CN115536522A - Use of pachyman cortex extract, pachyman neo-acid A and pachyman neo-acid B for regulating blood sugar - Google Patents

Abstract

Use of at least one of pachymic acid A and pachymic acid B for the preparation of a medicament or a food for regulating blood glucose. At least one of pachymic acid A and pachymic acid B may be used in the form of a plant extract, wherein the total content of pachymic acid A and pachymic acid B in the plant extract is not less than 30% by weight based on the total weight of the plant extract. The food can be health food, nutritional supplement food or special nutritional food.

Description

Use of pachyman cortex extract, pachyman neo-acid A and pachyman neo-acid B for regulating blood sugar

The application is a divisional application, the application date of the original application is 09.05.2017, and the application number is 201710320602.9, and the invention and creation names are 'the application of the poria peel extract, the pachyman neo-acid A and the pachyman neo-acid B in regulating blood sugar'.

Technical Field

The present invention relates to the use of extracts of poria peel, pachymic acid a and pachymic acid B, and in particular to the use of these substances for regulating blood glucose.

Background

Diabetes mellitus is a chronic metabolic disorder disease, and the main disease is that the glucose content in blood is too high due to abnormal operation of a mechanism for taking up glucose by cells in an organism. Generally, insulin secreted by pancreatic beta cells stimulates glucose uptake by adipocytes and muscle cells, and has the effect of regulating blood glucose. When an organism has insufficient insulin secretion or poor insulin sensitivity due to obesity, aging, etc., the blood glucose level is increased. Hyperglycemia may cause complications such as hypertension, heart disease, arteriosclerosis, hyperlipidemia, etc., and in severe cases, sequelae such as blindness, impotence, amputation, kidney washing, etc. may occur.

The current clinical treatment modes for diabetes mainly comprise modes of exercise, diet control, drug therapy and the like, wherein the drug therapy comprises insulin injection, oral hypoglycemic drugs such as sulfonylurea drugs (sulfonylureas), biguanide drugs (biguanidines), alpha-glucosidase inhibitors (alpha-glucosidase inhibitors), insulin sensitizers (insulin sensitizers) and the like. However, as human life forms change, the prevalence of diabetes also increases year by year. According to the prediction of the world health organization in 2008, it is expected that in 2030, diabetics will have more than 3 billion people in the mouth, and therefore, the industry is still working on developing a drug or a method which has low side effects and can effectively lower blood sugar.

The inventor of the present application has found that the poria peel extract and the contained pachymic acid a and pachymic acid B can effectively improve the glucose uptake ability of cells, and thus can be used for regulating blood sugar, especially providing excellent blood sugar lowering effect.

Disclosure of Invention

An object of the present invention is to provide a use of at least one of pachymic acid A and pachymic acid B for manufacturing a medicament or food for regulating blood glucose. The medicine or food is administered in an amount of about 0.05 mg/kg to 1 mg/kg body weight per day based on the total weight of pachymic acid A and pachymic acid B, and the food can be health food, nutritional supplement food or special nutritional food.

Preferably, at least one of pachymic acid A and pachymic acid B is used in the form of a plant extract. In the plant extract, the total content of pachymic acid a and pachymic acid B is not less than 30% by weight, preferably not less than 40% by weight, based on the total weight of the plant extract. More preferably, at least one of pachymic acid A and pachymic acid B is used in the form of extract of pachyma cocos peel. In the poria peel extract, the contents of pachymic acid (pachymic acid), dehydropachymic acid (dehydropachymic acid), temustic acid (temulosic acid) and dehydrotemustic acid (dehydrotemulosic acid) are not more than 0.5% by weight, and the contents of dehydrotrameteic acid (dehydrotrametinic acid), trameteic acid (trametinic acid), dehydrophellinus acid (dehydroeburicoic acid) and phellinus acid (eburicoic acid) are not more than 5% by weight, based on the total weight of the poria peel extract. Preferably, the content of dehydro-tetanic acid, dehydro-poronic acid and fomitonic acid in the poria peel extract is not more than 2.5% by weight based on the total weight of the poria peel extract. More preferably, the content of dehydro-streptococcic acid, dehydro-porococcic acid and fomes acid in the poria peel extract is not more than 1% based on the total weight of the poria peel extract.

It is another object of the present invention to provide a method for regulating blood glucose in a subject, comprising administering to a subject in need thereof an effective amount of at least one of pachymic acid a and pachymic acid B.

