Background technology
Final dedust (AFG) is in ginseng concocting process, and fresh ginseng contracts at baking stage maltose and arginine generation carbonyl aldehyde and reacts to reset through Amadori and generates.Research finds that AFG not only has promotion microcirculation
[1], vasodilation
[2]with treatment diabetes
[3]deng pharmacologically active, also can promote that skin collagen synthesizes and improves skin elasticity
[4].Recording AFG route of synthesis in document is
[5]: under anhydrous glacial acetic acid environment, arginine and maltose mass ratio are 1:2,80 DEG C of reaction 0.5h ~ 1.0h, synthetic ratio is about 27%, and thick synthetics has intense stimulus tart flavour, therefore, is necessary to find the synthetic method that a kind of synthetic ratio is high, have no irritating odor.
Along with the development of world's aging trend and the increase of job and life stress, disease aging and subhealth state aging also become clear day by day.Except gene fragment disappearance and sudden change
[6-7]outward, oxidativestress damage
[8,9,10]cause old and feeble topmost reason.This experiment discussion AFG causes the restraining effect of mouse subacute aging to D-semi-lactosi, according to the pharmacokinetics of AFG in rat body
[11]known, the main metabolic position of AFG in body is liver, also brain is acted on by hemato encephalic barrier, therefore according to mouse aging body changes of biochemical indexes, its repair ability to mouse aging vivo oxidation stress damage is judged by the change detecting resistance of oxidation in AFG administration group mouse liver and cerebral tissue.
content of the present invention
AFG synthetic method: arginine and maltose with without hydrothermally stable solvent for reaction media, solid organic acid is catalyzer, generates final dedust (AFG) under heating and mixing condition.
The present invention causes mouse subacute aging model modeling position to D-semi-lactosi and best modeling dosage is optimized, draw: when D-semi-lactosi causes the modeling of mouse subacute aging model, nape portion hypodermic injection (SC) is better than intraperitoneal injection (IP), and its best modeling dosage is respectively 0.5g/kg and 2.5g/kg.
Disclose AFG (50mg/kg) by experiment in vivo, hydroxyproline content in the cerebral index of mouse aging, cerebral tissue total antioxidant capacity and skin can be significantly improved, significantly reduce mda and content of nitric oxide in cerebral tissue, effectively treat aging;
Disclosing AFG (30mg/kg) by experiment in vivo prevents administration can slow down mice age symptom, to improve in mouse thymus index, skin in hydroxyproline content, serum hydrogen peroxide enzyme level in superoxide dismutase levels, cerebral tissue total antioxidant capacity and liver, can be anti-aging effectively in advance.
positively effect of the present invention is:
The synthetic method of AFG in the present invention, synthetic ratio has and significantly improves, can 79% be reached, reduce the production cost of AFG, and thick synthetics has no irritating odor, synthesizing acidic substance used is common food additive simultaneously, glycerol is also foods and cosmetics additive, to all nontoxic nonirritant of human body, add the safety in utilization of the thick synthetics of AFG, the raising of synthetic ratio accelerates the paces that AFG moves towards the industrialization.
The present invention inquires into out D-semi-lactosi through preliminary experiment and causes the best modeling position of mouse subacute aging and best modeling dosage thereof, improve the science studying anti-ageing medicine to disclose AFG and have and improve body total antioxidant capacity, the oxidativestress damage of remarkable reparation body, improve senile organism cerebral index and thymus index, improve skin elasticity, general performance anti-aging effects.
the synthesis of experimental example one AFG and detection
The synthesis of 1.AFG
1.1 accurate weighing 1.0g arginine, 2.0g maltose and 0.75g citric acid, be placed in the anhydrous glycerol of 20ml, be heated to 85 DEG C under mixing condition, cools after reaction 2.5h ~ 3.0h, detects AFG concentration in thick synthesis after dilution.
1.2 accurate weighing reaction substrates (1.0g arginine+2.0g maltose) 4 parts, are all put in 20ml glycerol, add 1.5g, 1.0g, 0.75g and 0.5g citric acid successively, be heated to 85 DEG C, cool after reaction 2.5h.AFG concentration is detected after thick synthetics dilution.
1.3 accurate weighing reaction substrates (1.0g arginine+2.0g maltose) add citric acid 0.5g, and mixing is heated to 85 DEG C of reaction 2.5h cooling, detect the change of AFG synthetic ratio after thick synthetics dilution.
