CN105106185A - Application of astaxanthin in preparation of medicine preventing and curing diabetic depression - Google Patents

Abstract

The invention discloses application of astaxanthin in the preparation of a pharmaceutical composition recovering a phosphorylated AKT level in diabetic depression prevention and treatment; the invention also discloses the pharmaceutical composition preventing and curing diabetic depression, and the pharmaceutical composition is characterized by containing the astaxanthin.

Description

The application of astaxanthin in the depressed medicine of preparation prophylactic treatment diabetes

Technical field

The present invention relates to astaxanthin and preparing the application in prevention and therapy diabetes depression, more particularly, the present invention relates to astaxanthin and preparing in prevention and therapy diabetes depression the application recovered in the pharmaceutical composition of phosphorylation AKT level.

Background technology

Depression is also known as depressive disorder, low for main clinical characteristics with remarkable and lasting mental state, is the main Types of mood disorders.Depression is a kind of common disease of psychiatric department, and it seriously perplexs the live and work of patient, brings serious consequence to society and family.Most of patients with depression tides over all one's life in fear, has the patient of many depressions to take one's own life in advance owing to being impatient at long-term spiritual misery and torment, according to the literature: the patients with depression of about 15% dies from suicide.Clinical visible mental state is low unbecoming with its situation, and the downhearted of emotion can from depressed to extremely grieved, and depression of feeling oneself inferior is even pessimistic and worldweary, can have suicidal attempt or behavior; Even occur numb; Some cases have obvious anxiety and mobility intense; The psychotic symptoms such as hallucination, vain hope can be there is in severe patient.Even each outbreak continues at least 2 weeks more than, elder's several years, majority of cases has the tendency of recurrent exerbation, and each outbreak great majority can be alleviated, and part can have residual symptoms or transfer to chronic.Depression is at present more and more for social concerns, and main because patient, except mood is low, oppressive, dejected, except despair, usually has the idea of suicide.According to statistics, the depressed patient of about 1/4 is dead because of suicide, and in crowd suicide, often 1/3 is depressed patient, and as can be seen here, Cure of depression patient is very important in time and energetically.Patients with depression will become the primary killers of the mankind in this century, and its harm is as follows: 1. emotion causes suicide and homicide; 2. emotion causes vehicle accident, death; 3. emotion causes industrial accident, death; 4. emotion causes collective's conflict, ethnic conflict, death.

World Health Organization (WHO) (WHO) tell us about the data of the investigation report of depression: depression rate is high, relates to crowd wide, and harm is serious.But only have the early discovery of few people energy, less people takes correct effective treatment, thus causes tragedy again and again to be showed, and creates the illusion of " depression=incurable disease ".Whole world patients with depression reaches 3.5 hundred million people, and depression annual loading is about 11%, about has 1,000,000 people therefore to commit suiside every year.The lifetime prevalence of China's depression reaches 3% to 7%, and patient numbers has reached 9,000 ten thousand people.The stimulation, physical disease etc. of the morbidity of depression and inherited genetic factors, characteristics of personality, adverse events all have relation, these factors can cause organism nervous system to get muddled, neurotransmitter secretion is unbalance, and brain electrical acti loses normal rhythm, thus causes the symptom appearance such as depressed.Therefore, only cannot produce therapeutical effect to depression with self regulation, must start with from root, by professional treatment means such as medicine, psychology, physics, could fundamentally Cure of depression.Depression is the disease that homicide rate is the highest.Depression has become first important diseases causing serious burden in global disease to the mankind, to the misery that patient and family members thereof cause, the loss that society is caused be other diseases incomparable.Cause the main cause of this situation be society correct understanding is lacked to depression, prejudice make patient be reluctant psychiatric department go to a doctor.In China, only have the patients with depression of 5% to accept treatment, a large amount of patients can not get diagnosing treatment timely, and sb.'s illness took a turn for the worse, even occurs the serious consequence of committing suiside.On the other hand, because the common people lack the knowledge about depression.Patients with depression is due to depressed, pessimistic and worldweary.Be easy to time serious produce suicidal thought.Further, because patient's thinking logic is normal, the success rate implementing to commit suiside is also higher.Suicide is one of the most dangerous symptom of depression.According to the study, the homicide rate of patients with depression is higher than population 20 times.May have over half in society suicide crowd is patients with depression.The suicide of some unknown cause may suffer from serious depression, before death only not by Timeliness coverage.To just generation during certain order of severity at disease progression owing to committing suiside.So find disease early, and early treatment, extremely important to the patient of depression.Do not wait patient to commit suiside, just expect that he may suffer from depression.Certainly, there singly do not have oneself to be unfavorable, also has very large harm to the relatives of surrounding.

