CN101244160A - New anti-stress and tidepressant purpose of banana tegument extract - Google Patents
New anti-stress and tidepressant purpose of banana tegument extract Download PDFInfo
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Abstract
The invention relates to a medicine, a healthy food and a food additive, which adopt the extract of banana skin as raw material and is used for preparing anti-stress and antidepressant medicines and medicines for alleviating and improving the adverse reactions of hepatic injury caused by stress load. The medicine is based on that the extract of banana skin has regulation effect on the restraint stress mice plasma, anti-oxidation capacity of liver and the content of malon dialdehyde (MDA) in liver, and has significant effect on the improvement effect of increase of pyruvic transaminase of restraint stress mice plasma and on the in vitro anti-oxidation experiment. The usage amount of the extraction of banana skin in various finished products of medicine, healthy food and food additive can be 0.01 to 100% in weight ratio due to high safety and exact biological activity.
Description
Technical field
The present invention relates to field of medicaments, specifically is natural plant extracts and application thereof.
Background technology
Current social develop rapidly, people's quality of life improves year by year.But the problem that can not ignore is that in this course, the mental pressure from different aspect and level that people afforded is also increasing thereupon.In the face of many-sided stress such as nervous work, lives, being in the sub-health state crowd increases year by year, even the various focuses of holing up in the body, and this situation has become society's one big potential crisis in modern age.Various psychology and the physical stress that people bore on science, can be summarised as " stress ".Notion that stress (stress) proposed (Nature by Canadian scientist Selye in 1936,138:32-33,1936), and its stress is described as general adaptation syndrome (GAS) (general adaptationsyndrome), emphasize that simultaneously stress comprises many-sides such as spirit, nerve, endocrine and immunity, the concerted reaction of many tracts, the single comprehensive response status of right and wrong.Up to now Yan Jiu Certificate is bright, and to form disease from loading to be a kind of maincenter that comprises, endocrine and immune system are complicated in endosome stress the load response process.Stress be except that battle-weary cause, also exist the pathologic stress.Chronic fatigue syndrome (chronicfatigue syndrome) for example, the fatigue symptom of some diseases due to habit disturbance cause institute secondary such as diabetes, or the clinical symptoms that causes indirectly because of reasons such as drug side effectes, we can say that stress belongs to a kind of syndrome that loses homeostasis state in the body.In addition, stressor is defined as various factors that living individual is threatened again.These factors can be summarized as psychological factor, the factor (Neuroendocrinol, 20:1～48,1999) of physical factor and the dynamic equilibrium of change cardiovascular system.The result of stress is influenced by character, intensity and the persistent period of stressor and reactivity, the sensitivity of body.Stress load is a kind of common factor of causing a disease, cause injury.Especially chronic persistence stress be loaded and often be caused the pathologic damage of internal organs, tissue, causes metabolism disorder, brings out multiple disease, serious harm people's physical and mental health.
The variation of central neurotransmitter level is the important behaviour of stress, also is the basis (BiolPsych-iatry, 46:1167～1180,1999) that stress load and bring out psychology and behavior change.Because based on neurotransmitter and other informational molecules, therefore stress loading, central nervous system's functional activity can cause that neurotransmitter levels changes.Sudha people such as (Physiol Behav, 57:1061～1066,1995) finds to restrain and stress reduce DA level in rat prefrontal cortex and the striatum.Stress stimulations such as forced movement can increase the activity of tryptophan hydroxylase, increase the synthetic and metabolism (Brain Res, 901:247～251,2001) of 5-HT.
The experimental results proof stress be loaded and can be produced behavior disorder, and stimulates the prolongation of time-histories with stressor, and its behavior shows as by excited, anxiety state and transfers inhibition, depressive state to.Under stress loading strongly or for a long time, the body homeostasis is unbalance, the bad variation of tangible physiological change such as neuroendocrine imbalance, hypoimmunity, irritability gastrointestinal disease even the emotion and the mental status can occur, and severe patient can cause depressive symptom.
Though in Chinese medicine, do not have " stress " speech, people in secular practical experience of life, early have recognized that stress load and disease between relation.One of important content in tcm causa morbi-pathology, seven emotions cause a disease opinion be exactly about spirit stress with the most representative argumentation of disease relationship.Chinese medicine is emphasized " healthy energy deposit in, heresy can not be done ", and wherein healthy energy promptly refers to the normal neural ergasia of body, if the activity of feelings will is unusual, then can has influence on the function of internal organs and QI and blood and produces pathological changes, be i.e. the traditional Chinese medical science so-called " internal injury caused by excess of seven emotions ".In " interior warp ", also have a lot about the argumentation relevant of feelings will, said as " element is asked and lifted the pain opinion ": " negative and positive are born in gas also " with internal organs.The traditional Chinese medical science also has the theory of " excessive anger impairing the liver ": in internal organs, liver is had the greatest impact by feelings will, and because of liver controlling conveyance and dispersion, the happiness bar reaches dislikes depression, and the effect with harmonizing the functional activities of vital QI, feelings will is so feelings will is hindered mostly and liver has confidential relation.The traditional Chinese medical science often adopts the method for soothing the liver nourishing the liver to carry out auxiliary treatment when unusual when the activity of feelings will takes place.