1.4 setting blank groups: 0.75g citric acid and 20ml glycerol 85 DEG C react 2.5h.
1.5 other solid organic acids (oxysuccinic acid, phenylformic acid and oxalic acid etc.) catalysis AFG building-up reactions, method is with this patent 1.1.
The detection of the thick synthetics of 2.AFG
Use column front derivation high performance liquid chromatography
[5], marking product with blank group (glycerol 20ml+ citric acid 0.75g, 85 DEG C of reaction 2.5h) and AFG is blank and contrast, detects and calculate the production rate of AFG in thick synthetics.)
Chromatographic condition: Venusil-AA amino acid analysis dedicated columns (5 μm, 4. 6 mm × 250 mm).Mobile phase A: sodium acetate buffer solution-acetonitrile solution (pH=6. 5); Mobile phase B: acetonitrile solution V (acetonitrile): V (water)=4: 1.0 min, 0%B; 4 min, 3% B; 16 min, 10% B; 17 min, 20% B; 32min, 34% B; 34 min, 100% B; 38 min, 0% B gradient elution; Flow velocity 1. 0 mL/min; Determined wavelength 254 nm; Column temperature 40 DEG C; Sample size 20 μ L.
result
As shown in accompanying drawing 1,2, AFG mark product and AFG+ARG mix mark and relatively can find out, the retention time of AFG does not change in mixed mark, can determine that the retention time of AFG and ARG under this testing conditions is respectively 5.9min and 14.3min.As shown in accompanying drawing Fig. 3, blank group without going out peak phenomenon, illustrates that glycerine and citric acid (phenylformic acid, oxysuccinic acid and oxalic acid etc.) can not produce the material of interference AFG and ARG mensuration under 85 DEG C of heating conditions in AFG and ARG retention time.
As shown in Figure 4: containing AFG in this patent method synthetics, and thick synthetics is diluted rear AFG can be detected directly.Illustrate that arginine and maltose generation Maillard reaction generate final dedust (AFG) under citric acid, glycerine and heating condition.This synthetic method is feasible, efficient, and synthetic ratio can reach 72.45%.
As shown in accompanying drawing 5 ~ 8, and the thick synthetics of citric acid relatively can be found out, under equal experiment condition, oxysuccinic acid, phenylformic acid, oxalic acid and succinic acid the reaction of catalysis arginine maltose can generate final dedust (AFG) equally.Can judge thus, in thermally-stabilised solvent, heat the solid organic acid presenting acidity all can catalysis Maillard reaction.
example two D-semi-lactosi causes the comparison at mouse subacute aging modeling position and the screening of corresponding dosage
1 laboratory animal
SPF level ICR ♂ mouse 80, at 2 monthly ages, purchased from Bethune medical college of Jilin University Experimental Animal Center, credit number is SCXK-(Ji) 2013-0003.Rearing conditions: temperature 23 ± 2 DEG C, humidity 55 ± 5%, feeds utilized is coventional type.
experimental technique
2.1 grouping and administrations (as shown in table 1)
The each component group of table 1 and administrations
2.2 study of behaviour detect
The observed and recorded mouse mental status, food ration, amount of drinking water, body weight and defecation situation in administration process.
2.3 biochemical indicators detect
Administration the 8th is dissected weekend, and in dissection administration in first 2 hours, before last administration, water is can't help in 12h fasting.Win eyeball to get blood and put to death, by the centrifugal 10min of 4200r/min under blood 4 DEG C of conditions, draw serum for subsequent use, extract brain, liver,spleen,kidney, thymus gland, weigh and calculate organ index after the cleaning of ice physiological saline.Cerebral tissue, serum and partial liver are stored under-80 DEG C of conditions with indices to be detected.
2.4 statistical analysis
Carry out processing data with SPSS17.0, result represents with `x ± s, and use one-way analysis of variance, P<0.05 thinks statistical significance.
result
3.1 mice behaviors are observed
As can be seen from the mental status, physiological site change, fur color and luster and excretion situation, compare ip group mice age degree with blank group and become dose-effect relationship with D-Gal, sc group mouse D-Ga low dose group mouse morphological specificity is without considerable change, and sc high dosage is more subcutaneous combines group mouse and show more obvious aging state.Therefore reach a conclusion: abdominal cavity aging that is middle and high and subcutaneous high dose group mouse is obvious.
3.2 impacts on organ index.
