JP2006137715A - Vascular endothelium-derived hyperpolarizing factor enhancer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a means for safely and efficiently relaxing resistance vessels such as arterioles by enhancing a vascular endothelium-derived hyperpolarizing factor and to provide a novel method for treating or preventing circulatory disorders. <P>SOLUTION: This invention provides a vascular endothelium-derived hyperpolarizing factor enhancer containing an extract of leaves of hardy rubber tree (Eucommia ulmoides Oliver). This invention further provides a medicinal composition, a food composition, a food, a drink, or the like containing the vascular endothelium-derived hyperpolarizing factor enhancer. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present application relates to a vascular endothelium-derived hyperpolarizing factor enhancer comprising a nakanaka leaf water extract, and more particularly to a vascular endothelium-derived hyperpolarizing factor enhancer having a relaxing action in resistance vessels. Furthermore, the present invention relates to a composition for oral intake, a pharmaceutical composition, a food composition and the like containing a vascular endothelium-derived hyperpolarizing factor enhancer.

  Vascular endothelial cells not only act as a barrier for substance exchange inside the blood vessels, but also respond to physical stimuli caused by blood flow and neurotransmitters such as acetylcholine (hereinafter also referred to as “ACh”). It plays an active role in effectively supplying blood to organs by synthesizing and releasing vasoactive substances (Non-patent Document 1: Experimental Medicine, Vol. 18, No. 5, 2000 ( (Special Publication), 595-602; Non-Patent Document 2: Circulation Control, Vol. 22, No. 3, 2001, 186-193). As endothelium-derived vasodilators, nitric oxide (hereinafter also referred to as “NO”) produced by nitric oxide synthase (hereinafter also referred to as “eNOS”) and prostacyclin produced by cyclooxygenase were identified. ing. On the other hand, it has been observed that hyperpolarization of smooth muscle cells by muscarinic receptor stimulation induces vasorelaxation in various blood vessels, but it has been reported that it is not affected by eNOS inhibitors and cyclooxygenase inhibitors. Yes. Therefore, it is considered that hyperpolarization of smooth muscle cells occurs via an endothelium-derived hyperpolarizing factor (hereinafter also referred to as “EDHF”), which is different from NO and prostacyclin (non-patent literature). 2).

  Although EDHF has not yet been specifically identified, it is understood that EDHF plays a physiologically important role as a factor that dynamically regulates vascular tone. The endothelium-dependent relaxation reaction in the membranous artery, femoral artery, renal artery, etc. is mainly relaxation via EDHF (Non-patent Document 2). On the other hand, the endothelium-dependent vasorelaxation reaction via nitric oxide is peculiar to blood vessels with large diameters such as the aorta. Therefore, the vascular relaxation effect via EDHF in the endothelium-dependent vasorelaxation reaction can be expected to be a relaxation action specific to resistance blood vessels such as arterioles, and is effective for the treatment or prevention of microcirculation disorders and coldness. It can be a vasorelaxant effect. In addition, as vasodilators for medical use, ACE inhibitors, Ca antagonists, antiplatelet drugs, coronary artery dilators such as nitrate drugs, α-blockers, etc. are used, but each may have unique side effects. There is a need for vasodilators that are known and have a different mechanism of action.

Naturally derived foods and traditional Chinese medicines have advantages such as generally having few side effects, so their usefulness for various intractable diseases whose occurrence has been increasing in recent years has been attracting attention. Among them, tanaka bark has long been used as a Chinese herbal medicine in China and has been known to have central inhibitory action, antihypertensive action, diuretic action, and tanaka bark extract has an endothelium-dependent vascular relaxation effect. (Non-Patent Document 3: Naunyn-Schmiedeberg's Arch Pharmacol, 369, 2004, pp. 206-211). In addition, it has been reported that the Tochu leaf extract also has an endothelium-dependent vasorelaxing effect, but only a pathway through nitric oxide has been confirmed (Non-patent Document 4: Vascular Pharmacology, No. 40). Vol., 2004, pp. 229-235), there was no report on the relaxation reaction of the resistance blood vessels via EDHF. In particular, there has been no report on the endothelium-dependent vascular relaxation effect of the water extract of Tochu tea leaves obtained by processing Tochu leaves.
Experimental Medicine, Vol. 18, No. 5, 2000 (extra number), 595-602 Circulation control, Vol. 22, No. 3, 2001, pp. 186-193 Naunyn-Schmiedeberg's Arch Pharmacol, 369, 2004, 206-211. Vascular Pharmacology, 40, 2004, pp. 229-235

  As a result of diligent research to solve the above-mentioned problems, the present inventor discovered that the water extract of Tochu tea leaves has a specific enhancing action on vascular endothelial hyperpolarizing factor. Completed.