Modern medicine study confirms that also emotionality stress obviously influence the functions such as nerve, endocrine and immunity of body, stress load or the generation development of the destruction of psychological defense mechanism and multiple disease has confidential relation.In neural, endocrine and this three's of immune system mutual relation, active oxygen (reactive oxygen species, ROS) or perhaps the damaging oxidation of oxygen-derived free radicals to attack be common basic mechanism in inherence between the three.Under the normal condition, the oxygen of body comes from the breathing of pulmonary, is brought into each histoorgan of whole body by the alveolar exchange by blood and supplies biogenic oxygen.Metabolic process also produces certain oxygen-derived free radicals simultaneously, and the strong oxidability of living radical has the noxious bacteria of killing under normal condition, and control is infected, the effect of environment in keeping.When body when stressor is loaded, the hypothalmus-pituitary-adrenal axis irritability improves, acth secretion glucocorticoid level raises, brain Vigilance improves, and kinetism improves, the blood glucose increased blood pressure, Pulse Rate increases, thereby mobilizes energy storage, conforms.At this moment, the blood of body flows in the peripheral circulation in a large number, makes internal organs be in a kind of state of hypoxic-ischemic.When the external world stress load releasing or alleviation, though the every physiological function of body is recovered, the ischemia-reperfusion of internal organs can cause the reactive oxygen free radical generation to be increased.Active oxygen is at the intravital meeting damage dna chain of accumulating of machine, and the infringement cell membrane promotes the senilism process, causes histoorgan generation pathological changes.
Liver is one of organ that blood flow enriches the most in the human body, and is very responsive to hypoxic-ischemic, is the main target that various active oxygens are attacked, and the toxic action of oxygen-derived free radicals plays key effect in the hepatic injury pathogenic process.Up to now experimental results show that liver acute ischemia again under the stress state of perfusion oxygen-derived free radicals increase unusually.The oxygen-derived free radicals that body produces by enzyme system and non-enzyme system, can attack polyunsaturated fatty acid (the polyunsaturated fatty acid in the liver biomembrane, PUFA) cause lipid peroxidation, generate lipid peroxidation product as malonaldehyde, ketone group, hydroxyl, hydroperoxy and oxygen-derived free radicals etc.Lipid peroxidation damages liver plasma membrane by chain reaction, produces the chain reaction that a large amount of new active oxy groups inspire lipid peroxidation simultaneously.Therefore the lipid peroxide that produces continues the liver injury cell, so one of main mechanism the when lipid peroxidation of liver plasma membrane is hepatocyte injury.Thereby the test lipid peroxide-MDA content can reflect the snperoxiaized degree of liver inner lipid, reflect the degree of liver cell damaged indirectly.Oxygen-derived free radicals causes that the result of lipid peroxidation descends the flowability of liver plasma membrane, and permeability increases, and glutamate pyruvate transaminase is released into blood, and the glutamate pyruvate transaminase vigor increases in the blood.Thereby the vigor of measuring glutamate pyruvate transaminase in blood plasma or the serum can be used as an important indicator of hepatic injury diagnosis.
In view of developing rapidly of the goods and materials civilization of modern society, people constantly accept increasing challenge, are in increasing various stress the load, and body descends to the defense function of active oxygen, and internal organs are in sub-health state for a long time.Therefore,, have anti-stress from healthy and living habit angle, antidepressant effect, can improve and alleviate stress load medicine, food and the food additive of the various untoward reaction such as hepatic injury that cause becomes the people's attention focus and presses for.
Anti-stress medicine can weaken the effect that stress load to body, reduce the reactivity of body, neuroleptanalgesiss agent (chlorpromazine, Lacumin, trifluoperazine, haloperidol), tranquilizer (chlorine dihydro aminomethyl phenyl benzodiazepine ketone, phenazepam) and tranquilizer (Benactyzire, sodium bromide, dibasole) are arranged usually loading.But these medicine majorities have side effect and are subjected to certain limitation on clinical using dosage, also on effect are confined to temporary alleviation more, and can not fundamentally reach therapeutic purposes.We are being devoted to seek and develop new anti-stress for this reason, in the process of antidepressant medicine, health food, food and food additive, also notice the anti-stress of Pericarpium Musae, and the antidepressant effect effect reaches its application and development.
Fructus Musae belongs to Musaceae (Musaceae) on Plant Taxonomy, the fruit of Musa (Musa) plant Fructus Musae Musa sapientum Linn..Fructus Musae originate in the south, Asia comprise southern china, India's Himalayas the foot of a mountain to the West Indies etc. the torrid zone local.The torrid zone, the general at present cloth world, subtropical zone are to cultivate tropical fruit (tree) the most widely in the world to the area, temperate zone.Be rich in compositions such as flavonoid, aminoacid, fatty acid, polysaccharide, trace element and vitamin A, B, C, E in the Fructus Musae, also contain materials (Science, 127:684-750,1958.) such as a small amount of norepinephrine, 5-hydroxy tryptamine and dihydroxy benzenes ethamine.In China, the medicinal beginning of Fructus Musae is stated from " the southern regions of the Yunnan Province book on Chinese herbal medicine ", and the sweet acid of its nature and flavor has the spleen invigorating removing food stagnancy, promoting the production of body fluid to quench thirst, heat-clearing and toxic substances removing, the effect of promoting blood circulation and hemostasis.Modern pharmacology studies show that Fructus Musae can leukocyte increasing, improves functions of immune system, shows the antioxidation with free radicals such as removing ROO and OH.Can obviously reduce the levels of peroxide of blood plasma low density lipoprotein, LDL (LDL) in vivo.Fructus Musae can strengthen the human plasma oxidation resistance, can play the positive role of angiocardiopathy preventing and slow down aging.Fructus Musae also can improve the concentration of 5-hydroxy tryptamine in the brain in addition, has effects such as the depressive symptom of improvement.