Index and spleen index as shown in Figure 9: Ip-L+NaNO2 group relative to blank group otherness extremely significantly (
p< 0.01) Ip-H group blank relative to Sc-H group comparatively significantly (
p< 0.01), illustrate that Ip-L+NaNO2 group, Ip-H group and Sc-H group Three doses all can make mice spleen enlargement.Spleen is one of peripheral immune organ, and splenomegaly is generally regarded as the sign of some disease
[12], such as tumour, hepatitis, acute and chronic infection etc.
As shown in attached 10, ip-M and nape portion sc-H causes mouse thymus atrophy.Thymus gland is the important lymphoid organ of body, shows the phenomenon (atrophy) more obviously changing with age growth-degenerate than other organ
[13], namely weight index shared by thymus gland reduces, and therefore, atrophy of thymus gland is regarded as the symptom of body aging.
3.4 on TG, TC in serum and the impact (as table 2, shown in 3) on cerebral tissue MDA, GSH
Blood lipid level (`x ± s, n=8) in table 2. serum
Table 2 illustrates that D-Gal all can cause TC and TG in mice serum by ip low dosage and ip high dosage and significantly raise, and D-semi-lactosi sc low dosage and sc high dosage also can cause TG in mice serum and significantly raise.Investigation and experiment display: hyperlipemia is the important factor to patients with aged metabolic syndrome kidney function damage
[14], therefore above Three doses can be used as old and feeble modeling dosage.
MDA content and GSH level (`x ± s, n=8) in table 3. cerebral tissue
Table 3 shows: use dosage in ip method injection D-gla cause micro-reduced glutathion in Mice brain tissues significantly reduce (
p< 0.01); SC method D-gla high dosage injection can cause group Mice Body in reductive glutathione content significantly reduce (
p< 0.05), mda content significantly raises (
p< 0.01).Glutathione content is the important indicator weighing mice age, its brain aging degree of horizontal indirect reaction in its cerebral tissue
[15].Mda is lipid peroxidation product, also Chang Zuowei Aging index, and its content increases with aging degree and increases
[16].
Comprehensive each dosage group draws the impact of serum, cerebral tissue and hepatic tissue: D-gla injects ip-M(2.5g/kg) and sc-H(500mg/kg/d) 56 days, the mouse aging obtained, global index more blank group of otherness is remarkable, and close with usual aging 12 monthly age above mouse index
[17,18,19], therefore can best modeling dosage using these two kinds of dosage as ip and nape portion sc.
the Antisenility Experiment of experimental example tri-arginine disaccharide glycosides
1 laboratory animal and material
1.1 laboratory animal (with experimental example two)
1.2 experiment material
Final dedust (AFG, laboratory is made by oneself, and purity is 99.84%), D-semi-lactosi (Shanghai Kai Yang Bioisystech Co., Ltd, analytical pure), nimodipine (Bayer Schering Pharma, 06170438).
method
2.1. animal grouping and process
After adaptability raises one week, mouse is divided into 7 groups at random.Be respectively blank, model, nimodipine, AFG treatment is low, AFG treatment is high, AFG prevention is low, AFG prevention is high.Advise according to pharmacological methods: every 10g injected in mice volume is no more than 0.2mL, then the administration volume (mL) of this experiment mice is 0.01 times of Mouse Weight (g).Except blank group, remain each group all in 1 ~ 8 week neck dorsal sc injection D-semi-lactosi 500mg/kg cause mouse subacute aging model, positive drug and AFG administering mode are per os gavage.
Table 4. is group dosage and administration time respectively
2.2 Indexs measure
Mouse was dissected in the 8th week, and dissect administration in first 2 hours, before last administration, water is can't help in 12h fasting.After eyeball gets blood, cervical dislocation is put to death, by 4200rmin under blood 4 DEG C of conditions
-1centrifugal 10min, draws Virus monitory SOD level.Extract brain, liver, thymus gland, calculating organ index of weighing after the cleaning of ice physiological saline.To detect in T-AOC, NO and MDA in cerebral tissue, liver HYD in CAT and GSH and skin.
2.3 statistical analysis
Carry out processing data with SPSS17.0, result represents with `x ± s, adopts one-way analysis of variance between group, and P<0.05 is for there being statistical significance.
result
3.1thymus index and cerebral index
Table 5 is group thymus gland and cerebral index situation (`x ± s, n=8) respectively
Thymus gland is the important lymphoid organ of body, and atrophy of thymus gland is the key character of body aging
[13].Cerebral tissue is the nervous center of body, the Behavioral change of regulation and control body, and old and feeble relevant Alzheimer's disease and cerebral index are closely connected [20].AFG can significantly improve cerebral index and the thymus index of mouse aging, illustrates to have certain restraining effect to aging, and low dosage (30mg/kg) act on thymus gland successful (
p< 0.01), and high dosage (50mg/kg) to brain effect significantly (
p< 0.01).
