KR20190085565A - Composition for fatigue recovery comprising Pinus koraiensis leaves extract as effective component - Google Patents

Abstract

The present invention relates to a composition for fatigue recovery comprising a Pinus Koraiensis leaf extract as an effective component. When a Pinus Koraiensis leaf extract which is an effective component is injected into an animal model with fatigue and stress induced by forced swimming, the content of lactate, BUN, NH_3 and LDH in serum increased by excessive exercise (forced swimming) is reduced and the content of cortisol, an elevated stress indicator due to excessive exercise, is reduced. Accordingly, the Pinus Koraiensis leaf extract can be usefully used as a health functional food composition for fatigue recovery.

Description

[Technical Field] The present invention relates to a composition for recovering fatigue comprising an extract of Pinus koraiensis as an active ingredient,

The present invention relates to a composition for restoring fatigue comprising an extract of pine tree leaf as an active ingredient.

Fatigue is generally defined as a state of abnormal burning after routine activities, a state in which there is no aura and can not perform a continuous effort or concentration, or a general lack of energy to perform routine activities. Is called prolonged fatigue if it lasts more than one month, and chronic fatigue if it lasts longer than six months.

Chronic Fatigue Syndrome, unlike transient fatigue that recovers from a short break, continues to cause fatigue, which makes the patient very debilitated without taking a rest. The cause of chronic fatigue syndrome has yet to be clarified. However, related diseases include various infectious diseases including virus infection, transient trauma or shock, extreme stress, and toxic substances. Recently, it has been argued that the disorder is caused by central nervous system disorder. The reason is that the symptoms such as concentration disorder, attention disorder, memory disorder, and sensory abnormality occur frequently in patients with chronic fatigue syndrome, % Of patients present with symptoms such as temporary paralysis, visual impairment, mismatching, or confusion within the first 6 months after the onset of the disease. Since these symptoms occur when an abnormality occurs in the central nervous system, it suggests a link between the chronic fatigue syndrome and the central nervous system. In addition, patients with chronic fatigue syndrome who have decreased cerebral blood flow is found in the SPECT test, or abnormal findings of various neurotransmitters have been found that the theory has been raised, but the cause is not yet clear.

On the other hand, pine wood is considered as a major forest resource, but it is not utilized separately except for the nut of pine and wood. Especially, the shingled leaf has not only a wax layer on the surface of the leaves but also a strong taste. The shingles are called "柏葉" in the oriental medicine. It contains 0.25 ~ 0.41% essential oil, 350mg% flavonoid, 260 ~ 360mg% ascorbic acid, which is known to strengthen the digestive system. In the case of folk remedies, fresh shrimp leaves were washed with water, then water was removed, the leaves were poured in water, and a lobster tea boiled in water was used to treat diarrhea and dysentery in children. In order to improve the palatability of the lobster tea used as the folk remedy, the liquid obtained by dissolving the sham-leaf and the sugar in the jar was aged in a cool place and diluted with water was used.

Prior art related to the pine leaf extract has been disclosed in Korean Patent Publication No. 2012-0012639, a beverage containing a pine tree extract and a preparation method thereof, and Korean Patent Publication No. 2011-0056053 discloses separation And an extract thereof are disclosed, but no fatigue recovery composition containing the extract of Pine Pinaceae of the present invention as an active ingredient has been disclosed.

The present invention is derived by the request as described above, the present invention is pine (Pinus The present invention provides a composition for restoring fatigue comprising leaf extract as an active ingredient and is useful as an active ingredient for the stress and fatigue induced ICR mice, (NH ₃ ), blood urea nitrogen (BUN), lactate dehydrogenase (LDH), and cortisol content in the blood were decreased. As a result, the present invention .

In order to achieve the above object, the present invention provides a health functional food composition for restoring fatigue comprising an extract of P. vivax as an active ingredient.

The present invention also provides a method of restoring fatigue comprising the step of administering the composition to a non-human subject.

The present invention relates to a composition for restoring fatigue comprising an extract of Pine Pinus densiflora as an active ingredient, which comprises lactate, ammonia (NH ₃ ), blood urea nitrogen (BUN), lactate dehydrogenase (LDH) and cortisol , As well as the effect of reducing stress and fatigue activity in forced swimming animal models.