An object of the present invention is to provide a vascular endothelium-derived hyperpolarizing factor enhancer, a composition for oral ingestion, a pharmaceutical composition, a food composition, a food and a beverage containing a licorice leaf water extract.
That is, according to one aspect of the present invention, there is provided a vascular endothelium-derived hyperpolarizing factor potentiator comprising a foliage extract. Here, the bamboo leaf used in the present invention may be produced by cultivation or collected from nature. For example, fresh leaves before the fall of the current year are used, and fresh leaves from April to October, preferably from May to August, more preferably from June to August can be used. The nakanaka leaf used for water extraction may be a nakanaka leaf before drying after harvest, or a nakanaka leaf prepared by processing the nakanaka leaf. The nakanaka leaf water extract of the present invention is preferably produced by a production method comprising a step of steaming cocoon leaves, a step of twisting nakanaka leaves, a step of extracting nakanaka leaves with water, and a step of concentrating the extract. Prepared. That is, the nakanaka leaf used in the present invention may be a nakanaka tea leaf prepared by a method including a step of steaming the cocoon leaf and a step of twisting the nakanaka leaf.

  In the present invention, the steaming step for the cocoon leaves can be carried out by a method usually performed in the art using a commercially available steamer or autoclave. For example, it is possible to heat-treat the green leaves by spreading the green leaves on a net conveyor and passing it through a processing chamber filled with non-pressure steam supplied from a boiler. The steaming temperature is not particularly limited, but may be appropriately selected within the range of 90 to 120 ° C., preferably 95 to 110 ° C., more preferably 100 to 110 ° C., for example, depending on the size of Tochu Nakaba. Further, the steaming time can be appropriately selected in the range of 10 to 240 seconds, preferably 20 to 180 seconds, more preferably 20 to 120 seconds. Moreover, the vapor | steam amount to be used can be suitably selected, for example in the range of 200-70 L / min, Preferably it is 170-100 L / min. The treatment amount of the steamed leaves is not particularly limited depending on the moisture content of the fresh leaves, but is appropriately selected within the range of, for example, 3 to 10 kg / min, preferably 4 to 8 kg / min, more preferably 5 to 7 kg / min. sell. This steaming process makes it easier to keep the ingredients of the fresh-leaved leaves by inactivating the enzyme that changes the brown-colored leaves, and the softening of the fresh leaves makes it easier to perform the subsequent twisting process. It brings about effects such as.

  The twisting step in the present invention can be performed using, for example, a commercially available twisting machine, roughing machine or intermediate hammering machine. For example, as a commercially available twister, Terada Seisakusho Co., Ltd., a twister 60 kg type etc. can be used. By this step, the moisture in the Tochu leaf is uniformly prepared while removing excess moisture, and further, it is easy to extract the components unique to Tochu. Although this process can be performed under heating as necessary, it is preferably performed without heating. The time required for this step is not particularly limited, but may be appropriately selected within a range of, for example, 10 to 80 minutes, preferably 20 to 60 minutes, more preferably 25 to 30 minutes. The treatment amount of the cocoon twisted leaves is not particularly limited, but may be appropriately selected within the range of, for example, 25 to 40 kg, preferably 30 to 35 kg, more preferably 32 to 33 kg depending on the moisture content. The moisture content of the chunaka leaf obtained through this step is, for example, 25 to 40%, preferably 25 to 35%, more preferably 25 to 30% on a dry basis.

  The method for preparing the Tochu tea leaves may include a step of roasting the Tochu leaves after the twisting process. The step of roasting the Nakanaka leaves is not particularly limited, but can be performed using, for example, a commercially available roaster. Although the roasting method in this process is not particularly limited, for example, it can be carried out by a hot air rotary drying fired machine manufactured by Yokoyama Seisakusho Co., Ltd. The time required for this step is not particularly limited, but may be appropriately selected within the range of 30 to 50 minutes, preferably 30 to 45 minutes, more preferably 35 to 40 minutes. The roasting temperature in this step is not particularly limited, but may be appropriately selected within the range of, for example, 100 to 140 ° C, preferably 120 to 140 ° C, more preferably 130 to 140 ° C. The water content of the chunaka leaf obtained through this step is, for example, 8% or less, preferably 4% or less, more preferably 2% or less on a dry basis.