HYD content in SOD level and skin in serum
HYD content (`x ± s, n=8) in SOD level and skin in table 6. serum
Group |
Dosage (mg.kg-1) |
SOD (u.ml-1) |
HYD (μg.g-1) |
Contrast |
0.9%NS |
128.64±9.07 |
5.55±0.59 |
Model |
0.9%NS |
75.56±10.83## |
3.99±0.57
## |
Nimodipine |
16 |
73.26±5.37 |
4.41±0.72 |
Prevention low dosage |
30 |
67.76±12.36 |
6.52±0.53** |
Prevention high dosage |
50 |
69.91±11.61 |
5.52±0.74** |
Treatment low dosage |
30 |
115.73±14.63** |
5.30±1.22** |
Treatment high dosage |
50 |
134.29±12.17** |
6.42±0.59** |
SOD, CAT, GSH-Px and T-AOC etc. constitute body active oxygen system of defense jointly, are the important component parts of body defending system.Total antioxidant capacity embodies the defence capability of body, and closely bound up with the generation of aging-related disease, and improving total antioxidant capacity can effectively treat or delay senility
[9,10].As shown in table 3, each dosage group of AFG all can effectively improve SOD level in mouse aging serum (
p< 0.01) and skin in HYD (
p< 0.01) content, effect is better than positive drug nimodipine.This illustrates that AFG can improve the resistance of oxidation of mouse aging, and can improve skin elasticity, delay skin aging, demonstrates the cosmetic result of AFG
[4].
T-AOC, MDA and NO change in 3.3 cerebral tissues
T-AOC, MDA and NO situation (`x ± s, n=8) in table 7. cerebral tissue
Group | Dosage (mg.kg-1) | MDA(nmol.mgprot-1) | T-AOC(unit.ml-1) | NO((μmol.gprot-1)) |
Contrast | 0.9%NS | 8.15±3.95 | 71.61±14.57 | 3.64±0.91 |
Model | 0.9%NS | 13.98±6.76## | 15.62±7.65## | 5.16±1.75## |
Nimodipine | 16 | 11.80±4.3 | 37.81±11.00
** | 2.85±0.23** |
Prevention low dosage | 30 | 8.22±2.5** | 31.45±14.128
* | 3.68±0.75* |
Prevention high dosage | 50 | 8.13±2.19** | 49.88±14.50** | 3.43±0.76** |
Treatment low dosage | 30 | 8.11±2.59** | 65.52±4.32** | 3.04±0.78** |
Treatment high dosage | 50 | 5.51±1.84** | 67.47±12.38** | 3.65±1.15* |
As shown in table 4, AFG significantly reduce lipid peroxidation product (MDA) and content of nitric oxide in mouse aging cerebral tissue (
p< 0.01), significantly improve total antioxidant capacity in brain (
p< 0.01), and treatment group high dosage (50mg/kg) effect is better than low dosage (30mg/kg); Prevention group low dosage (30mg/kg) effect is better than high dosage (50mg/kg).In a word, AFG can improve cerebral tissue total antioxidant capacity, reduces the old and feeble brain injury brought to body, reduces the morbidity risk rate of alzheimer.
CAT level and GSH content in 3.4 livers
Table 8. AFG is on the impact of CAT and GSH in mouse aging liver
Group | Dosage (mg.kg-1) | CAT(U.gHb) | GSH(μmol.gprot-1) |
Contrast | 0.9%NS | 135.18±21.03 | 35.47±7.19 |
Model | 0.9%NS | 83.06±8.57## | 19.26±3.24
## |
Nimodipine | 16 | 116.11±24.03 | 30.90±8.11* |
Prevention low dosage | 30 | 123.90±12.49* | 19.51±1.96 |
Prevention high dosage | 50 | 115.95±33.09 | 23.00±4.33 |
Treatment low dosage | 30 | 124.16±15.69* | 22.82±4.33 |
Treatment high dosage | 50 | 116.98±19.05 | 31.30±7.84** |
Glutathione content is the important indicator weighing mice age, its liver aging degree of level height indirect reaction
[15].Mda is lipid peroxidation product, also Chang Zuowei Aging index, and its content increases with aging degree and increases
[16].As shown in table 5, AFG low dosage (30mg/kg) can significantly improve hydrogen peroxide enzyme level in liver (
p< 0.05), high dosage (50mg/kg) can significantly improve glutathione content (
p< 0.01), the main metabolic position of AFG is liver, and this illustrates that AFG can improve liver total antioxidant capacity, has liver protection effect.