In recent years, people are concerned about the research to the natural product physiologically active very much, and are noticeable especially to traditional Chinese herbal medicine and biological function that natural plant kind has.The use history in existing thousands of years of tradition Chinese herbal medicine, these traditional Chinese herbal medicine to all regulatory functions of health since ancient times early with known.For example Fructus Musae can improve melancholy symptom, has certain anti-inflammatory effect, antibacterial action etc. and is familiar with from the life experience by people.But lack the evaluation of objective and science.Particularly for the anti-stress of Pericarpium Musae, the discussion that the untoward reaction such as hepatic injury that cause by loading still do not have any application facet is alleviated and improved to antidepressant.Therefore we are from the angle of the process of inducing an illness of stress loading, cause a disease theoretical in conjunction with the seven emotions of the traditional Chinese medical science, brake as stressor with constraint, duplicate mice stress model, inquire into the regulating action influence of Fructus Musae extract, observe the antioxidation in vitro effect that improve effect and Fructus Musae extract of Fructus Musae extract simultaneously Restraint Stress load mice plasma Glutamate pyruvate transaminase rises to mda content in Restraint Stress load mice plasma and liver oxidation resistance and the liver.
Summary of the invention
Goal of the invention of the present invention is to provide the new purposes of pericarp of Fructus Musae extract.
Of the present invention is raw material with the pericarp of Fructus Musae extract, is used to prepare anti-stress, and antidepressant is alleviated and improved in response to swashing the fatigue that load causes and the medicine of hepatic injury and clinical symptoms thereof.Drug prepared can be taken for a long time, and is safe and have no side effect.
1, pericarp of Fructus Musae extract of the present invention is a raw material, is used to prepare prevention and improves food and food additive by the fatigue that stress cause and hepatic injury and clinical symptoms thereof.Prepared food and food additive can be taken for a long time, and be safe and have no side effect.In its application product, the use amount of described pericarp of Fructus Musae extract can account for 0.001～100% weight ratio.
Pericarp of Fructus Musae extract of the present invention is to adopt one or more any mixed solvent of lower alcohols such as water or water and methanol, ethanol, acetone isopolarity solvent to extract.Because be rich in flavone, polyatomic phenol material and class nutritional labelings such as rich in amino acid, fatty acid, polysaccharide and trace element in the Pericarpium Musae, high polar compound in all belonging to, the lower alcohols of general water or variable concentrations, acetone isopolarity solvent or any mixed solvent extract.When extracting, can suit to adjust the response rate that determining alcohol is used for improving the Pericarpium Musae active component.When extracting, can influence the content of active component in the extract because of the change of conditions such as temperature, method and time.But for extraction of active ingredients in the Pericarpium Musae, said extracted method and combination in any thereof all can reach the recovery of extract of the present invention.
In view of pericarp of Fructus Musae extract is to develop as final products with medicine or health food and food additive, should select aquiferous ethanol to be advisable so consider to extract solvent from safety perspective as far as possible.The ratio of Pericarpium Musae and solvent does not have specific limited during extraction, but considers that generally the ratio of Pericarpium Musae and solvent is advisable with 1: 1～50 scopes.The present invention does not have specific limited to extracting temperature, and is all applicable to the solvent boiling point temperature from room temperature.Certainly room temperature, normal pressure and in the solvent boiling point scope, extract ideal.Extraction time can reach the recovery purpose of active component to a certain extent from 30 seconds to 24 hours.When extracting pericarp of Fructus Musae extract, also can in solvent, add materials such as sodium bicarbonate, SODIUM ASCORBATE as required.These extracting method can not influence the biological activity that Pericarpium Musae had originally.
Pericarp of Fructus Musae extract solid agent use amount provided by the invention is generally at 0.001% to 100% (W/W), and peroral dosage form interpolation concentration is generally comparatively suitable in 0.1% to 50% (W/W) scope usually.In this concentration range, pericarp of Fructus Musae extract is suitable for directly taking, and is accepted by consumer easily.Certainly, as the increase along with addition of the pericarp of Fructus Musae extract of food additive, pained sensation may appear.Its solution can require according to the psychology of market demand and consumer to change dosage form, for example uses dosage forms such as tablet, granule, capsule.Low concentration pericarp of Fructus Musae extract solution can be used as daily beverage and offers consumer.
Pericarp of Fructus Musae extract provided by the invention and Pericarpium Musae active component can be used for various peroral dosage forms and put on market, are used to prevent and improve the depression of loading and causing in response to swashing, tired and hepatic injury and clinical symptoms thereof.Certainly, pericarp of Fructus Musae extract also can combine with pharmaceutical carriers such as excipient commonly used and make preparation.In case of necessity, can also add preparation additives such as some antiseptic, antioxidant, pigment, sweeting agent.Relatively Shi Yi Pericarpium Musae excipient generally considers to have lactose, sucrose, mannitol, starch, crystalline cellulose etc.Lubricant generally should adopt magnesium stearate, calcium stearate, Pulvis Talci etc.Solvent uses Purified Water, ethanol, propylene glycol etc. usually.Antiseptic generally adopts methaform, benzylalcohol, dihydroxy acetic acid etc.Antioxidant is more suitable with sulphite, ascorbic acid etc. usually.
Pericarp of Fructus Musae extract provided by the invention, its solution or powder can be added to be used in the goods oral.Pericarp of Fructus Musae extract also can combine with the Other Drinks raw material and make certain beverage product, as banana skin beverage, soda pop, fruit drink, lactobacillus beverage, sports drink, soyabean milk etc.Pericarp of Fructus Musae extract it is also conceivable that daily breads such as making cookies, chocolate, confection, chewing gum, snack, fruit jelly dessert, bread, bean curd, yoghourt.
