CN101455676A - Use of mussel polysaccharide in preparing anti-aging medicine or food - Google Patents
Use of mussel polysaccharide in preparing anti-aging medicine or food Download PDFInfo
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- CN101455676A CN101455676A CNA2009100448816A CN200910044881A CN101455676A CN 101455676 A CN101455676 A CN 101455676A CN A2009100448816 A CNA2009100448816 A CN A2009100448816A CN 200910044881 A CN200910044881 A CN 200910044881A CN 101455676 A CN101455676 A CN 101455676A
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Abstract
The invention relates to the technical field of medicament and discloses a new application of mussel polysaccharide separated from extract of marine life mussels in preparing anti-ageing medicament or foods. The mussel polysaccharide is extracted from fresh mussel meat of mytilus crassitestas in the East China Sea, can respectively improve activity of SOD and GSH-px in blood serum, livers and brain tissues and lower content of MDA. Therefore, the mussel polysaccharide has anti-ageing activity and can be used to prepare anti-ageing medicament or foods. The invention provides a new application to mussel polysaccharide which is helpful to explore and use marine life.
Description
Technical field
The present invention relates to medical technical field, is a kind of new purposes that the Mytilus Coruscus Polysaccharides that is separated to from the extract of marine organisms mussel is used to prepare antiaging agent or food.
Background technology
Mussel is the marine products bivalve shellfish, is commonly called as blue or green mouthful, Fructus mail micromali, is one of major product of China's sea-farming.Research [the Searching for Alternatives to Heparin Sulfated Fucans from MarineInvertebrate Paulo A.S.Moirao and Mariana S.Pereira 2000 that aspects such as separation and purification mannase, agglutinin and antibiotic substance from mussel are abroad arranged, Elsevier ScienceInc.All right reserved.1050-1738], studies show that mussel extract has antitumor, antiviral isoreactivity.The main active of mussel extract is a Mytilus Coruscus Polysaccharides, it has effect [the Hongli Xu of antitumor, antiviral, anti-liver injury and increase body immunity, Tingting Guo, Yi-fengGuo, Jianpeng Zhang, Yang Li, Weihua Feng, et al.Characterization andProtection on Acute Liver Injury of a Polysaccharide MP-I from MytilusCoruscus[J] .Glycobiology, 2008,18:97-103; Xu Hongli, Guo Tingting, Guo Yifeng, Zhang Jianpeng, Feng Weihua, JiaoBing Hua. the separation and purification of Mytilus Coruscus Polysaccharides MP-I and anti tumor activity in vitro research
J ". The 2nd Army Medical College journal, 2006,27 (9): 998-1001].But do not see as yet that so far relevant Mytilus Coruscus Polysaccharides has the report of activity of fighting against senium.
Summary of the invention
The invention provides the new purposes that a kind of Mytilus Coruscus Polysaccharides is used to prepare antiaging agent or food.
The preparation method of Mytilus Coruscus Polysaccharides of the present invention is as follows:
1. preparation mussel extract:
Get the fresh mussel meat of East Sea Mytilus crassitesta Lischke, with high speed agitator it is stirred homogenate, add routinely and use little fire instead behind the water boil and boil about 4-5h, filter to get filtrate, this is the mussel extracting solution;
2. preparation Mytilus Coruscus Polysaccharides:
After the mussel extracting solution that makes concentrated, the centrifugal precipitation of abandoning, the supernatant volume ratio is the sevage method gradation extraction of chloroform: n-butyl alcohol=4:1, lose protein precipitation until water, add the long-pending 90-95% ethanol of triploid again, stir, the room temperature standing over night, the centrifugal supernatant of abandoning will precipitate lyophilization, get Mytilus Coruscus Polysaccharides.