  In the step of obtaining the water extract of Tochu leaf of the present invention, an amount of water appropriately selected from, for example, 5 to 50 kg, preferably 10 to 30 kg, more preferably 15 to 20 kg is used per 1 kg of Tochu tea leaves. it can. The water extraction of the Tochu leaf may be, for example, extraction with hot water at 85 to 105 ° C., preferably 90 to 95 ° C., or the water containing the ground material of Tochu leaf is cooled to a lower temperature (for example, 25 to 50). C., preferably 30 to 45.degree. C.).

  As used herein, “vascular endothelium-derived hyperpolarizing factor” means a factor that induces an endothelial cell-dependent hyperpolarizing reaction in smooth muscle cells induced in a state in which the production of nitric oxide and prostacyclin is inhibited. . Although the vascular endothelial hyperpolarizing factor has not been specifically identified at present, any chemical substance may be used in the present specification, or an electrical signal generated by a potential change generated in the endothelial cells. Enhancement of the vascular endothelium-derived hyperpolarizing factor means enhancing the hyperpolarizing reaction of smooth muscle caused by the vascular endothelial hyperpolarizing factor.

  The vascular endothelium-derived hyperpolarizing factor enhancer of the present invention can be used as a vascular relaxant, and can be preferably used for the relaxation of resistance blood vessels such as arterioles. In addition, the vascular endothelium-derived hyperpolarizing factor enhancer of the present invention can be used for the treatment or prevention of cardiovascular diseases, and preferably can be used for the treatment or prevention of microcirculation disorders. Here, the microcirculation disorder includes, for example, arteriosclerosis, heart disease, brain disease, myocardial infarction and cerebral infarction.

  According to another aspect of the present invention, there is provided the aforementioned vascular endothelium-derived hyperpolarizing factor enhancer for use in the treatment or prevention of coldness. Here, the term “coldness” means a state in which other parts of the body do not feel cold at all, while only a specific part feels unpleasant coldness. It can be used for the prevention or treatment of dizziness, hot flashes, lower abdominal pain, neurological symptoms such as insomnia, and chapped and moist. According to another feature of this aspect of the present invention, the aforementioned vascular endothelium-derived hyperpolarizing factor enhancer is also provided for use in imparting cold resistance by maintaining body temperature.

According to still another aspect of the present invention, pharmaceutical compositions and food compositions containing a vascular endothelium-derived hyperpolarizing factor enhancer are provided.
The extract used in the present invention may be a fraction obtained from the extract. The fractionation method may be a method usually used in the technical field to which the present invention belongs, and examples of the fractionation method include fractionation by extraction using an arbitrary solvent, gel filtration column chromatography, and ion exchange column. Fractions used, silica gel column chromatography and the like are included. The fractionation method may be one type of method or a combination of a plurality of means. Although it does not specifically limit as a solvent used by said fractionation method, For example, water, methanol, ethanol, n-propanol, isopropanol, acetic acid, acetone, and mixtures thereof can be used.

  The extract and fraction used in the present invention can be used as a concentrate, a viscous substance or a solid obtained by concentration and / or lyophilization, if necessary.

  The vascular endothelium-derived hyperpolarizing factor enhancer of the present invention can be used as an active ingredient of a pharmaceutical composition. The pharmaceutical composition can be used in various dosage forms such as tablets, capsules, powders, granules, pills, solutions, emulsions, suspensions, solutions, spirits, syrups, for oral administration. Extracts and elixirs can be used, and parenterals include, for example, injections such as subcutaneous injections, intravenous injections, intramuscular injections, intraperitoneal injections. Is not limited. The pharmaceutical composition of the present invention is preferably administered orally. These preparations can be produced by known methods usually used in the preparation process.