Pericarp of Fructus Musae extract provided by the invention and contained active component thereof can be used as medicine, health food, food or food additive, are used to prevent and improve the depression that causes by loading, tired and hepatic injury and clinical symptoms thereof.Among the present invention, the dosage of pericarp of Fructus Musae extract can suitably be selected according to the conditions such as relative health status, age, sex and body weight of user.Adult's average weight is with 60 kilograms of calculating, and oral pericarp of Fructus Musae extract generally can be considered at 1 gram to 5 gram scopes in one day, part vic.There are not problems such as side effect more than certain 5 grams yet.
The present invention loads mice as the oxidative stress experimental model with Restraint Stress (restraint stress), estimate pericarp of Fructus Musae extract to (the oxygen radical absorbance capacity of ORAC in the mice plasma, ORAC) active (Chinese Pharmacological circular, 22:1015-1021,2006), malonaldehyde in the liver (MDA) content, the active influence of glutamate pyruvate transaminase in the blood plasma (ALT).The result shows that the anti-oxidative ability of mice that pericarp of Fructus Musae extract brings out loading is low, the blood plasma Glutamate pyruvate transaminase rises shows the significant effect of improving on the statistics.In the antioxidation in vitro experiment, pericarp of Fructus Musae extract also demonstrates stronger antioxidation.Based on The above results, confirm the establishment of the new purposes of pericarp of Fructus Musae extract of the present invention.
Can prove that according to above-mentioned experiment it is a kind of safe the invention provides pericarp of Fructus Musae extract, from the natural antioxidation of passing through, thereby reach anti-stress, antidepressant, alleviate and improve in response to swashing the untoward reaction such as hepatic injury that load causes, be used to prepare can take for a long time, medicine, food and the food additive of safe without toxic side effect.
In addition, the present invention also points out Pericarpium Musae to get thing and also can be used as antioxidant or antioxidation constituent raw material, is used to alleviate and improve the inflammatory reactions such as hepatic injury that cause by loading.Because pericarp of Fructus Musae extract provided by the invention is the material of natural origin, and among the peoplely have a long-term edible experience, be applicable to the crowd of each age level and health status.Therefore, pericarp of Fructus Musae extract can be used as health food, food additive and multiple use is provided.Have, although the Fructus Musae antidepressant effect is known, the antidepressant activity that Pericarpium Musae demonstrates confirms by the present invention is first again.
The specific embodiment:
The present invention can be elaborated by following examples, but is not limited only to following examples.
Embodiment one: the preparation of Pericarpium Musae water extract
Get fresh Pericarpium Musae, blend fully under 5 times of amount distilled water room temperatures of back adding and soaked 24 hours, gauze filters, and after centrifugalize in 3000 rev/mins, 15 minutes, reclaims supernatant.45 ℃ of concentrating under reduced pressure postlyophilizations promptly get the about 3.28% Pericarpium Musae water extract of the response rate and are used for following experiment embodiment of the present invention.
Embodiment two: the preparation of Pericarpium Musae alcohol hypostasis
After getting 30 times of amount distilled water of embodiment one Pericarpium Musae water extract adding, adding 95% ethanol to alcoholic acid final concentration is 80%, centrifugalize in 5000 rev/mins, 15 minutes, after getting the lyophilization of ethanol precipitation thing, the Pericarpium Musae alcohol hypostasis that promptly gets the response rate about 47.3% is used for following experiment embodiment of the present invention.
Embodiment three: the preparation of Pericarpium Musae alcohol soluble substance
After getting 30 times of amount distilled water of embodiment one Pericarpium Musae water extract adding, adding 95% ethanol to alcoholic acid final concentration is 80%, centrifugalize in 5000 rev/mins, 15 minutes, get the dissolve with ethanol supernatant, reclaim solvent behind the concentrating under reduced pressure, the Pericarpium Musae alcohol soluble substance that promptly gets the response rate about 41.2% is used for following experiment embodiment of the present invention.
Embodiment four: the Pericarpium Musae water extract is to the influence of Restraint Stress load mice plasma ORAC (ORAC) level
Restrain load model to be one of classical psychological stress model.This method stress (Neuropharmacology, 14:443-452,1996) by the purest setback of freely bringing out of depriving animal activity.Free activity by the constraint laboratory animal causes the lines of strain and fear psychology of animal.The depression model (Steriod, 65:8-15,2000) that often will restraining clinically to load is considered as stress-induced.This model can more effectively cause the intense strain of animal, also is irritability hepatic injury and the gastric ulcer model of using always (Acta Pharmaceutica Sinica, 41:631-635,2006).
Should swash the load experiment uses the male cleaning level of 7 18-22g in age in week Kunming mouse available from Guangdong Medical Lab Animal Center (quality certification SCXK (Guangdong) 2005A012).23 ± 2 ℃ of raising temperatures, lighting hours 12 hours/day (7:00～19:00 turns on light).After raising a week, experimentize.In the Restraint Stress experiment, according to 10ml/kg disposable gastric infusion before restraining.Press 500mg/kg, 250mg/kg and 125mg/kg Pericarpium Musae water extract aqueous solution oral administration, matched group is given and the equal-volume aqueous solution.7 every group of mices, every mice fasting forces to put into contained device, normal only fasting of blank group.Restrain after 12 hours under etherization, put into the centrifuge tube that heparin was handled after the heart blood sampling, through 5000 rev/mins of separated plasmas after centrifugal 5 minutes.Be after 3% perchloric acid removes albumen through final concentration again, reclaimed non-albumen supernatant in centrifugal 15 minutes by 15000 rev/mins and be used for ORAC and analyze.Blood plasma ORAC (ORAC) analysis is used multi-functional fluorescence analyser (Tecan Genios, Austria), excite with emission wavelength and be respectively 485nm and 525nm, the oxidant of attacking fluorescein (disodium fluorescein) uses 2,2 '-azobis-2-amidinopropane-dihydrochloride (AAPH; Wako Pure Chemical Industries, Ltd.Osaka, Japan).Potency unit is with reference to watermiscible vitamin E (6-hydro-2,5,7,8-tetramethylchroman-2-carboxylic acid; Trolox, Wako Pure Chemical Industries, Ltd.) (J Agric Food Chem., 51:3273-3279,2003).Experimental result is done statistical procedures with the t check.Experimental result is as shown in table 1, matched group is compared with loading, give blood plasma ORAC (ORAC) with the contained mice of Pericarpium Musae water extract, the ability that is the resistant activity oxygen-derived free radicals obviously strengthens, wherein 125mg/kg and 500mg/kg Pericarpium Musae water extract administration group are compared with the Stress model group and are had statistical significance (* p＜0.05), and have certain dose-effect relationship.