The sugared content of Mytilus Coruscus Polysaccharides and the mensuration of protein content:
The above-mentioned Mytilus Coruscus Polysaccharides 1g that makes is redissolved in the 100ml distilled water, and the centrifugal precipitation of abandoning is measured content sugared in the supernatant with sulfuric acid-phynol method respectively routinely, measure proteic content in the supernatant with the BCA method, testing result, the sugared content of Mytilus Coruscus Polysaccharides is 91%, protein content is 5%.
The zoopery of Mytilus Coruscus Polysaccharides activity of fighting against senium:
The life-span of SD rat calculated with the monthly age, it has been generally acknowledged that just to enter the old and feeble stage after 24 monthly ages, and therefore, this experiment is made as senile rat with 24 monthly age SD rats, and 3 monthly age SD rats are young rat.
Get 24 monthly age SD rats, be divided into old normal control group, Mytilus Coruscus Polysaccharides low dose group and Mytilus Coruscus Polysaccharides high dose group, and to establish one group of 3 monthly age SD rat be young control.Average food-intake according to 24 monthly age SD rat every days, press the different dosage of Mytilus Coruscus Polysaccharides, Mytilus Coruscus Polysaccharides and rodent feedstuff are mixed by a certain percentage, make mixed fodder respectively, make Mytilus Coruscus Polysaccharides high dose group rat by body weight oral 400mg/kg Mytilus Coruscus Polysaccharides every day, promptly the dosage of Mytilus Coruscus Polysaccharides is 400mg/kg/d; Mytilus Coruscus Polysaccharides low dose group rat is by body weight oral 200mg/kg Mytilus Coruscus Polysaccharides every day, and promptly the dosage of Mytilus Coruscus Polysaccharides is 200mg/kg/d, and aged control and young control rat be feeding rodent feedstuff then.Raised 90 days continuously, observe the organ index of rat heart, liver, kidney, spleen respectively; Its heart, liver, brain are carried out the sections observation pathology to be changed; Measure superoxide dismutase (SOD), malonaldehyde (MDA), glutathion peroxidase (GSH-px) level in rat blood serum, liver and the brain, measure the main biochemical indicator of serum, measure distribution and the amount of beta galactosidase at cerebral tissue.Experiment is found, compares with aged control, and the heart of Mytilus Coruscus Polysaccharides high dose group, liver, cerebral tissue pathological section HE dyeing are observed and do not seen that obvious aging sexually revises; The SOD level increases in its serum, liver, the cerebral tissue, and MDA content reduces; The GSH-px level increases in serum, the liver organization; The main biochemical indicator measurement result of serum shows in blood glucose (Glu), glutamate pyruvate transaminase (ALT), glutamic oxaloacetic transaminase, GOT (AST), blood urea nitrogen (BUN), inosine (CRE), cholesterol (CH) biochemical indicator that the Mytilus Coruscus Polysaccharides group is compared no difference of science of statistics with blue or green, aged control; The triglyceride of Mytilus Coruscus Polysaccharides group, young control, aged control (TG) level has significant difference.SABC is measured beta galactosidase and is found that in corticocerebral distribution and amount the aged control beta galactosidase is expressed more, and the expression in the Mytilus Coruscus Polysaccharides, young control rat cerebral cortex all obviously reduces.Experimental result shows that mussel Mytilus Coruscus Polysaccharides of the present invention has activity of fighting against senium, therefore, can be used for preparing antiaging agent or food.
Mytilus Coruscus Polysaccharides preparation method of the present invention is easy, has opened up a kind of new purposes for Mytilus Coruscus Polysaccharides, helps halobiontic development and use.