  The pharmaceutical composition may contain various commonly used components, such as one or more pharmaceutically acceptable excipients, disintegrants, diluents, lubricants, flavoring agents, Coloring agents, sweetening agents, flavoring agents, suspending agents, wetting agents, emulsifying agents, dispersing agents, adjuvants, preservatives, buffering agents, binders, stabilizers, coating agents and the like can be included. The pharmaceutical composition of the present invention may be in a sustained or sustained release dosage form.

  The dosage of the pharmaceutical composition of the present invention can be appropriately selected depending on the administration route, the patient's body shape, age, physical condition, degree of disease, elapsed time after onset, etc. An effective and / or prophylactically effective amount of a vascular endothelium-derived hyperpolarizing factor enhancer can be included. Tochu Nakaba was originally used as a Tochu tea leaf for serving beverages, and the Tochu Nakaba water extract is considered to be a relatively safe substance, so it can be administered at high concentrations as needed. Is possible. In the present invention, the Tochu tea leaf extract can generally be used as a dry amount of 1 to 8 g / day / adult, preferably 4.8 to 8 g / day / adult. Administration of the pharmaceutical composition may be a single dose or multiple doses, for example, ACE inhibitors, Ca antagonists, antiplatelet agents, coronary artery dilators, alpha blockers, beta blockers, angiotensin II receptor It can also be used in combination with other antihypertensive agents such as body antagonists and diuretics.

  According to still another aspect of the present invention, a food composition comprising the vascular endothelium-derived hyperpolarizing factor enhancer is provided. The food composition of the present invention includes a liquid beverage such as a functional beverage. In addition to being used as a functional food, the food composition can be used as a quasi-drug, a component such as food and drink, a food additive, and the like. In addition, the food composition in the present specification can be used as a functional food as it is, and can also be used as a component of food and drink, pharmaceuticals, quasi drugs, food and drink, food additives, and the like. The use enables daily and continuous ingestion of food and drink, food composition or composition for oral consumption having the effect of enhancing the hyperpolarization factor of vascular endothelium and the effect of improving the circulation of arterioles and resistance blood vessels associated therewith, It is possible to effectively improve the constitution by the effect, to treat and prevent diseases such as cardiovascular diseases, in particular, microcirculatory disorders and cold symptoms. Examples of food compositions, foods or beverages of the present invention include functional foods, health foods, and general foods (juices, confectionery) having a vascular endothelium-derived hyperpolarizing factor enhancing effect and an accompanying arteriole and resistance vascular circulation improving effect. , Processed foods, etc.), nutritional supplements (nutrient drinks, etc.). The food or beverage in the present specification includes, but is not limited to, inorganic components such as iron and calcium, various vitamins, dietary fiber such as oligosaccharide and chitosan, protein such as soybean extract, lipid such as lecithin, sucrose And saccharides such as lactose, plant extracts such as shiitake mushrooms, and the like.

  As shown in the following Examples, the vascular endothelium-derived hyperpolarizing factor enhancer of the present invention enhances the endothelium-dependent hyperpolarization reaction in smooth muscle cells, and has a relaxing action specific to blood vessels with a small diameter. Therefore, the present invention provides an effective means for treating and / or preventing cardiovascular diseases, particularly microcirculatory disorders and cold symptoms, which have a low burden on patients.

EXAMPLES Hereinafter, although the preferable Example of this invention is described in detail, this invention is not limited to these Examples.
[Example 1] Preparation of Tochu Naka leaf water extract (1-1) Manufacture of Tochu tea leaves Manufacture of Tochu tea leaves was performed based on the description in Example 2 of JP-A-8-173110. 5 kg of fresh leaves of Tochu were steamed at 110 ° C. for 90 seconds with a zonal steamer for producing Japanese tea. Fresh leaves were put into the machine from the inlet of the zonal steamer, steam was applied from the upper and lower steam supply devices while moving on the conveyor, and steamed at 110 ° C. for 90 seconds. By spreading the cocoon leaves on the net conveyor and passing them through a treatment chamber filled with non-pressure steam supplied from a boiler, the cocoon leaves can be steamed. For example, Miyamura Tekko Co., Ltd., a feeder, a ground type 1500, a net conveyor, and a banding type 1000 can be used.