Inside our traditional consciousness, think that general long more fat people's temper is all relatively good, belong to carefree and contented, but a nearest research shows, true really not so, contrary with our idea, obese people more easily suffers from this disease of depression, this makes people very puzzled, can so obesity cause depression to occur? the Canada Dai Weimeilei of McMaster University and his colleagues find, a kind of gene mutation relevant with obesity also declines 8% relevant with depression risk." this is first really relevant with depression gene." Mei Lei says.Obesity is also likely cause depression in fact, and FTO obtains intensive performance at brain.Mei Lei to together, proposes the hypothesis that this mutant gene can increase depressed risk these true pendulum.When Canadian two broad scale research demonstrate this conclusion, research group is surprised, so they have dissected again two researchs of Britain and Switzerland's research worker.Four researchs draw identical result: have this mutant copy that depressed risk will be made to reduce by 8% in genome; If there are two copies, the range of decrease doubles.In fact whether people obesity has this effect.The investigation contacted between a relevant fast food consumption and depression shows, the obesity that diet causes likely causes mouse to occur the behavior of doubtful depression.But, dietary habit is how to produce these impacts actually, still unclear at present.For the mankind, obesity really and depressive symptom have and necessarily contact.But it should be noted that there is not direct causal connection between the two.Some people suffering from obesity also may cause occurring depressive symptom because of other factors, such as economic situation, heredity, or other complication etc.Shire horse and Fulton provide higher fatty acid, the high heat food of 12 weeks continuously to the mouse of experimental group, finally make their mouse of body weight considerably beyond control group.Then, they have carried out the performance testing of 3 anxiety and depression aspects to these mouse.The investigation contacted between a relevant fast food consumption and depression shows, the obesity that diet causes likely causes mouse to occur the behavior of doubtful depression.But, dietary habit is how to produce these impacts actually, still unclear at present.For the mankind, obesity really and depressive symptom have and necessarily contact.But it should be noted that there is not direct causal connection between the two.Some people suffering from obesity also may cause occurring depressive symptom because of other factors, such as economic situation, heredity, or other complication etc.

At present, worldwide incidence of obesity increases year by year, and World Health Organization (WHO) has been located as a kind of important disease, and it has become public health problem important in world wide.National survey shows, and within 2002, China adult Overweight and obesity incidence rate is respectively 22.8% and 7.1%, rises 38.6% and 97% respectively compared with 1992.Overweight and obesity has had influence on 200,000,000 6 thousand ten thousand populations.The total cost of obesity and treating correlative diseases thereof is up to 10,000,000,000 RMB/year, and this is especially obvious in big and medium-sized cities.Within 2007, in the display of Beijing area sample survey results, Overweight and obesity incidence rate reaches 36.4% and 13.5% respectively in BJ Union Hospital.In the U.S., the crowd of 64.5% is in overweight, fat state, and the expense of preventing and treating obesity and related complication thereof increases sharply, and cost in 2003 reaches 75,000,000,000 dollars, and cost in 2006 then increases to 90,700,000,000 dollars, and within 2009, cost is up to 1,470 hundred million dollars.Due to increasing sharply of incidence of obesity, the growth and the spectrum of disease that cause metabolic disease change.Obesity not only has a strong impact on health of people and quality of life, also can cause hypertension (overweight people's sickness rate is 25% ~ 55%), diabetes (overweight people's sickness rate is 14% ~ 20%), coronary heart disease (overweight people's sickness rate is 10% ~ 15%), hyperlipemia (overweight people's sickness rate is 35% ~ 53%), sleep apnea (overweight people's sickness rate is 10% ~ 20%), depression (overweight people's sickness rate is 70% ~ 90%) etc., other also comprise the disease directly related with life-span and quality of life such as tumor, infertility, calculus.When particularly obesity reaches morbid obesity (BMI is more than 40), its mortality rate can present the curve sharply increased.Having only loses weight rapidly and effectively could reverse this curve.