Table 1: the Pericarpium Musae water extract is to the influence of Restraint Stress load mice plasma ORAC level
* p＜0.05 has statistical significance with stress the load model matched group comparing.
Embodiment five: pericarp of Fructus Musae extract is to the influence of Restraint Stress load mice plasma and liver level of vitamin C
Laboratory animal, experimental technique of stress loading and experiment grouping are all identical with embodiment four with dosage.Mice is restrained and stress under etherization put into the centrifuge tube that heparin was handled by the heart blood sampling after 12 hours, through 5000 rev/mins of separated plasmas after centrifugal 5 minutes.Be that centrifugal 15 minutes of 12000rpm got supernatant and is used for ascorbic analysis after 10% metaphosphoric acid solution was handled through final concentration again.
Vitamin C working sample processing method in the hepatic tissue is, after getting the 0.1g tissue and adding 3% perchloric acid solution 1ml, is prepared into tissue homogenate in 30 seconds with IKAULTRA-TURRAXT8 type tissue refiner rotating speed homogenate with 15000 rev/mins under ice bath.Tissue homogenate and under 12000rpm centrifugal 15 minutes are got 20 times of supernatant dilutions and are used for HPLC and analyze.Vitamin C is measured and is used the HP1050 of Hewlett-Packard series of high efficiency chromatograph of liquid (comprising column oven, degasser, infusion pump, VWD UV-detector); Marco work station and C
18Chromatographic column (4.6 * 150mm; COSMOSIL Company).Mobile phase is 0.01mol/l potassium phosphate buffer (phosphoric acid is transferred pH value to 3): methanol (v: v=99: 1); Flow velocity is 1.0ml/min; The ultraviolet detection wavelength is 245nm.Experimental result is done statistical procedures with t check, and the vitamin C analysis result is as shown in table 2 in blood plasma and the liver: with stress compare by the load model matched group, obviously strengthen for the mice plasma level of vitamin C with the Pericarpium Musae water extract.250mg/kg and 500mg/kg Pericarpium Musae water extract administration group are compared with the Stress model group has statistical significance (* p＜0.05), and has certain dose-effect relationship.Give with the Pericarpium Musae water extract after stress mouse liver in vitamin C with stress compare by the load model matched group, also have outstanding property difference (* * p＜0.01) statistically, and have certain dose-effect relationship (table 3).
Table 2: the Pericarpium Musae water extract is to the influence of Restraint Stress load mice plasma level of vitamin C
* p＜0.05 has statistical significance with stress the load model matched group comparing.
Table 3: the Pericarpium Musae water extract is to the influence of Restraint Stress load mouse liver level of vitamin C
* p＜0.05 has statistical significance with stress the load model matched group comparing.
Embodiment six: the activity of the antioxidation in vitro Capability index (ORAC) of pericarp of Fructus Musae extract
The antioxidation in vitro experiment is that the pericarp of Fructus Musae extract that distinct methods prepares directly is added in the 96 hole ELISA Plate, and observation sample inhibited oxidation agent AAPH is to the delay action of the fluorescence decay of fluorescein (disodium fluorescein).As shown in Figure 1, the Pericarpium Musae water extract, Pericarpium Musae alcohol hypostasis and Pericarpium Musae alcohol soluble substance by the speed of reactive oxygen free radical cancellation, have the effect of resistant activity oxygen-derived free radicals at the external fluorescent material that all can delay.The resistant activity oxygen-derived free radicals effect that pericarp of Fructus Musae extract is described may come from and comprise the glycoprotein chemical compound that contains in the micromolecule polyphenol compound that contains in the Pericarpium Musae alcohol soluble substance and the Pericarpium Musae alcohol hypostasis.
Embodiment seven: the Pericarpium Musae water extract is to the mensuration of mda content in the liver
Body produces oxygen-derived free radicals by enzyme system and non-enzyme system can attack polyunsaturated fatty acid in the biomembrane, causes lipid peroxidation, and therefore forms lipid peroxidation product-malonaldehyde (MDA), causes cellular metabolism and dysfunction, even dead.Thereby the test MDA amount can reflect body inner lipid peroxidating degree, reflect the degree of cell damaged indirectly.
Mice behind the Restraint Stress load is taken out liver under the etherization state, produces 5% liver tissue homogenate with normal saline, 3000 rev/mins centrifugal 10 minutes, get the tissue homogenate supernatant and measure wherein mda content.Measuring principle utilize malonaldehyde to contract with thiobarbituricacid (TBA) and, form red product, in the character that there is maximum absorption band at the 532nm place, measure by colorimetry.Measure and use malonaldehyde to measure test kit (Nanjing is built up), be reflected in 96 orifice plates and carry out, use microplate reader (Finland's thunder is won the MK3 microplate reader) under 532nm, to measure absorptance (process of specifically measuring is referring to the test kit description).