  Next, after steaming, the steamed rice leaves were twisted for 30 minutes using a twisting machine, and the twisted material was dried using a dryer at 80 ° C. for 5 hours to a moisture content of 5%. The color of Tochu Nakaha, which was greenish brown after steaming, changed to greenish brown with drying. Then, it was roasted at 110 ° C. for 30 minutes using a fried leaf machine (IR-10SP type: Terada Seisakusho) to obtain 2 kg of Tochu tea leaves.

(1-2) Preparation of Tochu leaf extract 1 kg of Tochu tea leaf was put into 15 kg of hot water at 90 ° C. and extracted at 90 ° C. for 30 minutes to obtain 14 kg. Thereafter, the mixture was filtered using a 150 mesh filter, and the filtrate was cooled to 5 ° C. and left overnight. The supernatant was taken out, and the filtrate was concentrated at 50 ° C. under reduced pressure to obtain 1 kg.

The concentrated solution was processed with a centrifuge KS8000 manufactured by Kubota Corporation, the precipitate was removed by centrifugation at a rotational speed of 1800 rpm, and the resulting supernatant was sterilized by heating (85 ° C., 2 hours). An extract was obtained. The extract was dried by a spray drying method to obtain a powder (300 g) of Tochu Nakaba water extract as a brown powder.
[Example 2] Measurement of vasorelaxant action of Tochu leaf extract (2-1) Experimental procedure
Perfusion specimens of mesenteric arteries isolated from 8-10 week old Wistar male rats were prepared. After the specimen was placed in a perfusion apparatus kept at 37 ° C., Krebs solution was perfused at a constant flow rate, and the change in perfusion pressure was measured as a change in vascular tone with a pressure transducer TP-200T (Nihon Kohden). The outline of perfusion pressure measurement is shown in FIG. The perfusion device consists of a liquid pump (Peristaltic Pump AC-2120, ATTO), a pressure transducer (TP-200T, Nihon Kohden), an infusion pump (HARVARD975, USA), and a blood pressure amplifier (AP-641G, Nihon Kohden). . After attaching the specimen to the perfusion apparatus, Krebs solution is passed through a glass tube heated to 37 ° C. and heated, and the specimen is perfused at a constant flow rate of 5 mL / min. Changes in perfusion pressure are measured as vascular tone by a pressure transducer connected between the pump and the specimen.

The Krebs solution used in this experiment was prepared as follows. That is, 0.112 g of EDTA · 2Na (Dojindo Laboratories), 1.634 g of potassium dihydrogen phosphate (KH ₂ PO ₄ , Wako Pure Chemical), 2.958 of magnesium sulfate (MgSO ₄ · 7H ₂ O, Wako Pure Chemical) g, 69.544g of sodium chloride (NaCl, Wako Pure Chemical), 3.504g of potassium chloride (KCl, Wako Pure Chemical), 21.003g of sodium hydrogen carbonate (NaHCO ₃ , Wako Pure Chemical), weighed purified water (Wako Pure Chemical) The solution obtained by dissolving and diluting in 10) to make a total volume of 10 L was designated as Krebs 1 solution. Further, 17.66 g of calcium chloride dihydrate (CaCl ₂ .2H ₂ O, Wako Pure Chemical Industries, Ltd.) was weighed and dissolved and diluted with distilled water to adjust the total volume to 50 mL to obtain Krebs 2 solution. Next, 1 mL of Krebs 2 and 2 g of glucose (Wako Pure Chemical Industries, Ltd.) were dissolved and diluted with Krebs 1 solution aerated with 95% oxygen gas (Nippon Nitrogen) for 20 minutes to prepare a Krebs solution.

(2-2) Drugs used Acetylcholine hydrochloride (Daiichi Pharmaceutical), papaverine (Dainippon Pharmaceutical), methoxamine hydrochloride (Nippon Shinyaku), and sodium deoxycholate (Ishizu Pharmaceutical) were used.

(2-3) Measurement of vascular tone The Krebs solution was perfused into the mesenteric artery specimen under static tension, and then the blood vessel was contracted with methoxamine (7 μM) to increase the perfusion pressure to a certain level. Tochu liquid extract (100 pg / mL to 10 μg / mL) was dissolved in Krebs solution containing methoxamine to give a perfusate, which was perfused into the specimen to confirm the relaxation reaction. The magnitude of the vascular relaxation response is expressed as a relaxation rate with the relaxation response caused by papaverine (100 μM) perfusion as 100%.

