CN112716932A - Application of fucoxanthin in preparation of medicine or food for preventing or treating postoperative cognitive dysfunction - Google Patents

Application of fucoxanthin in preparation of medicine or food for preventing or treating postoperative cognitive dysfunction Download PDF

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CN112716932A
CN112716932A CN202110095199.0A CN202110095199A CN112716932A CN 112716932 A CN112716932 A CN 112716932A CN 202110095199 A CN202110095199 A CN 202110095199A CN 112716932 A CN112716932 A CN 112716932A
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fucoxanthin
cognitive dysfunction
mice
group
preventing
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董佳惠
崔巍
严小军
何山
张金荣
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Ningbo University
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Ningbo University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/336—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having three-membered rings, e.g. oxirane, fumagillin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Abstract

The invention relates to application of fucoxanthin in preparing a medicament or food for preventing or treating postoperative cognitive dysfunction. The medicine comprises one or more of tablets, capsules, pills, injections, oral agents and suspensions containing fucoxanthin or salts thereof. The food product comprises an edible body comprising fucoxanthin or an inner salt thereof. The invention provides an application of fucoxanthin in preparing a medicament or food for preventing or treating postoperative cognitive dysfunction, and the fucoxanthin has a good effect of preventing or treating postoperative cognitive dysfunction.

Description

Application of fucoxanthin in preparation of medicine or food for preventing or treating postoperative cognitive dysfunction
Technical Field
The invention relates to a new application of fucoxanthin, in particular to an application of fucoxanthin in preparing a medicament or food for preventing or treating postoperative cognitive dysfunction.
Background
Fucoxanthin (fucoxanthin) is also called fucoxanthin, fucoxanthin and fucoxanthin, and is widely distributed in algae of Phaeophyceae, Diatomidae, Chrysophyceae and dinoflagellaceae, and participates in photosynthesis. Fucoxanthin belongs to xanthophyll compounds of carotenoid family, has a special propylene carbon skeleton, has extremely strong oxidation resistance, can effectively eliminate free radicals and singlet oxygen, and prevents free radical damage.
Modern pharmacological research finds that fucoxanthin has good safety, and can have various pharmacological activities such as anti-tumor, anti-oxidation, anti-inflammation, weight reduction and the like in a human body. Such as: reports show that in the anti-tumor function, fucoxanthin can be applied to the prevention and treatment of skin cancer, colon cancer, blood system tumor, prostatic cancer and liver cancer; fucoxanthin can regulate Na+-K+ATP enzyme activity, which has good anti-oxidation effect, even better than vitamin E and vitamin C, especially has protection effect on human fiber cell injury caused by UV-B, and can regulate the activity of catalase and glutathione in tissues and molecules caused by retinol deficiency; in the anti-inflammatory function, fucoxanthin has an inhibiting effect on exudation of inflammatory mediators induced by endotoxin, and is in a dose-dependent relationship, the anti-inflammatory effect of the fucoxanthin is equivalent to that of prednisolone, and the fucoxanthin possibly has an important effect on various inflammatory reactions; in addition, fucoxanthin can eliminate fat accumulation in two ways, promote lipolysis, and stimulate liver to produce DHA for lowering cholesterol level.
Post-operative cognitive dysfunction (POCD) is a central nervous system complication occurring after an old person operation, and mainly shows that the cognitive function is damaged, such as impaired memory, mental concentration capability, language comprehension and lowered social adaptation capability, if the surgery is short, days to weeks, and if the surgery is long, months or even years. The patient death rate is increased, the rehabilitation is delayed, other complications are increased, the hospitalization time is prolonged, the medical expense is increased, and even a part of patients suffer permanent cognitive dysfunction, so that the life quality of the patients is seriously influenced. Therefore, there is significant clinical and social value in developing products that can combat post-operative cognitive dysfunction. The pathogenesis of POCD is currently unclear and it is generally believed that neuropathy and cognitive dysfunction are induced or exacerbated by various types of stress stimuli, including inhibition of the central cholinergic system, elevation of oxidative stress levels, release of inflammatory factors, and the like. The stress factors include physical factors (surgical operation, chemical factors (narcotics), biological factors (diet), social factors (mental stress), and the like, and relate to a plurality of system disorders such as central nervous system, endocrine system, immune system, and the like.
POCD is different from neuropsychiatric diseases such as degenerative diseases of the central nervous system, emphasizes the damage of specific conditions such as operation and anesthesia on a plurality of systems such as the central nervous system, and further induces cognitive impairment; the first factor affecting the degeneration of the nervous system is aging, which is manifested by progressive damage of neurons and chronic degeneration of nerve function.