Nowadays the whole world all research and development good effect and also toxic and side effects little, the natural hypoglycemic medicine even had no side effect and antidepressant drug, since the beginning of the sixties in last century, the living marine resources with huge potentiality to be exploited are the focuses that the world of medicine pays close attention to always, find the important goal that new living marine resources have now become researches on natural drugs.Astaxanthin (astaxanthin, ASX) full name is 3,3 '-dihydroxy-bata-carotene-4,4 '-one, is a kind of Red carotenoids, is extensively distributed in marine bacteria, algae, in shell-fish and Fish, many bibliographical information astaxanthins have been had to infect at antioxidation, anti-inflammatory, anticancer, anti-helicobacter pylori (HP) and demonstrate powerful effect in uvioresistant lamp process, and rare toxic and side effects.

Increasing result of study shows: astaxanthin, at protection islet beta cell function, promotes that insulin resistant is prevented and treated diabetes aspect and had suitable medical value.The people such as ZhangM adopt carbon tetrachloride to set up Liver Fibrosis Model, use 20 respectively, 40,80mg/kg dosage carries out astaxanthin treatment to rat, laboratory observation obviously can alleviate the pathological change of hepatic fibrosis to astaxanthin, also can suppress the expression of NF-kb and TGF-β, thus reduces the activation of hepatic stellate cell and the generation of extracellular matrix, especially use heavy dose of astaxanthin intervention, it is larger that hepatic fibrosis improves degree.BhuvaneswariS finds that the non-oxidizability of astaxanthin and antiinflammatory action effectively can reduce the generation of hepatocyte endoplasmic reticulum oxidative stress, stops and delays the chronic sympton that inflammation causes.

Diabetes depressive illness is because of complexity, and development and insulin resistant occur for it, and fat metabolic disturbance is fat, oxidative stress and closely related, but the molecular mechanism that development occurs for relation between it and diabetes depression is still unclear.Clinically, first preventing and correct metabolism disorder of blood lipid, is to the primary prevention of diabetes or early treatment has positive effect.During treating diabetes, will pay close attention to mental status and change while controlling blood glucose, depression is recoverability damage in early days, and diagnosis and treatment early often can make depressed reverse, and these have great importance to prevention and therapy diabetes and complication thereof.

In prior art, also there is not medicine for recovering AKT phosphorylation level in depression.The present inventor sets up classical type 2 diabetes mellitus animal model, observe astaxanthin before high fat feeds STZ administration with become mould after to type 2 diabetes mellitus rat blood sugar, blood fat and fat impact, and astaxanthin to type 2 diabetes mellitus rat mental symptom make moderate progress effect time, surprisingly find astaxanthin can recover diabetes depressed time cerebral tissue in the level of phosphorylation AKT, thus complete the present invention.

Astaxanthin (astaxanthin) involved in the present invention, has another name called astaxanthin, Astaxanthin, is a Carotenoids, and be also the highest level product of carotenogenesis, in rediance, chemical constitution is similar to beta-carotene.And beta-carotene, phylloxanthin, canthaxanthin, lycopene etc. are all the intermediate products of carotenogenesis, therefore at nature, astaxanthin has the strongest non-oxidizability.Extensively be present in biosphere, in the feather of particularly shrimp, Eriocheir sinensis, fish, frond, yeast and birds, content is higher, is one of carotenoid main in marine organism.