Fig. 1 shows that 100 mg / kg (PKE-100), pine tree leaf extract 200 mg / kg (PKE-200) and 100 mg / kg for 10 days, and the body weight was measured on days 1, 5, and 10 (#: p <0.05 vs. vehicle).
Fig. 2 shows the results of a 10-day administration of 100 mg / kg of pine tree leaf extract (100 mg / kg) (PKE-100) As a result of the test, it shows the swimming duration in water (##: p <0.01, ###: p <0.001 vs. vehicle).
FIG. 3 is a graph showing the results of stress and fatigue induced ICR mice induced by forced swimming in 100 mg / kg (PKE-100), 200 mg / kg (PKE-200) kg for 10 days and the concentration of lactate in the blood obtained on day 11 was measured by lactic acid oxidase electrode method (#: p <0.05, ##: p <0.01, #### : p < 0.0001 vs. vehicle).
FIG. 4 shows the results of stress and fatigue-induced ICR mice induced by forced swimming in 100 mg / kg (PKE-100), 100 mg / kg (PKE-200) kg for 10 days, and the concentration of ammonia (Ammonia) in the blood obtained on the 11th day was colorimetrically measured (####: p <0.0001 vs. vehicle).
FIG. 5 shows the results of stress and fatigue induced ICR mice induced by swimming with 100 mg / kg (PKE-100), pinewood leaf extract 200 mg / kg (PKE-200) kg) for 10 days and the concentration of blood urea nitrogen (BUN) in the blood obtained on the 11th day was colorimetrically measured (#: p <0.05 vs. vehicle).
FIG. 6 shows the results of stress and fatigue induced ICR mice induced by forced swimming in 100 mg / kg (PKE-100), 100 mg / kg (PKE-200) kg for 10 days, and the concentration of lactate dehydrogenase (LDH) in the blood obtained on day 11 was measured by colorimetry (###: p <0.001 vs. vehicle).
Figure 7 shows the results of stress and fatigue-induced ICR mice induced by forced swimming in 100 mg / kg (PKE-100), pinewood leaf extract 200 mg / kg (PKE-200) kg for 10 days, and the concentration of cortisol in the blood obtained on the 11th day was confirmed by enzyme immunoassay (#: p <0.05 vs. vehicle).

The present invention relates to a health functional food composition for restoring fatigue comprising a pine cone leaf extract as an active ingredient.

The Pine leaf blade extract may be prepared by a method including, but not limited to, the following steps:

(1) extracting a dry powder of pine tree by adding an extraction solvent;

(2) filtering the extract of step (1); And

(3) The step of extracting the filtered extract of step (2) by concentration under reduced pressure and drying.

In the step (1), the extraction solvent is preferably selected from water, a C ₁ -C ₄ lower alcohol or a mixture thereof, more preferably ethanol, even more preferably 80% (v / v) ethanol But is not limited thereto.

In the above production method, any conventional method known in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction may be used. The extraction solvent is preferably added by 1 to 20 times the weight of the dried pine tree leaves, more preferably 5 to 15 times. The extraction temperature is preferably 4 to 100 DEG C, but is not limited thereto. The extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 6 hours, most preferably 4 hours, but not always limited thereto. In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in the step (3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

The fatigue is characterized by being fatigue caused by muscle fatigue or stress caused by exercise, and muscle fatigue may be caused by fatigue, decreased endurance, impaired ability to work, or lethargy.

The health functional food composition for fatigue recovery according to an embodiment of the present invention may further include at least one selected from the group consisting of taurine, creatine, vitamin E, vitamin C, carotenoid, reduced glutathione, and minerals in addition to the active ingredient.

The composition is preferably prepared by any one of powder, granule, ring, tablet, capsule, candy, syrup and beverage, but is not limited thereto.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is, or may be used together with other food or food ingredients, and suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably used depending on the purpose of use (prevention or improvement). Generally, the health functional food composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight based on the raw material, when the food or beverage is produced. However, in the case of long-term intake intended for health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount of more than the above range.