Comprehensive each index draws: AFG can improve mouse aging body total antioxidant capacity and body immunity, reduces oxidativestress damage, protects liver and improve the functions such as skin elasticity.Low dosage AFG(30mg/kg) remarkable to the preventive effect of aging, and high dosage AFG(50mg/kg) stronger to the result for the treatment of of aging; AFG is low, high dosage all can improve mouse skin elasticity and reach the effect of delay skin aging, and overall testing index proves that AFG has anti-aging effects.
Specific embodiments
Embodiment 1
Accurately take arginine: maltose=1g:2g, add glycerol 10ml and citric acid 0.1, shaking table 160rpm mixes, and reacts 20min under 20 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 1.17%.
Embodiment 2
Accurately take arginine: maltose=1g:2g, add glycerol 10ml and citric acid 0.1, shaking table 160rpm mixes, and reacts 20min under 40 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 3.52%.
Embodiment 3
Accurately take arginine: maltose=1g:2g, add glycerol 10ml and citric acid 0.1, shaking table 160rpm mixes, and reacts 20min under 60 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 8.28%.
Embodiment 4
Accurately take arginine: maltose=1g:2g, add glycerol 10ml and citric acid 0.1, shaking table 160rpm mixes, and reacts 20min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 20.36%.
Embodiment 5
Accurately take arginine: maltose=1g:2g, add glycerol 10ml and citric acid 0.1, shaking table 160rpm mixes, and reacts 20min under 100 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 25.36%.
Embodiment 6
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.2, shaking table 160rpm mixes, and reacts 30min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 23.57%.
Embodiment 7
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.2, shaking table 160rpm mixes, and reacts 60min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 40.19%.
Embodiment 8
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.2, shaking table 160rpm mixes, and reacts 90min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 53.48%.
Embodiment 9
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.2, shaking table 160rpm mixes, and reacts 120min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 62.17%.
Embodiment 10
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.2, shaking table 160rpm mixes, and reacts 150min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 70.56%.
Embodiment 11
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.2, shaking table 160rpm mixes, and reacts 180min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 70.00%.
Embodiment 12
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.2, shaking table 160rpm mixes, and reacts 210min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 61.88%.
Embodiment 13
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.5, shaking table 160rpm mixes, and reacts 120min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 51.26%.
Embodiment 14
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.5, shaking table 160rpm mixes, and reacts 180min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 72.54%.
Embodiment 15
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 0.5, shaking table 160rpm mixes, and reacts 210min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 70.83%.
Embodiment 16
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 1.0g, shaking table 160rpm mixes, and reacts 180min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 56.28%.
Embodiment 17
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and citric acid 1.5g, shaking table 160rpm mixes, and reacts 180min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 58.17%.
Embodiment 18
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and oxysuccinic acid 1.0g, shaking table 160rpm mixes, and reacts 120min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 68.69%.
Embodiment 19
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and phenylformic acid 1.0g, shaking table 160rpm mixes, and reacts 120min under 80 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 70.26%.
Embodiment 19
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and oxalic acid 1.0g, shaking table 160rpm mixes, and reacts 120min under 85 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 60.75%.
Embodiment 20
Accurately take arginine: maltose=1g:2g, add glycerol 20ml and succinic acid 1.0g, shaking table 160rpm mixes, and reacts 90min under 85 DEG C of conditions, and column front derivation high performance liquid phase detects AFG content, and synthetic ratio is 42.19%.
Embodiment 21
Anti-ageing pulvis
AFG powder in Example 1 after purifying, vacuum, low pressure seal is bottled, and makes lyophilisate.Every bottle of AFG content is made to be 0.35g.Other requirements meet the Pharmacopoeia of the People's Republic of China 2010 editions relevant regulations about pulvis agent.
Embodiment 22
Anti-senility oral liquid
AFG powder after Example 1 purifying, is mixed with the aqueous solution of 35mg/ml, loads in vial after sterilizing, makes the oral liquid often propping up 10ml.Other requirements meet the Pharmacopoeia of the People's Republic of China 2010 editions relevant regulations about oral liquid.