Astaxanthin chemical name: 3,3 '-dihydroxy-4,4 '-diketo beta-carotene, pigment Aj067-69CASNo:472-61-7, molecular formula C40H52O4, molecular weight is 596.86.Due to hydroxyl (-OH) the optical activity reason at two ends, astaxanthin has 3S-3 ' S, 3R-3 ' S, these 3 kinds of isomery kenels of 3R-3 ' R (also referred to as left-handed, meso, dextrorotation), wherein synthetic astaxanthin is that the mixture of 3 kinds of structure astaxanthins (left-handedly accounts for 25%, dextrorotation accounts for 25%, meso about 50%), few antioxidant activity, completely different with the astaxanthin (based on transconfiguration---3S-3S type) in the aquaculture organism body such as salmon. the astaxanthin in yeast source is 100% dextrorotation (3R-3 ' R), has Partial Antioxidation active; Above-mentioned two kinds of source astaxanthins be mainly used in non-edible animal and goods and materials painted on.Only have the astaxanthin in algae source to be 100% left-handed (3S-3 ' S) structure, there is the strongest biologic activity.

Summary of the invention

Accompanying drawing explanation

Fig. 1: the experiment flow figure of the embodiment of the present invention 1.

Fig. 2: the experiment flow figure of the embodiment of the present invention 2.

Fig. 3: in the embodiment of the present invention 1, the change of each group rat blood sugar (is compared with matched group, * P < 0.05; Compare with high fat group, #P < 0.05).

Fig. 4: in the embodiment of the present invention 2, the change of each group rat blood sugar (is compared with matched group, * P < 0.05; Compare with diabetic groups, #P < 0.05).

Fig. 5: the change of each group rat OGTT area under curve in the embodiment of the present invention 1 and 2.

Fig. 6: each group rats underwent open field test (comparing with matched group, * P < 0.05, * * P < 0.01, #P > 0.05) in the embodiment of the present invention 1 and 2.

Fig. 7: in the embodiment of the present invention 1 and 2, each group rats underwent spontaneous activity test (is compared with matched group, * P < 0.05; #P > 0.05).

The change of Fig. 8: the embodiment of the present invention 1 and 2 Zhong Gezu rat cerebral tissue phosphorylation AKT level.

Detailed description of the invention

Materials and methods

1.1 laboratory animal

Healthy male Sprague-Dawley (SD) rat, in one month age of Mus, body weight 100-120g, is provided by Zhejiang Province's Experimental Animal Center.Animal sub-cage rearing, laboratory animal starts experiment again after animal center conforms one week, and free drinking water standard rat feed is fed, and keep clean in cage, room temperature controls at 24 DEG C, lamp, well-ventilated, and relative humidity is 60%.Zoopery completes in University Of Ningbo's Experimental Animal Center.

1.2 experimental apparatus and reagent

1.2.1 major experimental instrument

Table 1 experimental apparatus title and manufacturer

1.2.2 experimental drug and reagent

Table 2 experimental drug title and manufacturer

Experimental technique

1.2.32 the successful standard of patients with type Ⅰ DM model

Experimental rat first feed with high lipid food (in every 100g, each component ratio is for crude protein: fat: carbohydrate=20: 40: 40) 6 weeks, inject 1% streptozotocin (STZ) citrate buffer solution (being the 0.1mol/L sodium citrate buffer solution Fresh of 4.2-4.4 with pH) by the dosage disposable celiac of 25mg/kg again and carry out modeling, cut tail and get blood survey random blood sugar, random blood sugar value>=11.1mmol/L is the successful standard of type 2 diabetes mellitus model ³⁰.Observe the mental status of rat every day, ensure sufficient food and drinking-water.

1.2.4 pharmaceutical intervention

Prevention group administration time and treatment group administration time are respectively 4 weeks and 6 weeks, and given low is 25mg/kg, and all the other groups synchronously give the normal saline of equivalent, free diet drinking-water.

1.2.5 random blood sugar measures and oral glucose tolerance test

In chemoprophylaxis and therapeutic process, each group every rat surveyed a random blood sugar every 3 days at synchronization, gets hematometry, at once carry out relative recording by automatic blood glucose meter from rat tail vein, added up blood glucose value change after experiment terminates.

In zoopery is carried out, each group rat is respectively at pharmaceutical intervention 4 weeks rear row oral glucose tolerance tests.Concrete grammar is: after Rat Fast 12h, carries out gavage (given low is 2g/kg) with the solution of 50% glucose.After gavage 0h, 0.5h, 1h and 2h, cut tail get tail vein survey blood glucose value.Experiment terminates rear drafting each group of rat glucose tolerance curve, and area under calculating glucose tolerance curve.