Based on the above current situation, the present application has studied the use of fucoxanthin in the preparation of a medicament or food for the prevention or treatment of post-operative cognitive dysfunction.
Disclosure of Invention
In view of the above disadvantages in the prior art, the present invention provides an application of fucoxanthin in the preparation of a medicament or food for preventing or treating post-operative cognitive dysfunction.
The invention is solved by the following technical scheme.
Application of fucoxanthin in preparing medicine for preventing or treating postoperative cognitive dysfunction is provided.
Preferably, the medicament comprises one or more of tablets, capsules, pills, injections, oral preparations and suspensions containing the fucoxanthin or the salts thereof.
Use of fucoxanthin in the preparation of a food product for the prevention or treatment of post-operative cognitive dysfunction.
Preferably, the food product comprises an edible body comprising fucoxanthin or an inner salt thereof.
Compared with the prior art, the invention has the following beneficial effects: the invention provides an application of fucoxanthin in preparing a medicament or food for preventing or treating postoperative cognitive dysfunction, and the fucoxanthin has a good effect of preventing or treating postoperative cognitive dysfunction.
Drawings
FIG. 1 shows the recognition index of each group of mice in the training phase of the novel object recognition experiment according to the embodiment of the present invention.
FIG. 2 is a graph showing the recognition index of each group of mice in the exploration period of the novel object recognition experiment according to the embodiment of the present invention.
FIG. 3 shows the number of platform crossings for each group of mice in the water maze experiment according to the embodiment of the present invention.
FIG. 4 shows the target quadrant exploration time for each group of mice in the water maze experiment according to the embodiment of the present invention.
FIG. 5 shows the target arm search time of each group of mice in the Y maze experiment according to the embodiment of the present invention.
FIG. 6 shows the levels of antioxidant enzyme SOD in the brains of the mice in each group of the present invention.
FIG. 7 shows the levels of antioxidase CAT in the brain of mice in each group according to the example of the present invention.
FIG. 8 shows the expression of the protein related to AKT pathway in the brain of each group of mice in accordance with the present invention.
FIG. 9 shows the expression of ERK pathway-related proteins in the brain of mice in each group according to the present invention.
FIG. 10 shows the expression of inflammatory factor Tnf-alpha in the brain of each group of mice in the present example.
FIG. 11 shows the expression of IL-1 β, an inflammatory factor in the brain of mice in each group according to the present invention.
Description of the drawings:
FIG. 1: the mouse has normal recognition index and no obvious difference in the training period of the new object recognition experiment.
FIG. 2: administration of fucoxanthin or curcumin was effective against a reduction in recognition index (p <0.05) in POCD mice during the exploration phase of the novel object recognition experiment.
FIG. 3: the frequency of platform crossing in a water maze experiment of a POCD mouse is effectively improved by administering the fucoxanthin or the curcumin (p is less than 0.05).
FIG. 4: the curcumin administration effectively improves the exploration time of the POCD mice in a target quadrant of a water maze experiment (p is less than 0.01).
FIG. 5: the administration of fucoxanthin or curcumin effectively increased the in-target arm exploration time (p <0.05) in the Y maze experiment in POCD mice.
FIG. 6: the administration of fucoxanthin or curcumin effectively increased the levels of SOD in the brain of POCD mice (p < 0.01).
FIG. 7: the administration of fucoxanthin or curcumin effectively increased the levels of CAT in the brain of POCD mice (p < 0.01).
FIG. 8: administration of fucoxanthin inhibits the AKT pathway.
FIG. 9: inhibition of the ERK pathway by fucoxanthin administration
FIG. 10: curcumin or fucoxanthin administration was effective in reducing Tnf- α levels ([ P ] <0.05, [ P ] <0.01) in the brains of POCD mice.
FIG. 11: curcumin or fucoxanthin administration effectively reduced IL-1 β levels (/ P <0.05,/P <0.01) in the brains of POCD mice.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.
The invention relates to application of fucoxanthin in preparing a medicament for preventing or treating postoperative cognitive dysfunction, in particular to one or more of tablets, capsules, pills, injections, oral preparations and suspensions containing fucoxanthin or salts thereof.
Specifically, the structural formula of the fucoxanthin in the application is shown as follows.
Figure BDA0002913885780000041
Fucoxanthin can be extracted from herba Zosterae Marinae.
The first embodiment is as follows: fucoxanthin is effective in improving anesthesia and postoperative cognitive and memory disorders in mice caused by surgery, as follows.