Description of drawings
Fig. 1. Mytilus Coruscus Polysaccharides is low, high dose group and green grass or young crops, the aged control SD rat heart, liver, brain tissue slice contrast collection of illustrative plates:
Fig. 1-A SD rat heart HE histological observation * 200 of dyeing
Fig. 1-B SD rat liver HE histological observation * 200 of dyeing
Fig. 1-C SD rat cerebral cortex HE histological observation * 200 of dyeing
Fig. 2. Mytilus Coruscus Polysaccharides is low, high dose group and green grass or young crops, aged control SD rat cerebral cortex beta galactosidase expression and the relatively collection of illustrative plates that distributes
Fig. 3. Mytilus Coruscus Polysaccharides is low, high dose group and green grass or young crops, the SOD of aged control SD rat cerebral tissue total activity comparison diagram
Fig. 4 Mytilus Coruscus Polysaccharides is low, the content comparison diagram of high dose group and green grass or young crops, the MDA of aged control SD rat cerebral tissue.
The specific embodiment
Now in conjunction with the accompanying drawings and embodiments, the present invention is described in detail.
Embodiment 1: the preparation Mytilus Coruscus Polysaccharides
1. material
East Sea Mytilus crassitesta Lischke is provided by mussel culture base, Sheng Shan town, Shengsi, Zhejiang Province.
2. reagent
Phenol, sulphuric acid, chloroform, ethanol (analytical pure), Shanghai chemical reagents corporation of Chinese Medicine group product.The glucose standard substance, SIGMA company product.BCA-100 protein quantification test kit can betting office be provided by the Shen, Shanghai.
3. instrument
BECKMAN CUOLTER J-251 type high speed centrifuge, BECKMAN CUOLTER company product.
The preparation Mytilus Coruscus Polysaccharides
1. prepare the mussel extracting solution
Get the fresh mussel meat 1000g of East Sea Mytilus crassitesta Lischke, with high speed agitator it is stirred homogenate, add routinely and use little fire instead behind the water boil and boil about 4h, filtered through gauze gets extracting solution;
2. preparation Mytilus Coruscus Polysaccharides
After the mussel extracting solution concentrated, low-speed centrifugal is abandoned precipitation, and the supernatant volume ratio is that the sevage method of chloroform: n-butyl alcohol=4:1 extracts 3 times, and water does not have protein precipitation and occurs, add 95% long-pending ethanol of triploid again, stir room temperature standing over night, the centrifugal supernatant of abandoning, to precipitate lyophilization and get Mytilus Coruscus Polysaccharides 51g, be the pale yellow powder shape, water absorption is strong, and dissolubility is good.
The sugared content and the determining the protein quantity of mussel extract and Mytilus Coruscus Polysaccharides
1. use the sugared content of spectrophotometry Mytilus Coruscus Polysaccharides:
(1) drafting of glucose standard curve: be mixed with the aqueous solution that concentration is respectively 50 μ g/ml, 100 μ g/ml, 200 μ g/ml, 400 μ g/ml, 800 μ g/ml with standard glucose earlier, use the phenol sulfuric acid process to carry out chromogenic reaction respectively then, reuse spectrophotometric determination OD value, with sugared concentration is abscissa, the OD value is a vertical coordinate, the drawing standard curve.
(2) measure the sugared content of Mytilus Coruscus Polysaccharides: Mytilus Coruscus Polysaccharides is mixed with the solution of 10mg/ml concentration, carries out chromogenic reaction with the phenol sulfuric acid process routinely, with spectrophotometric determination OD value, contrast the glucose standard curve then, must corresponding sugared concentration.The sugared content that calculates Mytilus Coruscus Polysaccharides is 91%.
2. measure the protein content of Mytilus Coruscus Polysaccharides with the BCA method:
Mytilus Coruscus Polysaccharides is mixed with the solution of 10mg/ml concentration, presses the operation of BCA method protein quantification test kit description, measure the protein concentration of Mytilus Coruscus Polysaccharides solution, the protein content that calculates Mytilus Coruscus Polysaccharides is 5%.
The activity of fighting against senium animal experiment of Mytilus Coruscus Polysaccharides
1. material
Mytilus Coruscus Polysaccharides is made by embodiment 1.