Mda content in the computation organization needs the protein content in the known tissue.Therefore, produce 5% liver tissue homogenate with normal saline equally, utilize the protein content (use the protein quantification test kit, Nanjing is built up, and concrete operation method is seen the test kit description) in the Coomassie brilliant blue method mensuration tissue.Experimental result is as shown in table 4.Give with the Pericarpium Musae water extract and reduced in response to malonaldehyde (MDA) content in the liver tissue homogenate of swashing the load increase.With stress compare by the load model matched group, 500mg/kg Pericarpium Musae water extract administration group has significant difference statistically.
Table 4: the Pericarpium Musae water extract is to malonaldehyde in the liver (MDA) Determination on content
* p＜0.05: have statistical significance with stress the load model matched group comparing.
Embodiment eight: the Pericarpium Musae water extract is to the influence of blood plasma glutamate pyruvate transaminase (ALT) level
The vigor of blood plasma glutamate pyruvate transaminase is a characteristic index of hepatocyte injury degree.Stress can cause that a large amount of free radicals and cytokine produce.Therefore, these phenomenon explanations, at first the macrophage in the activating liver cell activates various cytokines then, causes inflammatory reaction, but the cytotoxicity of induction of lymphocyte, macrophage, by number of ways such as hepatocellular apoptosis damage hepatocyte.
The blood plasma of Restraint Stress mice is measured the vigor of glutamate pyruvate transaminase (ALT) wherein by reitman-frankel method.Measuring principle utilizes glutamate pyruvate transaminase (ALT) under 37 ℃ and PH 7.4 conditions, acts on the substrate that alanine and α-Tong Wuersuan are formed, and generates acetone acid and glutamic acid.After reaction a period of time, add 2,4 dinitrophenyl hydrazine (DNPH) hydrochloric acid solution, i.e. cessation reaction, carbonyl addition in DNPH and the keto acid simultaneously, generation acetone acid phenylhydrazone.Phenylhydrazone is rufous under alkali condition, colorimetry is measured.Measure in the concrete experiment and use malonaldehyde to measure test kit (Nanjing is built up), be reflected in 96 orifice plates and carry out, use microplate reader (Finland's thunder is won the MK3 microplate reader) under 532nm, to measure absorptance (process of specifically measuring is referring to the test kit description).This experimental result shows that compare and stress load model group plasma A LT level significantly raise with the normal control group, contained load brings out the experiment mice acute liver damage.Pericarp of Fructus Musae extract senior middle school low dose group all can be improved to a certain extent because of restraining the rising of the mice plasma ALT due to stress loading, wherein high dose and middle dosage group compare with the Stress model group show significant difference (
*P＜0.05).Small dose group also has the effect that reduces plasma A LT, but action effect does not have statistical significance.
Table 5: the Pericarpium Musae water extract is to the influence of blood plasma glutamate pyruvate transaminase (ALT) level
* p＜0.05: have statistical significance with stress the load model matched group comparing.
Embodiment nine: the Pericarpium Musae water extract is to the influence of mice forced swimming (forced swim test)
Forced swimming experiment (Eurp.J.Pharm, 47:379-391,1978) is based on when mice is compelled to put into a restricted waters, and the initial stage can be made great efforts swimming to flee from this environment.Take a kind of motionless passive state owing to escaping very fast just the drawing rein gradually of this environment, promptly only head above water is breathed.The fight behavior and because the desperate depressive emotion that causes that reflect mice by total dead time (total duration of immobility) of mice in the record experimentation.Therefore the desperate model of this behavior is widely used in the evaluation of depressed degree and the primary dcreening operation of antidepressant drug most antidepressants and central stimulants sensitivity.
In this experiment, we utilize mice forelimb climbing exercise and the good dependency of its swimming behavior, adopt specific forced swimming device (Model WW3002, OHC O ' hara ﹠amp; Co., Ltd., Tokyo, Japan).In the forced swimming experiment, the degree that the number of turns that we adopt the climbing action of computer recording mice forelimb to do the runner rotation that causes reflects the experiment mice depressive state can reflect the excitement or antidepressant sample effect (the Pharmacol Biochem Behav. of medicine more objectively than dead time of artificial observed and recorded mice, 56:131-137,1997), avoid the diversity of artificial judgement, more can reflect the effect of medicine objectively.Experiment divides to be carried out in three days, and first and second day is training Day.Being about to swim, water adds to pulley central authorities in the case, and about 18cm highly locates, and this water potential can not climb out of outside the case by pulley when guaranteeing mice swimming, also can be undue tired because of reaching pulley.Water temperature remains on 22～23 ℃.During training mice is put into water gently towards pulley, make its swimming take out to dry after 10 minutes and put back in the rearging cage, trained continuously two days.Weed out the mice of being overexcited or suppressing in the training process.Experiment day is divided into 4 groups with mice by body weight at random, 10 every group, is respectively distilled water blank group, 125,250 and 500mg/kg Pericarpium Musae water extract group.The Pericarpium Musae water extract all is dissolved in the distilled water gastric infusion by 10ml/kg before experiment.The swimming case is put into gently to pulley in 20 minutes back of mice administration, makes its swimming 6 minutes, the number of turns that adopts behind the computer recording 2 minutes mices to stir pulley with forelimb.Experiment, is carried out between 17:00 (afternoon) in 9:00 (morning) in 25 ℃ of rooms of room temperature in peace and quiet.