1. Experimental animals: the mice are all purchased from the animal center of Zhejiang province, are healthy SPF grade ICR male mice, are 14 months old and have the weight of 40-45 g.
2. Experimental drugs: 480 mg of fucoxanthin, 6 mg of sodium carboxymethylcellulose and 30. mu.l of tween-20 were dissolved in 6 ml of physiological saline and mixed well in the dark. The total volume of curcumin 480 mg and sodium carboxymethylcellulose 24 mg dissolved in physiological saline is 6 ml, and the mixture is uniformly mixed in a dark place.
3. POCD animal model:
the POCD model of the aged mice refers to a plurality of literatures and is a typical model. The anesthetic was intraperitoneally injected 10 minutes before the operation (0.2 ml of both droperidol and fentanyl was diluted to 1 ml with physiological saline, and the anesthetic was prepared in a ratio of droperidol to fentanyl 1: 2 before use) 0.3 ml/40 g. When the mouse enters a shallow numb state (showing that the mouse stops crawling and then is in a side lying state, and part of the mouse has the angle arch rebound and has no limb action response to non-pain stimulation), the four limbs of the mouse are fixed by using the binding ropes and the mouse board, and the ventral surface of the mouse is upward. The hair of the mouse from the root of the hind limb to the lower edge of the thoracic cavity is removed, and a cotton swab is dipped with a proper amount of iodophor and smeared from the middle to the periphery. A2-3 cm wound is cut at the center of the ventral surface of the mouse, and the liver, the stomach, the spleen, the duodenum and the like of the mouse are sequentially turned over by using toothless forceps until the appendix and the omentum majus are explored. Avoid damaging the mesentery and blood vessel of the mouse. The small intestine, 5 cm of the same mesenteric artery loop donor area, was freed outside the abdominal incision for about 3 minutes. Suturing the peritoneal cavity and the skin, disinfecting the wound by using iodophor, and finally applying the erythromycin eye ointment to the wound.
4. The behavioral detection method comprises the following steps:
4.1 New object recognition
The experiment was completed in two days, the first day being the training period and the second day being the exploration period.
During the training period, the recognition index was calculated by first placing two identical objects diagonally in an open experimental box in the dark and recording the time each group of mice explored both objects within 5 minutes. During the exploration period, one of the objects was replaced with a new object of a different shape and the time to explore the new and old objects was recorded for each group of mice over 5 minutes and the recognition index was calculated.
4.2 Morris Water maze
The experiment is completed in five days, and the water temperature is kept at 20-25 ℃ in the experiment process. The mice were freely swimming for 90 seconds in a platform-free swimming pool on the first day, and were acclimatized. And enabling the platform to be exposed 0.5-1 cm above the water surface on the next day, so that each group of mice can easily find the platform and feel the direction of the platform. And immersing the platform in water for 0.5-1 cm for two days, and putting the mouse into the water maze from two quadrants and four quadrants to find the platform for one group. Two groups of experiments are carried out every day, namely, the mouse is trained for 4 times, the memory of the mouse on the platform position is continuously strengthened, and the movement track of the mouse and the time of reaching the platform are recorded through software. And removing the platform in the last day, and detecting the detection time of the mouse around the platform and the times of passing through the area where the platform is located.
4.3, Y maze
The experiment was completed in one day in a Y-maze with three arms equal in length and about 40 cm in height and uniformly laid with a layer of padding. Training period one arm was closed and mice were left to explore freely for 5 minutes in the other two arms. After 2 hours the arm was opened and the mice were allowed to explore from the same site for 3 minutes and the time to enter the newly opened target arm was recorded.
5. Experiment grouping
Selecting 16 mice with similar size and weight, randomly dividing the mice into 4 groups, wherein each group comprises 4 mice, and grouping the mice according to the mode shown in the table 1:
table 1 experimental grouping scheme
Figure BDA0002913885780000061
Figure BDA0002913885780000071
6. Results of the experiment
TABLE 2 identification indices (%) -of each group of mice in training and exploration phases in the New object identification experiment
Figure BDA0002913885780000072
From the data in table 2, the recognition indexes of each group of mice were averaged in the training period and the search period, respectively, and the data represent the average ± SD. The results are shown in FIGS. 1-2. As can be seen from fig. 1, in the new object identification experiment, the identification indexes of the mice in each group during the training period are not significantly different, which indicates that the factors such as the experimental environment and the used object do not affect the exploration condition of the mice. As can be seen from fig. 2, the recognition index during the exploration period between the post-operative cognitive dysfunction model group and the blank group is significantly different (p <0.01), indicating that the post-operative cognitive dysfunction mouse model is successfully established. In addition, the recognition index of the administration experimental group and the positive control group is obviously different compared with the cognitive dysfunction model after the operation (p < 0.05).