It is still another object of the present invention to provide a composition for regulating blood glucose, which is a medicament or food and comprises an effective amount of at least one of pachymic acid a and pachymic acid B.

Preferably, the extract of the skin part of poria cocos wolf is provided by an operation comprising the steps of:

(a) Extracting a skin part of poria cocos with a first solvent to obtain a crude extract;

(b) Drying the crude extract to obtain a crude extract powder;

(c) Extracting the crude extract powder with a second solvent to obtain a Poria peel extract,

wherein the first solvent and the second solvent are the same or different and are respectively selected from water, ethanol, alkali liquor, acid liquor and the combination of the above.

Wherein, the first solvent and the second solvent are ethanol water solutions with the same or different ethanol concentrations.

The invention has the beneficial effects that: the poria peel extract, the pachyman neo-acid A and the pachyman neo-acid B can really improve the capability of organism cells to take blood sugar, can be used for regulating blood sugar and particularly can reduce overhigh blood sugar.

Drawings

The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.

FIG. 1 shows the results of the effect of the extract from the skin of Poria cocos on the uptake of glucose in the culture medium by the mouse muscle cells, as measured by glucose oxidase assay;

FIG. 2 shows the results of the glucose oxidase assay of the effect of the extract from the skin of Poria cocos on the uptake of glucose into the culture medium by the adipocytes of mice;

FIG. 3A shows the results of the effect of pachyman neo-acid A on the ability of mouse muscle cells to take up glucose in the culture medium, as measured by glucose oxidase;

FIG. 3B shows the results of the effect of pachyman neo-acid B on the ability of mouse muscle cells to take up glucose in the culture medium, as measured by glucose oxidase;

FIG. 4A shows the results of the effect of pachyman neo-acid A on the ability of mouse adipocytes to take up glucose in the culture medium, measured by glucose oxidase;

FIG. 4B shows the effect of pachymanic acid B on the glucose uptake capacity of adipocytes in culture medium of mice measured by glucose oxidase.

Detailed Description

Some specific embodiments of the present invention will be described below; the invention may, however, be embodied in many different forms without departing from the spirit thereof, and the scope of the invention should not be construed as limited to the aspects set forth herein. Furthermore, unless the context requires otherwise, the words "a," "an," "the," and similar referents used in the specification (especially in the claims) are to be construed to cover both the singular and the plural; by "effective amount" is meant an amount of a compound that, when administered to a subject, is effective to increase the ability of the subject's cells to take up glucose; by "individual" is meant a mammal, which may be a human or a non-human animal; by "regulating blood glucose" is meant changing the glucose concentration in the blood towards normal values; the unit "mg/kg body weight" refers to the amount of drug required for an individual per kg body weight.

The use of a range of values (e.g., 5 to 100) in this specification should be understood to also include all rational numbers within the range as well as ranges consisting of any rational number within the range, and thus all possible combinations of values between the lowest value and the highest value recited are included in the range of values used in this specification. In addition, when "about" is used herein before a numerical value, it substantially means within 20%, preferably within 10%, and more preferably within 5% of the stated numerical value.

As explained above, the major disease of diabetes is excessive glucose content in blood due to abnormal operation of glucose uptake mechanism of cells in the body. The inventor of the application finds that pachyman neo-acid A and pachyman neo-acid B can improve the glucose uptake capability of cells, so that the pachyman neo-acid A and the pachyman neo-acid B can be used for regulating blood sugar, and especially reducing overhigh blood sugar. Therefore, the present invention provides an application of at least one of pachymanovanoic acid A and pachymaneoic acid B in regulating blood sugar, comprising the application of at least one of pachymaneoic acid A and pachymaneoic acid B in preparing a medicament or food for regulating blood sugar, a method for regulating blood sugar by administering at least one of pachymaneoic acid A and pachymaneoic acid B to an individual in need thereof, and a food or pharmaceutical composition comprising at least one of pachymaneoic acid A and pachymaneoic acid B.

Poria cocos (Schw.) Wolf is dry sclerotium of fungus (Poria cos (Schw.) Wolf) belonging to family Paralichoraceae. The Poria cocos fungi are often parasitic on the pine roots, with the outer skin being light brown or dark brown (poria peel), and the interior being pink or white (poria flesh). According to the classical record of traditional Chinese medicine, the flesh part of tuckahoe has the functions of calming, promoting urination, supplementing nutrition, enhancing immunity, delaying aging and the like, while the skin part of tuckahoe is only used for treating skin edema.