20 of 24 monthly ages of male SD rat, 5 of 3 monthly ages, provide by west, Shanghai pul-Bi Kai laboratory animal company [SYXK (Shanghai) 2003-0018], the Rodents forage feed, free diet, sub-cage rearing is in the Experimental Animal Center SPF of The 2nd Army Medical College level Animal House.
2. reagent
Formalin provides lot number GB685-93 by Shanghai Pacific Ocean chemical industry (group) company product.BCA-100 protein quantification test kit can betting office be provided by the Shen, Shanghai.Superoxide dismutase (SOD) test kit, malonaldehyde (MDA) test kit, glutathione peroxidase (GSH-px) test kit builds up bio-engineering research by Nanjing to be provided, and the beta galactosidase staining kit is provided by the outstanding U.S. gene in Shanghai Pharmaceutical Technology Co., Ltd.
3. instrument
Microplate reader Versa Max (Molecular Devices)
H-S electric-heated thermostatic water bath (Dongtai electrical apparatus factory)
Bechman coulter J-251 type high speed centrifuge (Bechman)
4. experimental technique and result
4.1 experimental technique
15 of 24 monthly ages of male SD rat are divided into 3 groups at random, i.e. aged control, Mytilus Coruscus Polysaccharides low dose group and Mytilus Coruscus Polysaccharides high dose group, 5 every group; 5 of 3 monthly age SD rats are as young control.
Aged control and young control rat feeding rodent feedstuff, Mytilus Coruscus Polysaccharides is low, high dose group is distinguished the feeding Mytilus Coruscus Polysaccharides and the rodent feedstuff is pressed the blended mixed fodder of corresponding proportion, the dosage of Mytilus Coruscus Polysaccharides low dose group is 200mg/kg/d, the dosage of Mytilus Coruscus Polysaccharides high dose group is 400mg/kg/d, raises continuously 90 days.
Observe following index: 1) ordinary circumstances such as activities in rats, reaction, hair color; 2) organ index of rat heart, liver, kidney, spleen; 3) the tissue slice pathology of heart, liver, cerebral tissue change; 4) SOD, MDA, GSH-px level in serum, liver and the cerebral tissue; 5) the main biochemical indicator of serum; 6) beta galactosidase is in corticocerebral distribution and amount.
4.2 result
4.2.1 animal ordinary circumstance:
The aged control rat is blunt few moving, and sluggish lacks susceptiveness, and hair is withered and yellow; Low, the high dose group of Mytilus Coruscus Polysaccharides, rat is dysphoria in the early stage, and the fluffy and disorderly few gloss of hair is vivaciously active gradually subsequently, is quick on the draw, and hair is still smooth; The young control rat is obviously vivaciously active, is quick on the draw, and hair is bright and clean to be white in color.
4.2.2 organ index measurement result:
So-called organ index is meant ratio (%)=internal organs weight (the g)/body weight (g) * 100% of internal organs and body weight.Analyze by statistics, each organizes the heart, liver, the kidney of rat, the organ index of spleen sees Table 1.
Table 1 SD Rats Organs and Tissues index result (x ± s, n=5, unit: %)
*P＜0.05 is with aged control
By table 1 as seen, the liver index that removes young control than aged control height (significant difference is arranged,
*P＜0.05) outside, surplus each organ index does not have group difference.Mytilus Coruscus Polysaccharides does not cause the hypertrophy or the atrophy of organ, points out Mytilus Coruscus Polysaccharides not have internal organs toxicity indirectly.
4.2.3 the SD rat heart, liver, brain tissue slice pathology change:
Heart, liver, cerebral tissue to rat carry out sections observation, the results are shown in Figure 1.By Fig. 1-A, Fig. 1-B and Fig. 1-C as seen, in the aged control, the local special mess myocardial atrophy companion of heart small amount of fibers hamartoplasia, hepatocyte moderate steatosis around the liver central vein, a large amount of cavity sample aging variations appear in brain, illustrate that the variation of old and feeble property has all appearred in heart, liver, the cerebral tissue of aged control rat; And Mytilus Coruscus Polysaccharides is low, in high dose group and the young control, except that slight fat distortion appearred in the Mytilus Coruscus Polysaccharides low dose group, heart, liver, cerebral tissue were all normal, do not see special variation, thus the prompting Mytilus Coruscus Polysaccharides can delay the senescence process of important organ.