As shown in Figure 2, cause in the desperate behavioral experiment of mice at forced swimming, irritate stomach give with 125,250 and 500mg/kg Pericarpium Musae water extract all can increase the number of turns that the mice forelimb is stirred the runner rotation, and have certain dose-effect relationship.
Embodiment ten: the Pericarpium Musae water extract to the rat runner stress (activity stress) influence
The rat runner stress (activity stress) model (J Comp Physiol Psychol.47:362-363,1954) be rat to be put into runner make its autonomic movement.In this model, stress load applies by dietary restriction and runner two aspects of moving.Restrictive diet combines with the runner motion and makes rat engender that the runner quantity of motion increases, interaction between this restrictive diet and the runner motion is believed to cause animal to enter dying state (Brain research by increasing activity, 891:32-41,2001).This characteristics that stress load are that the runner quantity of motion that animal self improves constantly has constantly applied bigger stress stimulation to self.This model can bring out digestive tract ulcer and multiple gastrointestinal disease (J Pharmacol Exp Ther.273:1001-1007,1995), but its main analog neuron apositia (Int.J.Eat.Disord.3:27-46 that is applied in; 1983).Neural biochemical indicator has also further proved the similarity (Physiol Behav.52:299-304,1992) between this model and neural apositia and the obsession.
Male cleaning grade SD rat is used in the experiment of stress loading, and (body weight 220～260g) provides (quality certification number: SCXK (Guangdong) 2003-2002) by Guangdong Medical Lab Animal Center.Animal feeding condition: 23 ± 2 ℃ of raising temperatures, lighting hours 12h/d (7:00～19:00 turns on light).Carry out every experiment after raising a week.With experimental rat by body weight divide at random normal control group, Stress model matched group and 250 and 500mg/kg/day Pericarpium Musae water extract-runner stress the load group.Every group 7 the animal 18:00-20:00 every day gastric infusions and the drinking-water of taking food continue 14 days record rat motor states.The runner stress rats taken out in the 15th day, etherization, postcava are got behind the blood immediately broken end and are got different brains district tissue sample, measure the monoamine neurotransmitter level in the tissue.
Rat carries out after rotating cage stress load, quantity of motion continues to increase along with the increase of the natural law of stress loading, and increase in wherein preceding 4 days is slower, and the trend that appearance is increased fast after 4 days, a part of laboratory animal begins to occur phenomenon in the flue after 14 days, progressively is on the verge of death.The Pericarpium Musae water extract and stress be compared by matched group to the influence of runner exercise stress rat motor amount as shown in Figure 3,250 and 500mg/kg Pericarpium Musae water extract increasing of runner number had certain mitigation, and have certain dose-effect relationship.
Embodiment 11: the Pericarpium Musae water extract is to the influence of 5-hydroxy tryptamine (5-HT) level in the runner load rat brain
Runner stress be loaded after the experiment end, experimental rat breaks end after abdominal vein is got blood under the etherization condition at once, take out full brain, and on ice pan, separate cerebral cortex, Hippocampus and hypothalamus rapidly, remove blood with pre-cold saline rinsing, weigh after blotting on the filter paper, place the 1.5ml round bottom centrifuge tube.Rat cerebral cortex, Hippocampus and hypothalamus added 0.1mol/L perchloric acid in molten 1: 1,1: 2 and 1: 5 by anharmonic ratio, tissue homogenate is made in tissue refiner in ice bath, centrifugal 20 minutes of 4 ℃ of 15000rpm, supernatant-80 ℃ of freezing preservations of refrigerator behind 0.2 μ m membrane filtration are used to detect the 5-HT level.
High performance liquid chromatogram coulomb array electrochemical detection system is a U.S. ESA company, comprises Model 582 infusion pump, Model 542 automatic samplers, Model 5600A CoulArrayDetector coulomb array electrochemical detector and The CoulArray for windows-based workstation software.Chromatographic column adopting Develosil ODS-UG-5 (4.6mm * 150mm, NOMURA CHEMICAL), guard column are the (4.0Lmm * 3.0LDmm) of U.S. Phenomenex company.Mobile phase is formulated as title-hydration citric acid 15.0g, trisodium citrate 20.2g, and EDTANa2 0.036g, eight sodium alkyl sulfonate 0.18g add chromatograph pure water (〉=18M Ω) 900ml; Add 100ml methanol, ultrasonic degas is standby behind the 0.2 μ m membrane filtration.Column temperature is set at 38 ℃, and flow velocity is adjusted into 0.8ml/min.Coulomb array electrochemical detector is set three channel electrode electromotive forces: passage 1:-150mV (reduction potential); Passage 2:+350mV (NE, DOPAC, DA, 5-HIAA, 5-HT sense channel); Passage 3:+450mV (HVA sense channel).
Stress the load model group compare with runner, the Pericarpium Musae water extract is not obvious for 5-HT content effect in the cerebral cortex.But, 250 and the 500mg/kg banana oil peel extract 5-HT content (P＜0.05) in hypothalamus and the Hippocampus that can raise.
Table 6: the Pericarpium Musae water extract is to the influence of 5-hydroxy tryptamine (5-HT) level (ng/g)
* p＜0.05: have statistical significance with stress the load model matched group comparing.
5-HT participates in emotional response, and Barnes people's such as (Pharmacol Biochem Be, 58:775-783,1997) research confirms that also various acute stresses stimulate the release that can promote 5-HT in the brain.Our experiment shows that stress stimulation and pericarp of Fructus Musae extract are promoting that 5-HT is synthetic, having cooperative effect on release and the metabolism.