There is no particular limitation on the kind of the health functional food. Examples of foods to which the health functional food composition can be added include meat products, sausages, breads, chocolate, candies, snacks, confectionery, pizza, ramen noodles, other noodles, gums, dairy products including ice cream, various soups, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

In addition, the health functional food composition of the present invention can be produced as a food, particularly a functional food. The functional food of the present invention includes components that are ordinarily added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when it is made of a drink, it may contain, in addition to the active ingredient, a natural carbohydrate or a flavoring agent as an additional ingredient. The natural carbohydrate may be selected from the group consisting of monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g., dextrin, cyclodextrin, For example, xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above health functional food composition, it is also possible to use various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in beverages, and the like. Although the ratio of the above-mentioned ingredients is not critical, it is generally selected in the range of 0.01 to 0.1 part by weight based on 100 parts by weight of the health functional food composition of the present invention.

The invention also relates to a method of restoring fatigue comprising the step of administering a composition according to the invention to a non-human subject.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

1. Experimental material

The honggyeongcheon extract (Rhodiola used as a positive control (PC) rosea extract was purchased from SK Bioland, and CMC (carboxymethylcellulose sodium) was purchased from Sigma (St. Louis, Mo. USA).

2. Preparation of extract

Pinus pinus koraiensis) into a 80% (v / v) ethanol, corresponding to 10 times the leaf weight, and extracted under reflux at 70 ℃ for 4 hours. After extraction, the mixture was filtered using a filter paper, concentrated under reduced pressure using a distillation apparatus, lyophilized, and then chlorophyll removed to prepare a pine needle leaf extract (PKE).

3. Fatigue induced by stress-induced weight-loaded forced swim

Male ICR mice at 5 weeks of age were supplied from Korea BiLink (Korea). Animals were fed with solid feed (no antibiotics, Samyang Feed Co.) and water until the day of experiment, 55 ± 15%, 12 hour light-dark cycle, and adapted for 1 week. In order to induce stress and fatigue, 5% of body weight was attached to the tail of a mouse, and a total of 4 times of weight load forced swimming was performed once a day for 30 minutes before the test day.

4. Administration of the extract

All of the extracts were prepared by using physiological saline containing 0.5% CMC as a vehicle, and stored at -20 ° C. until administration, and were dissolved just before administration.

Oral administration was carried out 10 days before the start of the exercise. The control group was administered 0.1 ml of physiological saline containing 0.5% CMC, and the pine needle leaf extract administration group (PKE-100, PKE-200) administered 100 mg / kg and 200 mg / kg pine needle leaf extract Respectively. The positive control (PC) was administered with 0.1 ㎖ of 100 mg / kg Rhodiola rosea extract.

5. Weight measurement

The body weights of ICR mice were measured at 1, 5, and 10 days at constant time, and measurements were made with precision scales in the experimental animal room.

6. Weight-Loaded Forced Swimming Test

The water of the ICR mice was immersed in water to a depth of 25 ± 1 ° C so that the tail of the ICR mice did not touch the bottom of the cylindrical water tank (Visionscientific Co., Korea) made of transparent acrylic for the forced swimming test. The duration of swimming was 10 minutes after the end of the administration, 1 hour after attaching weight to the ICR mouse tail by 5% of the weight and using video tracking software (SMART 3.0, Panlab, Spain) And the time of sinking for 5 seconds or longer was measured and evaluated.

7. Analysis of biochemical components in plasma

Blood samples for biochemical analysis were collected by forced swimming without weight load for 30 minutes on the 11th day of the experiment. Lactate was measured by lactic acid oxidase electrode method using Lactate Pro2 (Arkray, Japan) as the collected blood. Ammonia, blood urea nitrogen (BUN) and lactate dehydrogenase (LDH) were collected by centrifugation at 1,000 × g for 10 min in an anticoagulant tube containing heparine Plasma was isolated and measured by colorimetry using a blood analyzer (Fuji Dri-Chem 7000i, Japan).