(2-4) Removal of vascular endothelium Removal of vascular endothelial cells was performed by perfusing sodium deoxycholate (1.8 mg / mL) through a specimen in a quiescent state for about 30 seconds.

(2-5) Results The results of the relaxed state of the blood vessel in the presence of the vascular endothelium and the removal of the endothelial cells are shown in FIGS. 2 and 3, respectively. In the presence of vascular endothelium, relaxation of the blood vessels was observed by perfusing the Tochu leaf extract, but when the endothelial cells were removed, the relaxation reaction disappeared. From this, it was confirmed that the vasorelaxation reaction elicited by the Tochu Nakaha extract is dependent on endothelial cells.
[Example 3] Effect of additives on vasorelaxing action of Tochu leaf extract (3-1) Experimental procedure The perfusion pressure was measured by the same procedure as in Example 2. NO synthase inhibitor ^NG -nitro-L-arginine methyl ester (L-NAME, SIGMA CHEMICAL CO), cyclooxygenase inhibitor indomethacin (Wako Pure Chemical), depolarizer potassium chloride (KCl, Wako Pure Chemical) , Potassium channel inhibitor tetraethylammonium (TEA, Wako Pure Chemical), gap junction inhibitor 18α-glycyrrhetinic acid (18α-GA, Wako Pure Chemical), and antimuscarinic agent atropine (Wako Pure Chemical) Used as an agent. Add Krebs solution containing 2-7 μM methoxamine to L-NAME (100 μM), indomethacin (1 μM), KCl (high K ⁺ , 60 mM), TEA (5 mM), 18α-GA (10 μM) or atropine (1 μM) was dissolved and perfused, and the effect was observed.

(3-2) Results FIG. 4 shows the vascular relaxation reaction by the extract of Tochu leaf water when L-NAME, indomethacin and potassium chloride are added in the presence of the vascular endothelium. Addition of L-NAME (100 μM), a NO synthase inhibitor, enhanced the relaxation effect of the Tochu leaf extract. Addition of indomethacin (1 μM), a cyclooxygenase inhibitor, had little effect on relaxation. On the other hand, relaxation action was suppressed by adding potassium chloride (60 mM) as a depolarizing agent.

  FIG. 5 shows the vasorelaxation reaction with the Tochu leaf extract when TEA and 18α-GA are added in the presence of the vascular endothelium. Relaxation by the extract of Tochu leaf water by addition of TEA, a potassium channel inhibitor (5 mM), and addition of 18α-glycyrrhetinic acid, an inhibitor of gap junction that has been suggested to be involved in EDHF (10 μM) It was confirmed that the action was suppressed.

  Furthermore, FIG. 6 shows the vascular relaxation reaction by the Tochu leaf extract when atropine is added in the presence of vascular endothelium. The addition of atropine (1 μM), an antimuscarinic agent, inhibited relaxation only at high concentrations of Tochu Nakaba. From this, it was confirmed that the contribution rate of the acetylcholine receptor in the vasorelaxant action by the Tochu water extract was very low.

From the above results, it was confirmed that the vasorelaxant action by the Tochu water extract is mediated by EDHF.
[Example 4] Room temperature cooling load test In order to confirm the effect of the present invention on coldness, the test was performed according to the following procedure. 8 weeks old female Wistar rats (n = 6) were orally administrated with a foliage extract (1000 mg / kg) once a day for 9 days. Before administration, and after administration of the test substance on the first, third, fifth, seventh, and ninth days of administration, the animal is left in a laboratory at room temperature of 18 ° C., and immediately after that, the rectal temperature of each individual is adjusted at 20-26 ° C. It was measured. The rectal temperature after the start of the cooling test was compared with the rectal temperature before the start of the test. The results are shown in Table 1 and FIG.

