CN104666327A - Application of low-polymer mannuronic acid and derivative thereof in preparation of medicines and health products for resisting fatigue and amyotrophia - Google Patents
Application of low-polymer mannuronic acid and derivative thereof in preparation of medicines and health products for resisting fatigue and amyotrophia Download PDFInfo
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- CN104666327A CN104666327A CN201510018297.9A CN201510018297A CN104666327A CN 104666327 A CN104666327 A CN 104666327A CN 201510018297 A CN201510018297 A CN 201510018297A CN 104666327 A CN104666327 A CN 104666327A
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- resisting fatigue
- oligomannuronic
- oligomannuronic acid
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Landscapes
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Abstract
The invention provides an application of low-polymer mannuronic acid and a derivative thereof in preparation of medicines and health products for resisting fatigue and amyotrophia. With b-configuration marine acidic polysaccharide as a raw material, a water-soluble low-molecular weight mannuronic acid compound is obtained. An application of low-molecular weight mannuronic acid in preparation and treatment or improvement of fatigue and amyotrophia symptoms is proved through an experiment for the first time; the pharmacologic action of low-molecular weight mannuronic acid is very obvious; an experiment proves that low-molecular weight mannuronic acid is free of a toxic or side effect on muscle cells and is suitable for being taken for a long period of time. The low-molecular weight mannuronic acid provided by the invention is from a marine natural product, and has the advantages of being abundant in resource, easy to industrialize, high in safety, unique in structure and good in oral absorption, and has a wide market application prospect in resisting fatigue and amyotrophia.
Description
Technical field
The invention belongs to marine drug field, be specifically related to oligomannuronic acid and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof.
Background technology
Fatigue is the physiological acoustic signals process of body complexity, the normal physiological phenomenon of one that must occur when referring to mental or physical arrival certain phase.It had both indicated the temporary transient decline of the original ability to work of body, may be again the tendency that body develops into morbid state.Tired appearance, can cause motor capacity reduction, work efficiency reduces, mistake and accident increases, fighting capacity goes down.Can not get timely recovery after occurring if tired, accumulate gradually, also can cause " overwork ", make body generation endocrine regulation, immunity degradation, even occur organic disease, threaten human health.Organism fatigue by having a rest, sleeping and physical method elimination, can also realize the effect of resisting fatigue in addition by extra-nutrition material and pharmacotherapy.Some branched-chain amino acid (BCAA) comprise leucine and isoleucine etc., catabolism in muscle, are easily oxidized to body and provide energy, to maintaining the normal activity of human brain and delaying and eliminate sports fatigue to play an important role.In addition, creatine, vitamin B group, vitamin C, vitamin E, vitamin A, carotene, trace element-selenium, copper, zinc and metallothionein etc. are also had also can to improve the metabolism of body, the generation of delay fatigue.In recent years, the tired concern also more and more enjoying people as subhealth symptom, the generation of resisting fatigue and delay fatigue and/or the elimination of promotion fatigue, become the hot issue that sports medical science is studied at present.
Amyotrophy is that skeletal muscle volume compares with the state before age cohorts or self muscle the description occurring decline state.Under amyotrophy often occurs in old and feeble relevant damage and various malignant disease condition, if serious disease is as acquired immune deficiency syndrome (AIDS), cancer, congestive heart disease etc.Amyotrophy is many and perineural Motor nerve fibre damages and muscle fiber primary affection is relevant, from being divided into neurogenic and muscle-derived two type clinically, causes the amyotrophy of whole body diffusivity or local.The change of thin muscle quantities often can cause the balance of albumen synthesis and degraded in muscle fiber to change.Research display: short-term heavy dose or long-term super physiological dose use glucocorticoid to cause amyotrophy, its typical characteristic is that protein content declines (mainly fast muscle fiber), and muscle fiber reduces, and muscular strength declines, fatiguability.The health of amyotrophic generation meeting patient brings very large impact, in recent years along with the development of society, amyotrophic sickness rate is in the trend constantly risen, treatment amyotrophy also result in showing great attention to of people, current Therapeutic Method mainly contains Chinese and western medicine Drug therapy, dietary adjustments and motion and other physiotherapys, but these means reach far away desirable therapeutic effect at present, therefore, safe, efficient, economic anti-amyotrophy medicine is researched and developed and functional food has become current study hotspot.
Oligomannuronic acid is a kind of intercellular polysaccharide obtained by extraction and isolation in marine brown, is the ol cpds connected to form by Isosorbide-5-Nitrae-glycosidic bond by beta-D-mannuronic acid (β-D-Mannuronic acid, M).At present, bibliographical information oligomannuronic acid is had to have the various biological such as antioxidation, antiinflammatory, immunomodulating and anti-senile dementia both at home and abroad active, but at present, there is not yet report about the investigation and application of oligomannuronic acid oligosaccharide in resisting fatigue and anti-amyotrophy.
