Equally from this
It was found that thus demonstrating the stimulation of energetic supersession of the collagenic hydrolysate to cell.
3. enzyme AMPK is activated in collagenic hydrolysate body
The positive effect for confirming that collagenic hydrolysate expresses AMPK in animal experiment body can also be used.
For this purpose, being equivalent to the collagenic hydrolysate of the amount of 10g people's equivalent dose for the daily feeding of mouse, continue 3 months.?
After mouse is put to death, musculus quadriceps is fully cut off, is rapidly frozen and grinds.Soluble protein is extracted from musculature, and
The amount of AMPK is measured by immunoassay (ELISA).
Compared with the control group for not receiving collagenic hydrolysate, the amount of AMPK increases 1.5-2 times.
4. the external influence that collagenic hydrolysate generates NADH
From the low energy form NAD of nicotinamide adenine dinucleotide+Form rich form of energy NADH+H+It is equivalent to generate
The energy of ATP form.Therefore it is the active indirect measurement of myocyte's Mitochondria.
For this test, human muscle cell is cultivated 6 days in the culture medium comprising 0.5mg/ml collagenic hydrolysate.?
After removing culture medium, triglycerides is extracted, and is measured with the help of enzyme glycerokinase and glycerol-3-phosphate dehydrogenase sweet
With NADH+H in oily three esters and glycerol+The energy of form release.
Compared with the control without collagenic hydrolysate, about 2 times of the amount increase of NADH.
5. increasing mitochondria density in vivo in rats
In preclinical study, it can prove in Rat Skeletal muscle system caused by as applying different collagenic hydrolysates
Mitochondria density dramatically increases (that is, the increase of mitochondria number and/or increase of mitochondria size).
The research pairThe male rat of IGS system (Charles River Laboratories, Sulzfeld)
It carries out, it is 300-400g that the rat, which is 64 day age weight when studying and starting,.Test group and control group respectively include six animals.
When studying beginning (t=0), the quadriceps muscle of thigh biopsy of every rat is obtained by fine needle aspiration.In surrounding
Time in, the animal of test group then receives the day of the respective collagenic hydrolysate (seeing below) of per kilogram Current body mass 200mg
Dosage (daily dose for being equivalent to the 15g for the people of 75kg weight).Collagenic hydrolysate is dissolved in the concentration of 20mg/ml suitable
It is applied in the tap water of amount and by stomach tube.The animal of control group respectively receive equivalent do not include collagenic hydrolysate originally
Water.
Other than above-mentioned collagenic hydrolysate A, the collagen water of double-layer fur gelatin has been used in other test group
Solve product B, average molecular weight 2000Da;With the collagenic hydrolysate C of double-layer fur gelatin, average molecular weight 3500Da,
It is all to be produced by enzymatic hydrolysis under all situations.The molecular weight distribution of all three hydrolysates is shown in the following table 2:
Table 2: with the molecular weight distribution of the weight % collagenic hydrolysate indicated
Fraction | Hydrolysate A | Hydrolysate B | Hydrolysate C |
>7500Da | <10 | <3 | 4-14 |
3500–7500Da | 20-30 | 10-20 | 18-25 |
1500–3500Da | 30-33 | 25-32 | 30-38 |
500–1500Da | 25-38 | 40-50 | 28-36 |
100–500Da | <5 | <15 | <7 |
Average molecular weight | 3000Da | 2000Da | 3500Da |
After (t=4 weeks) terminates in the test period, all rats are put to death, and obtain stock four again by fine needle aspiration
Head Muscle biospy tissue.In surrounding, rat body weight averagely increases about 30%, does not have significance difference between test group and control group
It is different.
In order to determine mitochondria density, as described in the literature with before and after the tem study test period
(referring to A.Glauert and P.Lewis: prepared by the biological sample for transmission electron microscope for the biopsy of acquirement
(Biological Specimen Preparation for Transmission Electron Microscopy),
Princeton Legacy Library,2014).It in all situations, is 15.17 by the individual size of muscle biopsy tissue
x 15.17μm(230μm2) surface portion digitlization and semi-quantitative assessment.Herein, using 10 from each biopsy
Average mitochondrial surface product of a independent sample measurement as total surface area ratio.It can be from the clearly visible of the inner membrance with ridge
Characteristic structural positions mitochondria.
The following table 3 show the mitochondria density in the test group animal using collagenic hydrolysate A development (μm2Line grain
Body/230 μm2Total surface area):
Table 3: the development of test group A at any time
Rat number | Test group A, t=0 | Test group A, t=4w |
1 | 9.82 | 11.97 |
2 | 10.36 | 17.44 |
3 | 6.84 | 12.36 |
4 | 8.15 | 17.46 |
5 | 8.03 | 15.61 |
6 | 6.18 | 13.29 |
Average value | 8.23 | 14.69 |
Standard deviation | ±1.63 | ±2.48 |
The result shows that application collagenic hydrolysate A surrounding causes the obviously increase of mitochondria density, it is average to increase
78.5%, have statistical significance (p=0.001).