As shown in the following examples, according to the present invention, a poria peel extract containing not less than 30% by weight of pachymic acid A and pachymic acid B, based on the total weight of the extract, can be obtained from the skin of poria. Therefore, at least one of pachymic acid a and pachymic acid B used according to the present invention may be used in the form of a plant extract such as a pachyman cortex extract, wherein the total content of pachymic acid a and pachymic acid B in the plant extract used according to the present invention is not less than 30% by weight, preferably not less than 40% by weight, based on the total weight of the plant extract.

According to the present invention, the extract of the skin part of poria cocos employed may be an extract provided in an operation comprising the steps of: (a) Extracting the skin of Poria cocos with a first polar solvent to obtain a crude extract; (b) drying the crude extract to obtain a crude extract powder; (c) Extracting the crude extract powder with a second polar solvent to obtain a poria peel extract, wherein the first polar solvent and the second polar solvent are the same or different and are selected from water, ethanol, alkaline solution, acid solution, and combinations thereof. Wherein an alkaline solution refers to any suitable alkaline solution having a pH greater than 7 (e.g., sodium hydroxide solution), and an acidic solution refers to any suitable acidic solution having a pH less than 7 (e.g., hydrochloric acid solution). In some embodiments of the present invention, the first polar solvent and the second polar solvent are ethanol aqueous solutions with the same or different ethanol concentrations.

In the step (a), the ratio of the first polar solvent to the poria peel portion may be adjusted as required. In general, the amount of the first polar solvent is not particularly limited as long as the raw materials can be uniformly dispersed. For example, a volume ratio of the first polar solvent to the poria peel portion of about 8:1 to about 16:1, in an amount of 1. In one embodiment of the present invention, an ethanol aqueous solution is used as the first polar solvent, and the volume ratio of the ethanol aqueous solution is about 1:8, poria peel: the aqueous ethanol solution is subjected to the extraction of step (a).

In step (a), the extraction time may be suitably selected depending on the first polar solvent used. Using ethanol water solution as a first polar solvent, wherein the weight ratio of tuckahoe peel: the volume ratio of the ethanol water solution is 1: for example, the extraction is usually carried out for at least 1 hour, preferably at least 2 hours, more preferably at least 3 hours. Further, step (a) may be carried out by optionally carrying out other operations such as decoction, cooling, filtration, concentration under reduced pressure, chromatography on a resin column, and the like. Optionally, before step (a), the poria peel part is pre-soaked in the first polar solvent for a certain period of time. Taking the ethanol aqueous solution as the first polar solvent, the pre-soaking can be performed for about 12 hours.

In step (c), the ratio of the amount of the second polar solvent to the crude extract powder obtained from step (b) may be adjusted as desired. In general, the amount of the second polar solvent used is not particularly limited as long as the coarse extract powder can be uniformly dispersed. For example, a volume ratio of the second polar solvent to the poria peel extract powder of about 8:1 to about 16:1, in an amount of 1. In one embodiment of the present invention, an aqueous ethanol solution is used as the second polar solvent, and the volume ratio of the ethanol solution is about 1:8, powder of poria peel crude extract: extracting the aqueous solution of ethanol in the step (c).

The extract of the skin part of Poria cocos employed according to the present invention may also be a dried product, which may be provided by drying the extract obtained in step (c). Optionally, before step (b), repeating the extraction of the poria peel part with the same or different first polar solvent, and combining the extracts obtained from the multiple extractions to provide a crude extract for step (b); the cycle of step (b), step (c), and optionally other operations described above may also be repeated.

In the poria peel extract used according to the present invention, the total content of pachymic acid a and pachymic acid B is not less than 30% by weight, preferably not less than 40% by weight, based on the total weight of the poria peel extract, and each content of pachymic acid, dehydropachymic acid, temmoic acid, and dehydrotemmoic acid is not more than 0.5%, and each content of dehydrotetanic acid, tetanic acid, dehydrolaminic acid, and laminic acid is not more than 5%. Preferably, the content of dehydro-tetanic acid, dehydro-poronic acid and fomitonic acid in the poria peel extract is not more than 2.5% by weight based on the total weight of the poria peel extract. More preferably, the content of dehydro-streptococcic acid, dehydro-porococcic acid and fomes acid in the poria peel extract is not more than 1% based on the total weight of the poria peel extract.