Summary of the invention
The object of this invention is to provide oligomannuronic acid and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, the present invention proves that oligomannuronic acid and derivant thereof have remarkable protective effect to exercise fatigue and muscle fiber atrophy on animal and cellular level by experiment.Compound of the present invention possesses the advantage of the acid oligosaccharide of low-molecular-weight, and not only water solublity absorbs by force, easily, and abundance, mechanism of action uniqueness, has wide industrialization development application prospect.
For achieving the above object, the present invention adopts following technical proposals to be achieved:
Oligomannuronic acid and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof.
The molecular skeleton of described oligomannuronic acid and derivant thereof is the linear oligosaccharides compound that D-MANNOSE aldehydic acid is connected to form by β-Isosorbide-5-Nitrae glycosidic bond, and weight average molecular weight is 0.3kDa ~ 12kDa; Wherein, the general formula of molecular structure of oligomannuronic acid and derivant thereof is as follows:
In formula, R=H or Li, Na, K, Ca, Mg or corresponding sulphation, phosphorylation, nitrification, carboxy methylation deriveding group, n≤60.
R '=H or corresponding sulphation, phosphorylation, nitrification, carboxy methylation deriveding group.
The consumption of described oligomannuronic acid and derivant thereof is 5mg/kg-100mg/kg.
Described oligomannuronic acid and derivant thereof significantly increase the tolerance of exercise of skeletal muscle.
Described oligomannuronic acid and derivant thereof can reduce the accumulation of serum urea nitrogen in the rear body of motion, reduce blood lactase acid accumulation in the rear body of motion.
Described oligomannuronic acid and derivant thereof can increase hepatic glycogen and muscle glycogen content.
The protein level of skeletal muscle content that described oligomannuronic acid and derivant thereof can suppress glucocorticoid to cause declines.
Low-molecular-weight mannuronic acid extracts the Algin obtained and obtains through classification and degraded from Thallus Laminariae (Thallus Eckloniae), Macrocystis pyrifera (L.) Ag., Fucus Vesiculosus, yellow tang, Thallus Laminariae (Thallus Eckloniae) or Chorda filum (L.) Stackh. are correlated with Brown algae.
Described low-molecular-weight mannuronic acid and derivant thereof and anti-fatigue medicament Radix Ginseng class, Radix Et Caulis Acanthopanacis Senticosi, Lac regis apis, adenosine triphosphate, Radix Rhodiolae, astaxanthin, spirulina, Cordyceps class, vitamins are composite, or with anti-amyotrophy medicine neurotrophic peptide class, or with comprise the Algin of oligomannuronic acid salt, the composite formation medicine of brown alga oligose or health product.
Advantage of the present invention and beneficial effect are:
1, the present invention adopts swimming power to exhaust animal model and skeletal muscle atrophy cell model, verify that oligomannuronic acid and derivant thereof are in resisting fatigue and anti-amyotrophic effect by experiment, research proves, oligomannuronic acid salt can significant prolongation swimming time, the deposit of effective increase hepatic glycogen and muscle glycogen, the remarkable level reducing the rear serum urea of motion and blood lactase acid, can make mice body endurance strengthen, bear sports load ability and strengthen; Significantly improve Skeletal Muscle Cell protein content simultaneously, significantly suppress the loss degraded of myotube protein, thus realize resisting fatigue and anti-amyotrophic effect.Oligomannuronic acid of the present invention and derivant thereof can be used for prevention or treatment is tired and amyotrophy symptom.
The plurality of advantages such as 2, oligomannuronic acid salt of the present invention derives from marine natural polysaccharide, has aboundresources, is easy to industrialization, safe and effective, have wide development prospect in resisting fatigue and anti-amyotrophy.
Detailed description of the invention
Below in conjunction with detailed description of the invention, technical scheme of the present invention is described in further detail.
The invention provides the purposes of low-molecular-weight mannuronic acid and derivant thereof, for the preparation of resisting fatigue and anti-amyotrophic medicine and health product.The present invention proves that low-molecular-weight mannuronic acid and derivant thereof can extend the swimming time of Exhausted mice effectively, significantly reduce the level of blood lactase acid and serum urea nitrogen, and significantly improve the hepatic glycogen of Exhausted mice and the content of muscle glycogen by experiment first; The simultaneously minimizing of the low-molecular-weight mannuronic acid myotube protein that can significantly suppress glucocorticoid to cause and α-actin content, and effectively suppress the degradation process of myotube protein; Therefore can be used for prevention, treatment or improve tired and muscle wasting symptoms.