8. Plasma Cortisol ( Cortisol ) Content measurement

Cortisol content was measured in plasma separated from experimental animals. Cortisol was measured by a competitive enzyme immunoassay using the Cortisol ELISA Kit (Cayman Chemical Company, Ann Arbor, MI, USA). Cortisol AChE tracer and ELISA monoclonal antibodies were added to 50 μl of standard material and sample (plasma). Thereafter, the reaction was carried out overnight at 4 ° C., followed by washing. Ellman's reagent was added thereto for 2 hours, and the absorbance at 405 nm was measured.

9. Statistical processing

All data were expressed as mean ± SD (mean ± SD). The results of each experimental group were statistically analyzed using one-way ANOVA (Tukey's multiple comparison test), and a significance test was performed at a level of p <0.05 or less (#: p <0.05, ##: p < 0.01, ###: p <0.001, ####: p <0.0001 vs. vehicle).

Example  1. Stress and fatigue improvement effect of Pine leaf extract

In Example 1, the stress and fatigue-improving effect of the extract of P. fomulus leaf was confirmed by the above-mentioned experimental method. In detail, according to the above experimental method, physiological saline containing 5% CMC was administered to the ICR mice for 10 days to induce stress and fatigue. In the PKE group, 100 mg / kg (PKE-100) and 200 mg / kg (PKE-200). The positive control (PC) was administered with 100 mg / kg of Hongkryung Stream extract. One hour later, a weight corresponding to 5% of the mice 'weight was attached to the tail of the mice, and the animals were subjected to forced swimming for a total of 4 times for 30 minutes every day until the test day to induce stress and fatigue. One hour after the administration on the 10th day, the weight of 5% of ICR mice was added to the tail of the ICR, and the swimming duration was measured by the weight load forced swimming test. The weight change was observed after the experiment, Related physiological target substances were examined.

1) Confirm weight change

Effects of pine needle leaf extract on body weight of stressed and fatigued ICR mice.

As a result, as shown in Fig. 1, the body weight decreased on the 5th day as a whole. On the 10th day, the body weight of the pine needle leaf extract administration group (PKE-100, PKE-200) And it was confirmed that the body weight was restored.

2) Effect on swimming duration

Weight load forced swimming test is a widely used test to confirm fatigue improvement effect. When you go into the water during swimming, the body temperature is lowered and the heat production is increased, so a lot of heat is consumed, and most of the muscle movements are made in the skeletal muscle, which causes stress and fatigue and decreases the exercise capacity.

To investigate the effects of pine tree leaf extract on stress and fatigue induced swimming time in ICR mice, weight load forced swimming test was performed.

As a result, as shown in FIG. 2, the swimming duration of 100 mg / kg of pine needle leaf extract-treated group and the positive control group (PC) was significantly increased compared to the negative control group.

3) Effect on lactic acid content

If the oxygen supply during the high intensity exercise is less than the oxygen consumption of the muscles, the lactic acid concentration in the muscle tissue is increased, and the lactic acid produced is diffused into the blood to be treated in the heart and liver, resulting in internal body acidification and fatigue .

Therefore, the effect of the extract of P. intestinalis, an active ingredient of the present invention, on the content of lactate accumulated in the blood of stressed and fatigued ICR mice was examined. Specifically, the blood of ICR mice induced by stress and fatigue was measured by lactic acid oxidase electrode method using Lactate Pro2 (Arkray, Japan) according to the above experimental method.

As a result, as shown in FIG. 3, the amount of lactic acid was significantly decreased in the group treated with the extract of Pinus koraiensis (PKE-100, PKE-200) compared to the negative control group.

4) Effect on Ammonia Content

Aspartic acid is a major component of the purine nucleotide circuit in the muscle, regenerating the ATP pool during intense high-intensity exercise, providing energy to the muscle and forming free ammonia in the process. Depending on the strength or time of exercise, the ammonia produced as a result of the degradation of amino acids is released from the muscle into the blood, and the blood ammonia is transferred to the brain to increase the central fatigue. On the other hand, the accumulation of ammonia in muscle cells stimulates the afferent nerves that are associated with muscle pain sensation, inhibits the TCA cycle and glycogenesis, and causes muscle fatigue by causing lactic acid production.