1.2.6 blood sample collection

Blood collection: after treatment terminates, every rat selects angular vein to get blood, and concrete grammar is: get vacuum test tube ready in advance and carry out labelling, and left hand fixes rat, the glass capillary of right hand diameter 1mm about length 5cm is rotatably inserted into inside rat eye, adjust degree of depth posterior vein blood to flow out immediately, access blood with blood taking tube, leave standstill after 2-3h until blood, with the centrifugal 2000rpm of low speed centrifuge, 10min, gets supernatant, detects blood lipids index with automatic biochemical instrument.

1.2.7 open field test: also known as Open field test is a kind of method of evaluation experimental animal inner directed behavior, exploratory behavior and tensity in strange environment.Spacious field analysis system is the change of observational study laboratory animal psychoneural, enters the various actions after free environments, such as animal is mainly movable in neighboring area to the fear of new free environments, less in middle section activity, but the characteristic of probing into of animal impels again it to produce in the motivation of middle section activity, the also anxious psychology that produces therefrom of observable.Central stimulant can obviously increase autonomous activity and reduce exploratory behavior, and the antipsychotic drug of doses can reduce exploratory behavior and not affect autonomic activities.With the occurrence frequency of laboratory animal some behavior among novel environment and persistent period etc., the inner directed behavior of reaction experiment animal in foreign environment and exploratory behavior, react its tensity with urine and stool number of times.

1.2.8 spontaneous activity test: be the change of observational study laboratory animal psychoneural, enter the various actions after free environments, such as animal is mainly movable in neighboring area to the fear of new free environments, less in middle section activity, but the characteristic of probing into of animal impels again it to produce in the motivation of middle section activity, the also anxious psychology that produces therefrom of observable.Central stimulant can obviously increase autonomous activity and reduce exploratory behavior, and the antipsychotic drug of doses can reduce exploratory behavior and not affect autonomic activities.It is made up of experimental box (large mice), video camera, camera lens, data wire and camera power supply line, camera mount, PCI video frequency collection card, usb data softdog, the mounting disc of spontaneous experiment video analytic system.

1.2.9 forced swimming test: the research being mainly used in antidepressant, calmness and analgesic type drug.This system is applicable to rat, mice or other laboratory animals, by laboratory animal being placed in the environment (as water) of a limitation, animal in this context risk one's life struggle make an attempt at escaping and cannot escape, thus provide one without avoidable compressing environment, after the experiment of a period of time, namely animal shows typically " motionless state ", reflect one to be referred to as " behavioral despair state ", record is in the series of parameters in the motionless state procedure of the animal generation despair of this environment.This system uses photographic head to experimentation video recording and carries out trace analysis with corresponding software to image, and reduce manual operation, data are objective.The animal models of depression of forced swimming, be research human depression pharmacology and pathogeny thereof, screening to observe in antidepressant drug research reliably experimental model, its main feature is pharmaceutically-active high degree of specificity, this experiment can be good at antidepressant drug and strong stabilizing to be distinguished with antianxiety drugs, and the effect that produces of most of antidepressants and clinical potency significant correlation, this is by numerous scholars are accepted.

1.2.10Westernblot the change of an experimental group rat cerebral tissue phosphorylation AKT level is analyzed.

[embodiment 1]

Astaxanthin is to the Primary preventive intervention effect of type 2 diabetes mellitus depression