Low-molecular-weight mannuronic acid in the present invention can extract the Algin obtained and obtain through classification and degraded from Thallus Laminariae (Thallus Eckloniae), Macrocystis pyrifera (L.) Ag., Fucus Vesiculosus, yellow tang, Thallus Laminariae (Thallus Eckloniae) or Chorda filum (L.) Stackh. are correlated with Brown algae, also can be synthesized into through chemical method.Described oligomannuronic acid salt is the lithium salts of the oligomannuronic acid of different molecular weight, sodium salt, potassium salt, calcium salt or magnesium salt, the molecular skeleton of described oligomannuronic acid and derivant thereof is that D-MANNOSE aldehydic acid (M) is by β-1, the linear oligosaccharides compound that 4 glycosidic bonds are connected to form, by β-1 between saccharide residue, 4-D-mannuronic acid forms, and structural formula is as follows:
Its weight average molecular weight range is 0.3kD ~ 15kD (being preferably 1kD ~ 10kD), n≤60 (being preferably 1 ~ 30), in formula, R=H or Li, Na, K, Ca, Mg or corresponding sulphation, phosphorylation, nitrification, carboxy methylation deriveding group, R '=H or corresponding sulphation, phosphorylation, nitrification, carboxy methylation deriveding group.
Exhaust animal pattern and glucocorticoid inducible amyotrophy model cell according to the tired power of classics, have rated oligomannuronic acid and derivant thereof to impact that is tired and muscle wasting symptoms.
Embodiment 1: oligomannuronic acid salt (LM) is to the protective effect of disposable swimming Exhausted mice
1) LM is on the impact of Exhausted mice swimming time
Mice swimming power is adopted to exhaust model namely: the continuous gavage of mice is after 2 weeks, positive drug selects octacosanol (10mg/kg), oligomannuronic acid salt is divided into low dose group (25mg/kg), middle dosage group (50mg/kg), high dose group (100mg/kg), after processing half an hour for the last time, the galvanized wire of 3% body weight that mouse tail is born a heavy burden, be placed in the swimming pool went swimming of 50 × 50 × 40 (unit is cm), water temperature 30 ± 1 DEG C, record mice starts to swim to swimming not power and exhausts the time of sinking to underwater can not emerge for 8 seconds, namely mice swimming power exhausts the time.
Result is as shown in table 1: measure mice persistent movement can reflect body endurance to the time that power exhausts.As shown in Table 1, there is dose-effect relationship in swimming time and LM, high dose group increases at most relative to blank group swimming time, 90.5%, median dose group increases by 66.2%, octacosanol group adds 67.5%, shows that LM high dose group and middle dosage group can improve the exercise tolerance of mice, has antifatigue effect.
Table 1 LM is on the impact of mice burden swimming time
* p<0.01, compares with model group.
2) LM is on the impact of Exhausted mice blood lactase acid (BLA) and serum urea nitrogen (BUN)
Mice swimming power is adopted to exhaust model namely: the continuous gavage of mice is after 2 weeks, positive drug selects octacosanol (10mg/kg), oligomannuronic acid salt is divided into low dose group (25mg/kg), middle dosage group (50mg/kg), high dose group (100mg/kg), after processing half an hour for the last time, tail vein gets blood, measure the front blood urea nitrogen (BUN) of swimming, blood lactase acid (BLA) content, the galvanized wire of 3% body weight that mouse tail is born a heavy burden, be placed in the swimming pool went swimming of 50 × 50 × 40, water temperature 30 ± 1 DEG C, swim after 1 hour and take out, remove and bear a heavy burden, after quiet 15min, eyeball gets blood, measure Mouse Blood blood urea nitrogen (BUN), blood lactase acid (BLA) content.Usually causing tired biochemical indicator by measuring, tired appearance and evolution can be observed.When mice body does not obtain abundant energy by sugar, catabolism of fat for a long time, protein and amino acids catabolism just can be strengthened, and the metabolism of nucleotide is also accelerated thereupon, and the end product of this metabolic pathway all generates carbamide.Plasma wrea produces fewer, illustrates that mice body endurance is stronger, more can bear sports load.
From table 2, before swimming, each group serum urea nitrogen difference is not obvious, and after swimming, each group serum urea nitrogen content obviously increases.After swimming, compared with model group, in LM, dosage and high dose group and octacosanol group serum urea nitrogen are significantly lower than model group, and this says that LM can reduce the accumulation of serum urea nitrogen in exercised rats body.