TABLE 3 platform crossing times for groups of mice in the Water maze experiment
Figure BDA0002913885780000073
TABLE 4 target quadrant exploration time(s) for each group of mice in the Water maze experiment
Figure BDA0002913885780000081
The number of platform crossings and target quadrant search time of each group of mice were averaged based on the data in tables 3 and 4, and the data represent the average ± SD, and the results are shown in fig. 3 to 4. As can be seen from fig. 3 and 4, the number of times of passing through the platform position in the post-operative cognitive dysfunction model group is significantly different (p <0.01) compared with that in the blank group, and the target quadrant exploration time is also different (p <0.05) from that in the control group, which indicates that the post-operative cognitive dysfunction mouse model is successfully established. In addition, the data of the administration experimental group are slightly higher than those of the postoperative cognitive dysfunction model group, and the data are close to those of the blank group. And the crossing times of the administration experimental group are obviously different from those of the postoperative cognitive dysfunction model group (p < 0.05). The crossing times of the positive control group and the exploration time of the target quadrant are all significantly different from those of the blank group.
TABLE 5 target arm exploration time(s) for each group of mice in the Y maze experiment
Figure BDA0002913885780000082
The target arm search time of each group of mice was averaged from the data in table 5, and the data represents the average ± SD, and the results are shown in fig. 5. As can be seen from fig. 5, the target arm exploration time of the postoperative cognitive dysfunction model group has a significant difference (p <0.01) compared with that of the blank group, which indicates that the establishment of the postoperative cognitive dysfunction mouse model is successful. In addition, the exploration time of the administration experimental group is significantly different (p <0.05) compared with that of the postoperative cognitive dysfunction model group.
The experimental results show that the fucoxanthin can effectively improve the mouse postoperation cognition and memory disorder caused by anesthesia and operation.
Example two: fucoxanthin counteracts post-operative cognitive dysfunction by reducing oxidative stress levels.
1. Experimental methods
Determination of antioxidant enzyme Activity
The antioxidant enzyme assay kit (Nanjing institute of biotechnology, Jiangsu) measures specific markers of oxidative stress, including SOD and CAT. According to the principle of enzymatic reaction, a reaction reagent is added and a blank is set. Measuring absorbance of SOD and CAT at wavelength of 450nm and 405nm, and calculating specific contents of SOD and CAT according to formula.
2. Results of the experiment
The fucoxanthin and the curcumin increase the antioxidant enzyme content in the brain of the mouse with the postoperative cognitive dysfunction.
Detecting the level of antioxidase (SOD and CAT), and evaluating the change of fucoxanthin to oxidative stress. As shown in fig. 6 and 7, the levels of SOD and CAT in the POCD model group were significantly reduced compared to the blank group. CAT and SOD levels were elevated in the dosed experimental and positive control groups (p <0.01, fig. 6, 7).
Example three: fucoxanthin reduces inflammatory factor release by inhibiting AKT and ERK pathways.
1. Experimental animals: are purchased from the animal center of Zhejiang province, are healthy male mice of the ICR with SPF grade, have the age of 14 months and the body weight of 40-45 g.
2. Experimental drugs: the fucoxanthin is divided into three concentrations, namely low concentration and medium concentration. And (3) low concentration: 120 mg, 1.5 mg of sodium carboxymethyl cellulose and 6 microliter of tween-20 are dissolved in physiological saline to total 6 milliliters; medium concentration: 240 mg, 3 mg of sodium carboxymethylcellulose and 15 microliter of tween-20 are dissolved in physiological saline to total 6 milliliters; high concentration: 480 mg, 6 mg of sodium carboxymethylcellulose and 30 microliters of tween-20 are dissolved in physiological saline to total 6 milliliters, and the components are uniformly mixed in dark. The curcumin 480 mg and the sodium carboxymethylcellulose 24 mg are dissolved in 6 ml of physiological saline in total, and are uniformly mixed in a dark place.