Experiment flow figure in embodiment 1 that Fig. 1 is real.Specifically, 20 healthy male SD rats are divided into five experimental grouies at random, be respectively: A group (Normal group), B group (simple high fat group), C group (high fat+STZ group), D group (high fat+astaxanthin prevention+STZ group), E group (high fat+astaxanthin prevention group), often organizes each 4 rats.Normal group is fed with normal rats feedstuff 10 weeks, all the other are respectively organized rat and all synchronously feed with high lipid food, and from the 7th week, D group and E group rat all prevented with the dosage astaxanthin of 25mg/kg, A, B, C group gives the normal saline of equivalent simultaneously, and the time cycle is 4 weeks.Prevention terminates to carry out empty stomach 12h to often organizing rat afterwards, next day C group and D group rat inject 1%STZ citrate buffer solution with the dosage disposable celiac of 25mg/kg, A, B, E group rat synchronously injects the normal saline of equivalent, and then row OGTT tests.Each group of rat glucose tolerance curve is described to show the change of rat glucose tolerance after testing and terminating, calculate and area value (AUC) under respectively organizing rat glucose tolerance curve, carry out each Behavior Test, finally put to death rat and collect serum and tissue samples, detect each group of rat blood serum biochemical indicator subsequently and carry out H & E and dye, observe each group of morphologic change of liver tissues of rats, and detect the change of phosphorylation AKT level in cerebral tissue by quantitative Westernblot method.

[embodiment 2]

Astaxanthin is to the therapeutical effect of type 2 diabetes mellitus depression

This experiment and above-mentioned preventive effect experimental synchronous carry out.As tested shown in flow chart (see Fig. 2): get male and healthy SD rat 20, first random selecting 4, only as Normal group, is fed with normal rats feedstuff 6 weeks.All the other group rats synchronously feed injects 1%STZ citrate buffer solution with the dosage disposable celiac of 25mg/kg with high lipid food after 6 weeks.Cut tail subsequently and get blood survey random blood sugar value, the animal choosing 8 random blood sugar value >=11.1mol/L is experimental rat, and is divided into diabetic groups and each 4 rats for the treatment of group at random.The dosage that treatment group gives astaxanthin is 25mg/kg; All the other groups give the normal saline of equivalent simultaneously, each Mus gavage every day 1 time, and the time cycle is 6 weeks, free diet drinking-water.After end to be treated, each group of rat carries out OGTT test, then each Behavior Test is carried out, rat is put to death subsequently, get rat blood serum and win cerebral tissue sample, carry out follow-up blood glucose value mutation analysis, extract fresh brain tissue histone, by Westernblot method in the change detecting phosphorylation AKT level in cerebral tissue.

Experiment shows, under diabetic conditions, experimental mouse open field test behavior frequency obviously reduces; Spontaneous activity test motion frequency obviously declines.After astaxanthin treatment, above-mentioned test value all returns to normal level.If give preventative process astaxanthin, then recovery even more ideal (as Fig. 6, Fig. 7 show), shows that astaxanthin has good preventive and therapeutic action to the depression that diabetes cause.Westernblot analyzes display, phosphorylation AKT level in experimental rat cerebral tissue is obvious decline in diabetes, if give astaxanthin prophylactic treatment before molding, higher level can be maintained when diabetes occur, if but do not give prophylactic treatment, then there is rear astaxanthin to the recovery effects of phosphorylation AKT level not good (as shown in Figure 8) in diabetes.

Table 4 description Chinese and English initialism synopsis

STZ	Streptozotocin
		AUC	Area under glucose tolerance curve
OGTT	Oral glucose tolerance test

Claims (10)

1. astaxanthin is preparing in prevention and therapy diabetes depression the application recovered in the pharmaceutical composition of phosphorylation AKT level.

2. apply as claimed in claim 1, wherein, described astaxanthin is natural astaxanthin.

3. apply as claimed in claim 1 or 2, wherein, described depression be selected from diabetes depression, Obesity depression extremely, one or more in constitutional depression.

4. apply as claimed in claim 1 or 2, wherein, described diabetes depression is caused by diabetic nephropathy.

5. apply as claimed in claim 1 or 2, wherein, described diabetes are depressed from type 2 diabetes mellitus rat.

6. a pharmaceutical composition for prevention and therapy diabetes depression, is characterized in that, containing astaxanthin.

7. pharmaceutical composition as claimed in claim 7, wherein, described astaxanthin is natural astaxanthin.

8. as claimed in claims 6 or 7 to go compositions, wherein, described depression be selected from diabetes depression, Obesity depression extremely, one or more in constitutional depression.

9. as claimed in claims 6 or 7 to go compositions, wherein, described depression is caused by type 2 diabetes mellitus.

10. as claimed in claims 6 or 7 to go compositions, wherein, described diabetes are depressed from type 2 diabetes mellitus rat.