Thus, the effect of the extract of P. vannamei on the ammonia content in blood of ICR mice induced by fatigue was examined. Specifically, the blood of ICR mice induced by stress and fatigue was recovered and centrifuged according to the above-mentioned experimental method. The serum portion was measured by a colorimetry method using a blood analyzer (Fuji Dri-Chem 7000i, Japan) to measure the ammonia content Respectively.

As a result, as shown in Fig. 4, the ammonia concentration was significantly decreased in the pine needle leaf extract administration group (PKE-100, PKE-200) as compared with the negative control group (Vehicle).

5) Blood element nitrogen  Effect on content

Urea is the final product of protein metabolism in the body. After intense exercise, protein and amino acid catabolism occurs and urea nitrogen is increased in the state of insufficient supply of energy by saccharide or lipidation. Therefore, it can be seen that the increase of blood urea nitrogen induced the catabolism and fatigue of protein or amino acid.

The effects of Pine needle leaf extract on the blood urea nitrogen (BUN) content in the blood of stressed and fatigued ICR mice were examined. Specifically, the blood of ICR mice induced by stress and fatigue was collected and centrifuged, and the serum portion was analyzed by colorimetry using a blood analyzer (Fuji Dri-Chem 7000i, Japan) to determine the blood urea nitrogen content Were measured.

As a result, as shown in FIG. 5, the concentration of blood urea nitrogen (BUN) in the pine needle leaf extract administration group (PKE-100, PKE-200) was significantly lower than that of the negative control group.

6) Lactate dehydrogenase  Effect on content

Lactate dehydrogenase plays a role in regulating the production and conversion of lactic acid in muscle cells during muscle activity, increasing post - exercise activity, and increasing the activity of lactate dehydrogenase in blood is an indicator of cell damage.

Effects of Pinus koraiensis L. extract on the lactate dehydrogenase (LDH) content in the blood of stressed and fatigued ICR mice. Specifically, the blood of ICR mice induced by stress and fatigue was collected and centrifuged, and the serum portion was analyzed by colorimetry using a blood analyzer (Fuji Dri-Chem 7000i, Japan) And the content thereof was measured.

As shown in FIG. 6, the concentration of lactate dehydrogenase (LDH) was significantly decreased in the pine needle leaf extract-treated group (PKE-100, PKE-200) compared to the negative control group.

7) Cortisol  Effect on content

Cortisol is a steroid hormone secreted from the adrenal cortex and is involved in the regulation of the response and homeostasis of individuals to stressors. Cortisol increases micturition stress and increases blood pressure and pulse rate. The higher the concentration of cortisol, the higher the stress.

The effect of Pine needle leaf extract on the Cortisol content in the blood of stressed and fatigued ICR mice was examined. Specifically, the blood of the ICR mice induced by stress and fatigue was collected and centrifuged, and the serum portion was analyzed by enzyme immunoassay using a Cortisol ELISA Kit (Cayman Chemical Company, Ann Arbor, MI, USA) And the content of cortisol was measured.

As a result, as shown in FIG. 7, the concentration of cortisol was significantly decreased in the pine needle leaf extract-administered group (PKE-100, PKE-200) compared to the negative control group.

Claims (6)

A health functional food composition for restoring fatigue comprising an extract of P. vivax as an active ingredient. The health functional food composition for restoring fatigue according to claim 1, wherein the solvent of the extract of P. perilla is water, a C ₁ -C ₄ lower alcohol or a mixture thereof. The health functional food composition for restoring fatigue according to claim 1, wherein the fatigue is fatigue caused by muscle fatigue or stress caused by exercise. The health functional food composition for fatigue recovery according to claim 1, further comprising at least one selected from the group consisting of taurine, creatine, vitamin E, vitamin C, carotenoid, reduced glutathione and minerals in addition to the active ingredient. The health functional food composition for restoring fatigue according to claim 1, wherein the composition is prepared from any one of powder, granule, ring, tablet, capsule, candy, syrup and beverage. A method of restoring fatigue comprising administering a composition according to any one of claims 1 to 5 to a non-human subject.

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