The pharmaceutical composition or medicament provided for use according to the present invention may be in any suitable form, without particular limitation, in a corresponding suitable dosage form depending on the intended use. For example, but not limited thereto, the pharmaceutical composition or medicament may be administered to a subject in need thereof by oral or parenteral (e.g., subcutaneous, intravenous, intramuscular, or intraperitoneal) administration. Depending on the form and use, suitable carriers may be used to provide the pharmaceutical composition or medicament, wherein the carriers include excipients, diluents, adjuvants, stabilizers, absorption retarders, disintegrants, solubilizers, emulsifiers, antioxidants, binders, tackifiers, dispersants, suspending agents, lubricants, hygroscopic agents, etc.

In dosage forms suitable for oral administration, the pharmaceutical compositions or medicaments provided in the use according to the invention may contain any pharmaceutically acceptable carrier which does not adversely affect the desired benefit of the active ingredient (i.e., pachymic acid a, pachymic acid B, or pachyma glabra cortex extract), for example: water, saline solution, dextrose (dextrose), glycerol, ethanol or the like, cellulose, starch, sugar bentonite (sugar bentonite), and combinations of the foregoing. The pharmaceutical composition or medicament may be provided in a dosage form suitable for oral administration using any suitable method, for example: lozenges (e.g., dragees), pills, capsules, granules, powders, fluid extracts, solutions, syrups, suspensions, tinctures, and the like.

For injections or drops suitable for subcutaneous, intravenous, intramuscular, or intraperitoneal injection, the pharmaceutical composition or medicament provided by the application of the present invention may contain one or more components such as isotonic solution, salt buffer (e.g., phosphate buffer or citrate buffer), solubilizer, emulsifier, 5% sugar solution, and other carriers, and may be provided in dosage forms such as intravenous infusion solution, emulsion intravenous infusion solution, dry powder injection solution, suspension injection solution, or dry powder suspension injection solution. Alternatively, the pharmaceutical composition or medicament may be prepared as a pre-injection solid, provided in a dosage form that is soluble in other solutions or suspensions, or in an emulsifiable dosage form, and dissolved or emulsified in other solutions or suspensions prior to administration to a subject in need thereof to provide the desired injection.

If necessary, the pharmaceutical composition or medicament provided by the use according to the present invention may further contain additives in appropriate amounts, such as flavors, toners, colorants, and the like, which can improve the mouth-feel and visual sensation of the pharmaceutical composition or medicament upon administration, and buffers, preservatives, antibacterial agents, antifungal agents, and the like, which can improve the stability and storability of the medicament. In addition, the pharmaceutical composition or medicament may optionally further comprise one or more other active ingredients (e.g., antioxidants, insulin sensitizers, etc.), or may be used in combination with the medicament comprising the one or more other active ingredients, to further enhance the efficacy of the pharmaceutical composition or medicament or to increase the flexibility and formulation of the formulation, as long as the other active ingredients do not adversely affect the efficacy of the active ingredient of the present invention (i.e., pachymic acid a, pachymic acid B, or the extract of the skin of poria).

The pharmaceutical compositions or medicaments provided for use according to the present invention can be administered at different dosing frequencies, once a day, multiple times a day, or once a few days, depending on the need, age, weight, and health condition of the subject to be administered. For example, when administered orally to a subject to regulate blood glucose, the amount is about 0.01 mg/kg to 5 mg/kg body weight per day, preferably about 0.03 mg/kg to 2 mg/kg body weight per day, more preferably about 0.05 mg/kg to 1 mg/kg body weight per day, based on the total weight of pachymic acid a and pachymic acid B. Alternatively, the amount of poria peel extract is from about 0.025 mg/kg to about 25 mg/kg body weight per day, preferably from about 0.075 mg/kg to about 10 mg/kg body weight per day, and more preferably from about 0.125 mg/kg to about 5 mg/kg body weight per day.

The food provided by the application of the invention can be health food, nutrition supplement food or special nutrition food, and can be made into products such as dairy products, meat processed products, breads, flour food, biscuits, buccal tablets, capsules, juices, teas, sports drinks, nutrition drinks and the like, but not limited thereto. Preferably, the food product for use according to the present invention is provided in the form of a health food.