Blood lactase acid is glucolytic product under anoxic conditions, and along with the quickening of glycolysis speed, in muscle, the content of lactic acid is constantly accumulated, and causes Cellular pH value to decline, and produces a series of Biochemical changes, and then causes fatigue.Accumulate more lactic acid in muscle, tired performance degree is more serious.As can be seen here, before swimming, each group Serum lactic acid content difference is not obvious, and after swimming, each group Serum lactic acid content obviously increases.And model group Serum lactic acid content is apparently higher than high dose group in LM and octacosanol group, illustrates that LM can reduce blood lactase acid accumulation in body after mouse movement, accelerate the recovery of body.
Table 2 is on the impact of mice serum blood urea nitrogen (BUN) and blood lactase acid (BLA)
* p<0.05, compares with model group
3) LM is on the impact of Exhausted mice hepatic glycogen and Body development
Mice swimming power is adopted to exhaust model namely: the continuous gavage of mice is after 2 weeks, positive drug selects octacosanol (10mg/kg), oligomannuronic acid salt is divided into low dose group (25mg/kg), middle dosage group (50mg/kg), high dose group (100mg/kg), after processing half an hour for the last time, the galvanized wire of 3% body weight that mouse tail is born a heavy burden, be placed in the swimming pool went swimming of 50 × 50 × 40, water temperature 30 ± 1 DEG C, swim after 1 hour and take out, remove and bear a heavy burden, after eyeball gets blood, dislocate execution immediately, take out liver and hind leg muscle, measure hepatic glycogen (LG) and muscle glycogen (MG) content respectively.Because the deposit of glycogen in living organism and exercise tolerance are proportionate.When movement time increases, can consume muscle glycogen in a large number, living organism motor capacity is reduced, occur tired, add the intake for blood glucose simultaneously, when picked-up speed is greater than the decomposition rate of hepatic glycogen, blood sugar level reduces.Because blood glucose energy supply is to central nervous system, blood glucose reduction can cause central nervous system to be short of power, thus causes whole body fatigue.
Experimental result (table 3) shows, the hepatic glycogen of middle and high dosage group and octacosanol group and muscle glycogen are apparently higher than blank group, illustrate that the LM of middle and high dosage group can increase hepatic glycogen and muscle glycogen content, and glycogen can become glucose to enter blood by Direct Resolution, utilized for its hetero-organization by blood circulation.Therefore, the LM of middle and high dosage group can delay and resist tired generation.
Table 3LM is on the impact of Mouse Liver glycogen (LG) and muscle glycogen (MG)
* p<0.05, * * p<0.01 compares with model group; #p<0.05, compares with positive drug.
Embodiment 2: oligomannuronic acid salt (LM) protects myocyte to avoid the effect of myotube atrophy
1) LM is on the impact of C2C12 myotube protein content
This experiment adopts mice skeletal C2C12 myoblast differentiation to form C2C12 myotube in 8 days, stimulates 24 hours, set up classical amyotrophy cell model with dexamethasone DEX (100nM).α-actin is the one of actin subclass, and be the albumen of sarcostyle rich content, its content affects the contractility of skeletal muscle.Researcher commonly uses the change of the albumen of α-actin to reflect the change of Skeletal Muscle Contraction albumen.Amyotrophy generation phase is with the decline of α-actin protein content.
The impact of LM on protein level of skeletal muscle content is observed in experiment.BCA method is adopted to detect muscle total protein, western Blot method is adopted to detect the content of α-actin, result is as shown in table 4, after dexamethasone DEX (100nM) processes myotubes 24 hours, total protein content is consistent with the downward trend of α-actin content, all there is significant difference, show that model is successfully established.The LM of test dose can increase total protein content and α-actin content significantly, and in dose-dependent effect.This shows the amyotrophy phenomenon that the protein level of skeletal muscle content that LM can suppress glucocorticoid to cause effectively declines.
Table 4 LM is on the impact of Skeletal Muscle Cell total protein content and α-actin content
##p<0.01, compares with blank group; * p<0.05, * * p<0.01 compares with model group.
2) LM impact that C2C12 myotube protein is degraded
This experiment adopts mice skeletal C2C12 myoblast differentiation to form C2C12 myotube in 8 days, stimulates 24 hours, set up classical amyotrophy cell model with dexamethasone DEX (100nM).Proteins ubiquitin degradation pathway is the main cause that skeletal muscle is lost.