Embodiment 23
Beauty and skin care water
The AFG powder of purifying in Example 1, adds in surfactant matrix liquid, and it is 200mg/100ml that AFG finally adds concentration.Other requirements meet China cosmetic production requirement specification.
reference
[1] Zheng Yinan, Zhang Jing, Okuda Takudox etc. arginine derivative is to microcirculatory effect [J]. Jilin Auto Industry, 1998,20 (4): 45-47.
[2]Ishihara Takafumi, Okuda Takudo. Preparation of arginylfructosylglucoses as vasodilators: Japan, 09316091[P].1997-12-09.
[3] Zheng Yinan. a kind of preparation method of arginine glucoside and the application in treatment diabetes thereof: China, 102178687 [P].
2011-09-14.
[4]Han Li-Kun, Saito Masato,Nakagawa Yukiko,et al. Antioxidant comprising arginyl-fructosyl-glucose,and use thereof: Japan, 2012140360[P].2012-07-26.
[5] Liu Limin. the research [D] of L-arginine and derivative thereof in Radix Panacis Quinquefolii. Jilin: Jilin Agriculture University, 2012.
[6]Kotoku Kurachi, Sumiko Kurachi, Kezhong Zhang,
etal.Molecular mechanisms of age-related regulation of genes[J].International Congress Series, 2004,1262:562-565.
[7]Jeannette Loram, Andrea Bodnar.Age-related changes in gene expression in tissues of the sea urchin Strongylocentrotus purpuratus[J].Mechanisms of Ageing and Development,2012,133:338-
347.
[8]Qingwei Ruan, Fang Liu, Zhanjuan Gao, etal. The anti-inflamm-aging and hepatoprotective effects of huperzine A in D-galactose-treated rats[J]. Mechanisms of Ageing and Development, 2013,134:89-97.
[9]Akiko Satoh, Takako Yokozawa, Eun Ju Cho, etal. Antioxidative effects related to the potential anti-aging properties of the Chinese prescription Kangen-karyu and Carthami Flos in senescence-accelerated mice[J].Arch. Gerontol. Geriatr,2004,39:69-82.
[10]Weiqi Zhong, Ning Liu, Yanguang Xie, etal.Antioxidant and anti-aging activities of mycelial polysaccharides from Lepista sordida[J]. International Journal of Biological Macromolecules, 2013,60:355-359.
[11] Wang Kun, Xu Xiaohua, Li Jiaqi, etc. final dedust (AFG) pharmacokinetic in rat body [J]. Jilin Auto Industry, 2014,36 (2): 2-21813.
[12] Dong Mei. the hepatosplenomegaly [J] that tumour is relevant with hemopathy. Chinese practical paediatrics magazine, 2004,19 (6): 333-334.
[13] Gong Zhangbin. thymus gland aging and immunosenescence [M]. foreign medical science. geriatrics fascicle, 2009,30 (4): 145-149.
[14] Liu Jie, Song Yuehong. hyperlipemia is on the impact [M] of old metabolic syndrome patients's kidney function damage. internal medicine, 2009,4 (4): 525-527.
[15] Li Xiaoping, first, Yao Jianhua, etc. the impact [J] on mouse aging gsh and total antioxidant capacity of. Margarita calcium in analogy training. Chinese pharmacology and clinical, 2001,17 (1): 21-22.
[16] Chen Yonglan, agriculture literary composition field, face Xiao Fang, etc. the impact [M] on mouse aging superoxide-dismutase and mda and hydroxy radical qiao of. Ligusticum wallichii. China and foreign countries' medical research, 2012,10 (4): 7-8.
[17] Zhu Yazhen, Zhu Hong light .D-semi-lactosi causes foundation and the detection method [N] thereof of aged animal model. Fudan Journal, 2007,34 (4): 617-619.
[18]Ji-Hun Park, Tae-Saeng. Choi. Polycystic ovary syndrome (PCOS)-like phenotypes in the D-galactose induced aging mouse model[J].2012,427:701-704.
[19] Chen Jun, Yang Guang, Liu Wenqun. mouse Antisenility Experiment and microorganism research on anti-senescence overview [J]. China brewages, 2011,6:13-16.
[20]Andras Bilkei-Gorzo. Genetic mouse models of brain ageing and Alzheimer's disease[J]. Pharmacology & Therapeutics,2013.12.009。