The rat runner stress be analog neuron apositia, compulsive animal model (Physiol Behav.51:827-832,1992).Nervous anorexia is a kind of and nervus centralis, the relevant abnormal food appetite disease of the mental status, and is relevant with psychological factor.(obsessive-compulsivedisorder OCD) is meant that to force symptom be the neurosis of main clinical manifestation to obsession.Force symptom be consciousness on the HA symptom.In recent years studies have shown that obsession is relevant with the 5-HT function.Our experimental result shows that the Pericarpium Musae water extract increases 5-HT content in stress rats hypothalamus, the Hippocampus, and prompting Pericarpium Musae water extract may be relevant with adjustment 5-HT effect for the ethological improvement effect of runner stress rats.
The present invention stress load mice as the oxidative stress test model to restrain, and estimates the anti-stress effect of pericarp of Fructus Musae extract.The result shows that the pericarp of Fructus Musae extract of various dose can effectively reduce the MDA content of liver tissue homogenate, reduces plasma A LT activity.Therefore infer that the anti-stress of pericarp of Fructus Musae extract, alleviation and improvement prevent peroxide injury by untoward reaction effect and the antioxidation such as hepatic injury that stress load and cause, the effects such as generation that reduce lipid peroxide are relevant.In addition, the present invention also points out pericarp of Fructus Musae extract also to can be used as antioxidant or antioxidation constituent raw material, is used for some because the prevention and the auxiliary treatment of the disease that oxidation causes.Based on The above results, confirm the establishment of the new purposes of pericarp of Fructus Musae extract of the present invention.
Can prove that according to above-mentioned experiment it is a kind of safe the invention provides with pericarp of Fructus Musae extract, from the natural antioxidation that passes through, thereby reach anti-stress, antidepressant, alleviation and prevention, improvement and treatment are used to prepare medicine that can take for a long time, safe without toxic side effect, food additive, health food and constituent thereof by the untoward reaction such as hepatic injury that stress cause.In addition, the present invention also points out pericarp of Fructus Musae extract also to can be used as antioxidant or antioxidation constituent raw material, is used to alleviate and prevent The symptoms of aging such as skin aging, senile cataract and alzheimer disease.Owing to the invention provides the material that pericarp of Fructus Musae extract is a natural origin, and among the people secular edible experience arranged, be not only applicable to general adult, also be applicable to the low person of old people, child and muscle power.Therefore, with pericarp of Fructus Musae extract can be used as health food, food additive provides multiple use.Can prove that according to above-mentioned experiment Pericarpium Musae water extract provided by the invention is a kind of safe, from the natural antioxidation that passes through, thereby reach anti-stress, alleviate and improve, be used to prepare medicine that can take for a long time, safe without toxic side effect, food and food additive by the untoward reaction such as hepatic injury that stress cause.
Description of drawings
Fig. 1 is antioxidation in vitro Capability index (ORAC) the measurement result analysis chart of pericarp of Fructus Musae extract;
Fig. 2 is that pericarp of Fructus Musae extract is to the momental figure of influence of mice forced swimming;
Fig. 3 be the Pericarpium Musae water extract to the rat runner stress influence figure.
Claims (7)
1. be that effective ingredient is used for anti-stress with the pericarp of Fructus Musae extract, antidepressant, the purposes of the medicine of the untoward reaction such as hepatic injury that alleviation and improvement cause by loading.
2. be that effective ingredient is used to prepare anti-stress with the pericarp of Fructus Musae extract, the health food of the untoward reaction such as hepatic injury that cause by loading and the purposes of food additive are alleviated and improved to antidepressant.
3. purposes according to claim 1 and 2 is characterized in that: described pericarp of Fructus Musae extract contains polysaccharide and polyphenol active ingredient, many 5 milligrams of the whats of the content of polysaccharide and polyphenol active ingredient in every gram pericarp of Fructus Musae extract.
4. purposes according to claim 1 and 2 is characterized in that: described pericarp of Fructus Musae extract is to adopt one or more any mixed solvent of water or water and lower alcohols, polar solvent to extract.
5. purposes according to claim 1 is characterized in that: described medicine comprises the pericarp of Fructus Musae extract of dose therapeutically effective, and pharmaceutically acceptable carrier.
6. purposes according to claim 1 and 2 is characterized in that: described pericarp of Fructus Musae extract dry product use amount in the various preparation finished products of medicine or health food or food additive can account for 0.001～100% weight ratio.
7. purposes according to claim 4 is characterized in that: described pericarp of Fructus Musae extract can add sodium bicarbonate and/or SODIUM ASCORBATE in solvent when extracting.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105639400A (en) * | 2015-12-30 | 2016-06-08 | 湖北文理学院 | Banana peel antioxidant additive and preparation method thereof |
| CN108578531A (en) * | 2018-03-26 | 2018-09-28 | 广西玉阳绿州生物科技有限公司 | Intestinal cancer healing liquid and its production technology |
| CN110113957A (en) * | 2016-12-01 | 2019-08-09 | 财团法人全南生物产业振兴院 | The health functional food composition for being used to prevent and improve depression comprising blueberry fruit extract |
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2007
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105639400A (en) * | 2015-12-30 | 2016-06-08 | 湖北文理学院 | Banana peel antioxidant additive and preparation method thereof |
| CN110113957A (en) * | 2016-12-01 | 2019-08-09 | 财团法人全南生物产业振兴院 | The health functional food composition for being used to prevent and improve depression comprising blueberry fruit extract |
| CN108578531A (en) * | 2018-03-26 | 2018-09-28 | 广西玉阳绿州生物科技有限公司 | Intestinal cancer healing liquid and its production technology |
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