3. POCD animal model: the same as in the first embodiment.
4. Experiment grouping
Selecting mice with similar size and weight, randomly dividing the mice into 4 groups, wherein each group comprises 8 mice, and dividing the mice into groups according to the mode shown in the table 6:
TABLE 6 Experimental grouping protocol
Figure BDA0002913885780000101
5. Experimental methods
5.1 Western blot experiment
After 20 days from the start of the experiment, brain tissue of each group of mice in example 3 was collected, 10. mu.L of a lysis solution of RIPA-PIC-phosphatase inhibitor was added to 1 mg of the brain tissue, and the mixture was allowed to stand at room temperature for 1 hour, and then centrifuged at 13200rpm at 4 ℃ for 30 minutes. Taking a proper amount of supernatant, and carrying out the following steps: 1 amount of buffer 5 times added. And (3) uniformly mixing, heating in a water bath at the temperature of 99 ℃ for 5-10 minutes to denature the protein, and completing the preparation of the protein sample.
After adding the protein sample into the agarose gel, adjusting the voltage to 60V electrophoresis for 35 minutes, and adjusting the voltage to 80V electrophoresis for 100 minutes, so as to separate the proteins with different molecular weights. Then, the gel was taken out and covered with a cellulose acetate membrane, and 100V membrane transfer was performed for 90 minutes to transfer the proteins in the agarose gel to the cellulose acetate membrane. The cellulose acetate membrane was removed and then blocked with 5% skimmed milk powder solution at room temperature for 1 hour, after which primary antibody (formulated with 5% BSA) was added and incubated overnight, and then washed 6 times with 1 XTSST for 10 minutes each. After 1 hour incubation with the addition of secondary antibody, wash 6 times with 1 × TBST for 10 minutes each. And finally, adding color development liquid to the strips and placing the strips into an exposure machine for exposure. Checking the expression conditions of beta-actin, AKT, phosphorylation AKT and downstream protein GSK-3 beta, phosphorylation GSK-3 beta, ERK, phosphorylation ERK and downstream protein HO-1.
5.2 enzyme-linked immunosorbent assay (ELISA) assay
The ELISA kit is used for measuring the contents of TNF-alpha and IL-1 beta in brain tissues. Equal amounts of protein were loaded in all wells and the Optical Density (OD) was measured in an enzyme-labeled tester at a wavelength of 450nm and compared to a standard curve. All results were compared to the blank set of scenes.
6. Results of the experiment
Fucoxanthin reduces inflammatory factor release by inhibiting AKT and ERK pathways.
The expression conditions of beta-actin, AKT, phosphorylated AKT and downstream proteins GSK-3 beta, phosphorylated GSK-3 beta, ERK, phosphorylated ERK and downstream proteins HO-1 in the brain of the mouse are detected through a western blot experiment. The results show that compared with the POCD model group, fucoxanthin concentration dependently reduces AKT phosphorylation level and increases GSK-3 beta phosphorylation level; ERK phosphorylation decreased and HO-1 expression increased. Meanwhile, in an enzyme-linked immunosorbent assay, compared with a POCD model group, fucoxanthin concentration-dependently reduces the levels of inflammatory factors Tnf-alpha and IL-beta, which indicates that fucoxanthin can reduce the release of the inflammatory factors by inhibiting AKT and ERK pathways and is used for resisting postoperative cognitive dysfunction.
In the first embodiment, the administration mode is changed into intravenous injection, and the effect of the same level is also achieved; in the second example, the drug was mixed with liquid food and administered, and the effect was also obtained at the same level.
The scope of the present invention includes, but is not limited to, the above embodiments, and the present invention is defined by the appended claims, and any alterations, modifications, and improvements that may occur to those skilled in the art are all within the scope of the present invention.

Claims (4)

1. Application of fucoxanthin in preparing medicine for preventing or treating postoperative cognitive dysfunction is provided.
2. The use of fucoxanthin in the preparation of a medicament for preventing or treating post-operative cognitive dysfunction according to claim 1, wherein said medicament comprises one or more of a tablet, a capsule, a pill, an injection, an oral formulation, a suspension comprising fucoxanthin or a salt thereof.
3. Use of fucoxanthin in the preparation of a food product for the prevention or treatment of post-operative cognitive dysfunction.
4. Use of fucoxanthin in the preparation of a food product for the prevention or treatment of post-operative cognitive dysfunction according to claim 1, wherein said food product comprises an edible body comprising fucoxanthin or an inner salt thereof.
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WO2012065182A2 (en) * 2010-11-12 2012-05-18 University Of Massachusetts Modulation of ubiquitination of synaptic proteins for the treatment of neurodegenerative and psychiatric disorders
CN105412062A (en) * 2015-11-06 2016-03-23 宁波大学 New application of fucoxanthin to preparation of medicine for treating alzheimer disease
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