LM is observed on the impact of myotube protein ubiquitination Degradation Level by western Blot method in experiment.Shown in result table 5, after dexamethasone DEX (100nM) processes myotubes 24 hours, the ubiquitination level of albumen significantly rises, and shows that the protein degradation of model cell is accelerated.The LM of test dose all can the ubiquitination degraded of Profilin significantly, and in dose-dependent effect.This shows the loss of the protein level of skeletal muscle that LM can suppress glucocorticoid to cause effectively, and dosage is when 100ug/ml, and suppression ratio, up to 35.7%, has good anti-amyotrophy effect.
Table 5 LM is on the impact of Skeletal Muscle Cell proteins ubiquitin degradation pathway
##p<0.01, compares with blank group; * p<0.05, * * p<0.01 compares with model group.
Experimental result of the present invention shows, low-molecular-weight mannuronic acid salt better can increase the tolerance of exercise of skeletal muscle, accelerates the recovery of tired body, obviously suppresses myotube protein to be lost and decomposes and reach resisting fatigue and anti-amyotrophic effect.This low-molecular-weight mannuronic acid salt can be used for prevention and therapy that is tired and amyotrophy disease as a kind of natural health-care products or medicine.These goods derive from ocean Sargassum, have aboundresources, are easy to industrialization, and safe advantages of higher, in resisting fatigue and anti-amyotrophy, there is wide market and application prospect.Higher value application for ocean Sargassum is offered reference meaning and enlightenment by the present invention.
Above embodiment only in order to technical scheme of the present invention to be described, but not is limited; Although with reference to previous embodiment to invention has been detailed description, for the person of ordinary skill of the art, still can modify to the technical scheme described in previous embodiment, or equivalent replacement is carried out to wherein portion of techniques feature; And these amendments or replacement, do not make the essence of appropriate technical solution depart from the spirit and scope of the present invention's technical scheme required for protection.
Claims (10)
1. oligomannuronic acid and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof.
2. oligomannuronic acid according to claim 1 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, it is characterized in that: the molecular skeleton of described oligomannuronic acid and derivant thereof is that D-MANNOSE aldehydic acid is by β-1, the linear oligosaccharides compound that 4 glycosidic bonds are connected to form, weight average molecular weight is 0.3 kDa ~ 12 kDa; Wherein, the general formula of molecular structure of oligomannuronic acid and derivant thereof is as follows:
In formula, R=H or Li, Na, K, Ca, Mg or corresponding sulphation, phosphorylation, nitrification, carboxy methylation deriveding group, n≤60;
R '=H or corresponding sulphation, phosphorylation, nitrification, carboxy methylation deriveding group.
3. oligomannuronic acid according to claim 1 and 2 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, is characterized in that: the consumption of described oligomannuronic acid and derivant thereof is 5 mg/kg-100 mg/kg.
4. oligomannuronic acid according to claim 1 and 2 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, is characterized in that: described oligomannuronic acid and derivant thereof significantly increase the tolerance of exercise of skeletal muscle.
5. oligomannuronic acid according to claim 1 and 2 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, it is characterized in that: described oligomannuronic acid and derivant thereof can reduce the accumulation of serum urea nitrogen in the rear body of motion, reduce blood lactase acid accumulation in the rear body of motion.
6. oligomannuronic acid according to claim 1 and 2 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, is characterized in that: described oligomannuronic acid and derivant thereof can increase hepatic glycogen and muscle glycogen content.
7. oligomannuronic acid according to claim 1 and 2 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, is characterized in that: the protein level of skeletal muscle content that described oligomannuronic acid and derivant thereof can suppress glucocorticoid to cause declines.
8. oligomannuronic acid according to claim 1 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, is characterized in that: described n is 1 ~ 30.
9. oligomannuronic acid according to claim 1 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, is characterized in that: low-molecular-weight mannuronic acid extracts the Algin obtained and obtains through classification and degraded from Thallus Laminariae (Thallus Eckloniae), Macrocystis pyrifera (L.) Ag., Fucus Vesiculosus, yellow tang, Thallus Laminariae (Thallus Eckloniae) or Chorda filum (L.) Stackh..
10. oligomannuronic acid according to claim 1 and the application of derivant in preparation resisting fatigue and anti-amyotrophic medicine and health product thereof, it is characterized in that: described low-molecular-weight mannuronic acid and derivant thereof and anti-fatigue medicament Radix Ginseng class, Radix Et Caulis Acanthopanacis Senticosi, Lac regis apis, adenosine triphosphate, Radix Rhodiolae, astaxanthin, spirulina, Cordyceps class, vitamins are composite, or with anti-amyotrophy medicine neurotrophic peptide class, or with comprise the Algin of oligomannuronic acid salt, the composite formation medicine of algin oligosaccharide or health product.
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