4.2.4 rat cerebral tissue's beta galactosidase assay
SABC is measured beta galactosidase and is seen Fig. 2 in corticocerebral distribution and content results.As seen from Figure 2, beta galactosidase is expressed more in the aged control rat cerebral cortex, and the expression in, high dose group low at Mytilus Coruscus Polysaccharides and the young control rat cerebral cortex all obviously reduces, the content of beta galactosidase is regarded as the relevant biomarkers of cerebral tissue aging usually, thereby the explanation Mytilus Coruscus Polysaccharides can delay the cerebral tissue senescence process.
4.2.5 the main biochemical indicator of serum is measured
The statistic analysis result that the blood glucose of serum (Glu), glutamate pyruvate transaminase (ALT), glutamic oxaloacetic transaminase, GOT (AST), blood urea nitrogen (BUN), inosine (CRE), cholesterol (CH) detect sees Table 2.
Table 2. respectively organize the main biochemical indicator of rat blood serum measurement result (x ± s, n=5)
*P<0.05
By table 2 as seen, Mytilus Coruscus Polysaccharides is low, high dose group is compared every index no difference of science of statistics with youth, aged control, illustrates that Mytilus Coruscus Polysaccharides is not in the mood for, liver, nephrotoxicity, and blood glucose is not had obvious influence yet.
4.2.6 SOD, MDA, GSH-px measure in the serum, the results are shown in Table 3.
By table 3 as seen:
(1) Mytilus Coruscus Polysaccharides high dose group SOD activity significantly increases (P＜0.01) than aged control, and the Mytilus Coruscus Polysaccharides low dose group is than aged control height (P＜0.05), and compare equal no difference of science of statistics with young control (P〉0.05);
(2) Mytilus Coruscus Polysaccharides high dose group GSH-px specific activity aged control significantly increases (P＜0.01), and the Mytilus Coruscus Polysaccharides low dose group is than aged control height (P＜0.05), and compares equal no difference of science of statistics with young control;
(3) Mytilus Coruscus Polysaccharides high dose group MDA content significantly is lower than aged control (P＜0.01), compares no difference of science of statistics with young control.
This shows that Mytilus Coruscus Polysaccharides has significantly anti-aging effects, antioxidation may be one of mechanism of its anti-aging effects.
Table 3. respectively organize rat blood serum SOD, MDA, GSH-PX enzyme activity measurement result (x ± s, n=5)
*P＜0.01 is with aged control;
*P＜0.05 is with aged control
Mytilus Coruscus Polysaccharides has significantly anti-aging effects to old SD rat under this dosage, antioxidation may be one of mechanism of its anti-aging effects.
4.2.7 SOD, MDA, GSH-px measure in the rat liver tissue, the results are shown in Table 4.
By table 4 as seen:
(1) Mytilus Coruscus Polysaccharides high dose group SOD activity obviously increases (P＜0.05) than aged control, and with young control no difference of science of statistics (P〉0.05) relatively;
(2) Mytilus Coruscus Polysaccharides high dose group GSH-px activity is compared with aged control, has significant difference (P＜0.01) and apparently higher than Mytilus Coruscus Polysaccharides low dose group (P＜0.05), and compares no difference of science of statistics with young control;
(3) Mytilus Coruscus Polysaccharides high dose group MDA content is lower than aged control (P＜0.05).
Table 4. respectively organize rat liver tissue SOD, MDA, GSH-PX enzyme activity measurement result (x ± s, n=5)
*P＜0.01 is with aged control;
*P＜0.05 is with aged control
This shows that Mytilus Coruscus Polysaccharides has significantly antioxidation, shows that it has anti-aging effects.