Mitochondria density comparison between test group and control group by applying collagenic hydrolysate A, B and C also turns out
Similar discovery.In all cases, the result after 4 weeks is shown in the following table 4:
Table 4: test group A, B and C are compared with control group
In test group, mitochondria density average specific control group high by 56.6% (A), 59.5% (B) and 90.4% (C), knot
Fruit equally has statistical significance (p=0.001).For all test groups, the Cohen's of the measurement as effect size
D value is both greater than 2.5, therefore all shows very specific effect in each case.
In short, this research provides tangible proof, it was demonstrated that application collagenic hydrolysate causes in myocyte
Mitochondria density dramatically increase and thus mitochondria activity is increase accordingly.This effect can with separate sources (pig and
Ox) and the collagenic hydrolysate of molecular weight distribution confirm.
Claims (18)
1. collagenic hydrolysate is used to improve the purposes of endurance performance by increasing mitochondria activity.
2. collagenic hydrolysate is used to stimulate lipid catabolism by increasing mitochondria activity, the use especially lost weight
On the way.
3. the purposes of claims 1 or 2, wherein the purposes is non-therapeutic use, especially beautifying use.
4. the purposes of claims 1 or 2, wherein the purposes is to be characterized in that mitochondria activity drops for preventing and/or treating
It is low, especially it is characterized in that the therapeutic use of the pathological conditions of endurance performance decline and/or weight gain.
5. the purposes of claim 4, wherein the pathological conditions are selected from obesity, cardiovascular disease, cardiac arrhythmia, heart function
Incomplete, hypotony, hypertension, metabolic disorder, diabetes, metabolic syndrome, abrasive grit cellulous anemia, kidney and liver function barrier
Hinder, neuropathy, incoordination, epileptic attack, dementia, Alzheimer's disease, autism, depression, chronic fatigue syndrome,
Parkinson's disease, motor neuron disease, multiple sclerosis, apoplexy sample symptom, migraine, myoclonia, paralysis, neuralgia, the pain sensation
Allergy, hyperesthesia, dysphagia, vomiting, constipation, diarrhea, optic nerve fiber and the denaturation of retina, nystagmus, eyelid
Sagging, yctalopia, hearing loss, deafness and inner ear disorders.
6. the purposes of any one of preceding claims, wherein collagenic hydrolysate is applied through enteron aisle, is especially administered orally.
7. the purposes of claim 6, wherein collagenic hydrolysate is in the form of nutritional supplement, particularly with solution or powder
Form application.
8. the purposes of claim 7, wherein the nutritional supplement does not include other albumen other than collagenic hydrolysate
Matter or protein hydrolysate.
9. the purposes of claim 7 or 8, wherein the nutritional supplement does not include other other than collagenic hydrolysate
Physiologically active component.
10. the purposes of any one of claim 6-9, wherein collagenic hydrolysate is with daily 1-40g, preferably daily 2.5-30g, more
It is preferred that the amount of daily 10-25g, especially daily 12.5-20g are applied.
11. the purposes of any one of claim 1-8 or claim 10, wherein by collagenic hydrolysate and one or more components
Combination, described group is selected from vitamin C, vitamin B, D, E and K series, conjugated linoleic acid, caffeine and its derivative, melon drawing
Receive extract, green-tea extract, Epigallo-catechin gallate (EGCG), creatine, L-BETAIN, L-citrulline, L-arginine,
Alpha-lipoic acid, N-acetylcystein, NADH, D-ribose, magnesium aminosuccinate, antioxidant such as anthocyanidin, carotenoid, class
Flavones, resveratrol, glutathione, superoxide dismutase and xanthone such as mangiferin, minerals for example iron, magnesium, calcium, zinc,
Selenium and phosphorus and other oroteins, hydrolysate or peptide such as soybean, wheat or lactalbumin.
12. the purposes of any one of claim 1-8, claim 10 or claim 11, wherein by collagenic hydrolysate with it is excellent
The 2,3-dimethoxy-5-methyl-6-decaprenyl-1,4-benzoquinone and/or panthenol that choosing is applied with the amount of daily 50-100mg combine, and/or combine with pyrroloquinoline quinone (PQQ).
13. the purposes of any one of preceding claims, the wherein application with endurance training or altitutde training phase of collagenic hydrolysate
Combination.
14. the purposes of any one of claim 1-12, wherein the application of collagenic hydrolysate is instructed on endurance training, plateau
It carries out in the case where experienced or muscular training, is especially combined with normal body movement.
15. the purposes of any one of preceding claims, wherein the average molecular weight of the collagenic hydrolysate is 200-
25000Da, preferably 1000-6000Da, more preferable 1200-4000Da, even more preferably 1500-3500Da, especially 2800-
3300Da。
16. the purposes of any one of preceding claims, wherein collagenic hydrolysate by the inclusion of the starting material of collagen enzymatic
Hydrolysis is to produce.
17. the purposes of claim 16, wherein described includes that the starting material of collagen is selected from vertebrate, preferably mammal
Skin or bone, and the skin of especially ox or pig.
18. the purposes of any one of claim 1-15, wherein collagenic hydrolysate is produced by recombinant gene expression.
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DE102017102873.0 | 2017-02-14 | ||
PCT/EP2017/071184 WO2018041684A1 (en) | 2016-08-30 | 2017-08-23 | Use of collagen hydrolysate for improving endurance performance and for stimulating lipocatabolism |
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