In the vehicle control group, the low value after the start of administration changed with respect to the rectal temperature before the start of the test. In the Tochu Nakaha extract extract group, there was no rapid temperature drop due to room temperature cooling load, and the value was higher than that in the control group. From this, it was confirmed that the Tochu Nakaha water extract suppresses a decrease in body temperature due to room temperature cooling.
[Example 5] Water immersion cooling test In order to confirm the effect of the present invention on coldness, a test was performed according to the following procedure. The 8 weeks old Wistar female rats (n = 6) were orally administered with the Tochu leaf extract once a day for 9 days. After the final administration, after fasting for 24 hours, rectal temperature, footpad temperature, and body surface temperature were measured in a laboratory at room temperature of 20 ° C. (value before water immersion). After the measurement, the nakanaka leaf water extract and the medium were orally administered, and 30 minutes after the administration, they were immersed in water at 15 ° C. for 15 minutes using a stainless steel immersion cage. Immediately after the immersion, rectal temperature, footpad temperature and body surface temperature were measured. The body surface temperature was measured using a medical thermography device (Thermo Pure JTG, JEOL Ltd.). The temperature after water immersion (0 to 120 minutes) was compared with the value before water immersion. The results are shown in Tables 2 to 4 and FIGS.

  In the control group, a drastic decrease in body temperature was observed due to the water immersion cooling load, but gradually showed a recovery trend. In the Tochu Nakaha extract extract group, the body temperature decreased rapidly as in the control group due to the water immersion cooling load, but the subsequent recovery of body temperature was quick. From this, it was suggested that the Tochu Nakaha extract promotes the recovery of body temperature after water immersion cooling.

It is the schematic which showed the perfusion apparatus schematic diagram. It is a graph which shows transition of the relaxation reaction at the time of vascular endothelium presence when perfusion sample of the mesenteric artery extracted from the 8-10 week-old Wistar male rat perfusate the pericardium water extract. It is a graph which shows transition of the relaxation reaction at the time of vascular endothelium removal when perfusion specimen of the mesenteric artery extracted from 8-10 weeks old Wistar strain male rats is perfused with urchin extract. Tochu leaf extract when perfused specimen of mesenteric artery including vascular endothelium removed from 8-10 week old male Wistar rats was supplemented with NO synthase inhibitor, cyclooxygenase inhibitor or depolarizing agent It is a graph which shows transition of the relaxation reaction by. Of perfusion specimens of mesenteric arteries including vascular endothelium extracted from 8-10 week old Wistar male rats, and the relaxation reaction by the extract of Tochu leaf extract when perfused with potassium channel inhibitor and gap junction inhibitor It is a graph which shows transition. This is a graph showing the transition of relaxation response by the extract of Tochu leaf water when perfusion specimen of mesenteric artery including vascular endothelium extracted from 8-10 week old Wistar rats was perfused with antimuscarinic agent. is there. It is a graph which shows transition of the rectal temperature of the mouse | mouth which administered the Tochu water extract in the room temperature cooling addition test. It is a graph which shows transition of the rectal temperature after water immersion cooling about the mouse | mouth which administered the Tochu water extract. It is a graph which shows transition of the footpad temperature after water immersion cooling about the mouse | mouth which administered the Tochu water extract. It is a graph which shows transition of the body surface temperature after water immersion cooling about the mouse | mouth which administered the Tochu water extract.

Claims (10)

  Vascular endothelium-derived hyperpolarizing factor potentiator comprising urchin leaf extract.   The nakanaka leaf water extract is prepared by a manufacturing method comprising the steps of steaming cocoon leaf, twisting nakanaka leaf, extracting nakanaka leaf with water, and concentrating the extract. 2. The vascular endothelium-derived hyperpolarizing factor enhancer according to 1.   The vascular endothelium-derived hyperpolarizing factor enhancer according to claim 1 or 2 for use in relaxation of a resistance vessel.   The vascular endothelium-derived hyperpolarizing factor enhancer according to any one of claims 1 to 3, for use in the treatment or prevention of microcirculatory disorders.   The vascular endothelium-derived hyperpolarizing factor enhancer according to claim 4, wherein the microcirculation disorder is arteriosclerosis, heart disease, brain disease, myocardial infarction and cerebral infarction.   The vascular endothelium-derived hyperpolarizing factor enhancer according to any one of claims 1 to 3, for use in the treatment or prevention of coldness.   The vascular endothelium-derived hyperpolarizing factor enhancer according to any one of claims 1 to 3, for use in imparting cold resistance by maintaining body temperature.   A pharmaceutical composition comprising the vascular endothelium-derived hyperpolarizing factor enhancer according to any one of claims 1 to 7.   9. A pharmaceutical composition according to claim 8 for use in oral administration.   A food composition comprising the vascular endothelium-derived hyperpolarizing factor enhancer according to any one of claims 1 to 7.

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