4.2.8 SOD, MDA measure in the rat cerebral tissue, the results are shown in Figure 3.
As seen from Figure 3:
(1) cerebral cortex SOD is active finds that Mytilus Coruscus Polysaccharides is low, the cerebral cortex SOD activity of high dose group rat is significantly higher than aged control (p＜0.01), and compares no difference of science of statistics with young control;
(2) Mytilus Coruscus Polysaccharides high dose group cerebral cortex MDA content significantly is lower than aged control (p＜0.01), with and the young control no difference of science of statistics; Show that Mytilus Coruscus Polysaccharides has significantly anti-aging effects, antioxidation may be one of mechanism of its anti-aging effects.
Above-mentioned zoopery result shows, Mytilus Coruscus Polysaccharides can improve the activity of SOD and GSH-px in serum, liver, the cerebral tissue respectively, reduces the content of MDA, so Mytilus Coruscus Polysaccharides has antidotal activity, can be used for preparing the medicine or the food of slow down aging.
Claims (2)
1. Mytilus Coruscus Polysaccharides is in the application of preparation antiaging agent or food.
2. the preparation method of the described Mytilus Coruscus Polysaccharides of claim 1, step is as follows:
1) preparation mussel extract:
Get the fresh mussel meat of East Sea Mytilus crassitesta Lischke, with high speed agitator it is stirred homogenate, add routinely and use little fire instead behind the water boil and boil 4-5h, filter to get filtrate, this is the mussel extracting solution;
2) preparation Mytilus Coruscus Polysaccharides:
The mussel extracting solution is concentrated the centrifugal precipitation of abandoning in back, the supernatant volume ratio is the sevage method gradation extraction of chloroform: n-butyl alcohol=4:1, lose protein precipitation until water, add the long-pending 90-95% ethanol of triploid again, stir room temperature standing over night, the centrifugal supernatant of abandoning, to precipitate lyophilization, get Mytilus Coruscus Polysaccharides.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106913587A (en) * | 2015-12-25 | 2017-07-04 | 无限极（中国）有限公司 | Application of the Jadeite Mussel water extract in the health food with anti-oxidant and neuroprotection |
| CN107227327A (en) * | 2017-07-07 | 2017-10-03 | 山东省药学科学院 | A kind of preparation method of mussel oligosaccharides and products thereof and application |
| CN109090620A (en) * | 2018-08-09 | 2018-12-28 | 山东省药学科学院 | A kind of application of mussel polysaccharide and its catabolite |
| CN111004294A (en) * | 2019-12-31 | 2020-04-14 | 山东省药学科学院 | Preparation method of mussel oligosaccharide single component MOS-1 |
-
2009
- 2009-01-05 CN CNA2009100448816A patent/CN101455676A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106913587A (en) * | 2015-12-25 | 2017-07-04 | 无限极（中国）有限公司 | Application of the Jadeite Mussel water extract in the health food with anti-oxidant and neuroprotection |
| CN107227327A (en) * | 2017-07-07 | 2017-10-03 | 山东省药学科学院 | A kind of preparation method of mussel oligosaccharides and products thereof and application |
| CN107227327B (en) * | 2017-07-07 | 2020-08-18 | 山东省药学科学院 | Preparation method of mussel oligosaccharide, and product and application thereof |
| CN109090620A (en) * | 2018-08-09 | 2018-12-28 | 山东省药学科学院 | A kind of application of mussel polysaccharide and its catabolite |
| CN109090620B (en) * | 2018-08-09 | 2022-02-22 | 山东省药学科学院 | Mussel polysaccharide and application of degradation product thereof |
| CN111004294A (en) * | 2019-12-31 | 2020-04-14 | 山东省药学科学院 | Preparation method of mussel oligosaccharide single component MOS-1 |
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Open date: 20090617 |