The health food, nutritional supplement food and special nutritional food provided by the use according to the present invention can be taken at different frequencies such as once a day, multiple times a day, or once a few days, and the recommended intake varies depending on the age, weight and health condition of the subject to whom the food is administered. The contents of pachyman neo-acid a, pachyman neo-acid B, or a pachyman cortex extract in the health food, the nutritional supplement food, and the special nutritional food provided by the present invention can also be adjusted to a specific group, preferably to an amount to be taken daily. For example, if an individual has a recommended intake of about 70 mg total pachymic acid a and pachymic acid B per day based on the total weight of pachymic acid a and pachymic acid B, and the health food contains about 35 mg total pachymic acid a and pachymic acid B per serving, the individual may consume about two servings of the health food per day.

The recommended usage amount, the usage standard and condition of a specific group (such as pregnant women, diabetics and patients with kidney diseases) or the recommended matters for taking together with other foods or medicines can be marked on the outer package of the health food, the nutritional supplement food and/or the special nutritional food, so that the health food, the nutritional supplement food and/or the special nutritional food can be taken by a user at home without the guidance of doctors, pharmacists or related staff without safety concerns.

The present invention also provides a method of regulating blood glucose comprising administering to an individual in need thereof an effective amount of an active ingredient, wherein the active ingredient is at least one of pachymic acid a and pachymic acid B. In the method for regulating blood sugar according to the present invention, the state, administration route, administration form, applicable dose and application of the related treatment of the active ingredient are as described above.

The invention will now be further illustrated by the following examples. These examples are provided for illustration only and are not intended to limit the scope of the present invention. The scope of the invention is indicated in the claims.

Examples

[ preparation examples ]

A. Preparation of extract of Poria cortex

A-1, taking Poria (Yunnan, from Yunnan province), cleaning, peeling off its outer skin (hereinafter called "Poria peel part"), and taking the rest as the flesh part (hereinafter called "Poria flesh part"). Taking the poria peel part, and mixing the poria peel part at room temperature in a proportion of 1:8 (Poria cocos Wolff) in 75% ethanol aqueous solution for 12 hours, then boiled and extracted (for 3 hours). The previous extraction steps were repeated three times. The extracts obtained from the three extractions were combined and filtered to remove insoluble matter, to obtain a crude extract. Then, the crude extract is concentrated under reduced pressure to remove the solvent, and then dried by a spray dryer to obtain a crude extract powder.

A-2, taking the crude extract powder obtained in A-1, and mixing the crude extract powder with a solvent of 1:8 (crude extract powder: aqueous ethanol solution), and extracting with 95% ethanol for 3 hours, and separating with a column using silica gel as stationary phase to obtain a poria peel extract.

A-3. Taking the crude extract powder obtained from A-1, mixing the crude extract powder with 1:10 The volume ratio of (crude extract powder: water) was uniformly dispersed in pure water. Then, sodium hydroxide was added to raise the pH of the mixture to about 12, and then poured into a tempering tank maintained at 65 ℃ and stirred uniformly until the reaction was complete. Then, neutralization was carried out with 12N concentrated hydrochloric acid, the filtrate was removed after centrifugation, and the remaining insoluble matter was washed with pure water and dried with a spray dryer to obtain an extract powder. Taking the extract powder, and mixing the extract powder with a weight ratio of 1:20 (extract powder: 1N aqueous sodium bicarbonate solution) was extracted three times with 1N aqueous sodium bicarbonate solution, the extracts obtained by the three extractions were combined, neutralized with 12N concentrated hydrochloric acid, the filtrate was centrifuged, and the remaining insoluble matter was washed with pure water and dried by a spray dryer to obtain a poria peel extract.

A-4, detecting the components of the extracts obtained from A-2 and A-3 by liquid chromatography/ultraviolet light/mass spectrometry at the wavelengths of 243 nm and 210 nm, respectively, and quantifying the content of each component in each of the extracts by high performance liquid chromatography, wherein the analysis results of the extracts obtained from A-2 are shown in Table 1, and the analysis results of the extracts obtained from A-3 are shown in Table 2.

TABLE 1

TABLE 2

Composition (I)	By weight%
		Pachymic Acid (PA)	－
Dehydropachymic acid (DPA)	－
		Tumoranic Acid (TA)	－
Dehydrotemmonic acid (DTA)	0.31
		Polyporus acid C (Polyphonic acid C, PAC)	1.15
3-epi-dehydrotemmoic acid (3-epi-dehydrotumulosic acid, EDTA)	0.40
		Dehydroartemisinic acid (DTTA)	0.58
Trametenolic Acid (TTA)	0.35
		Poria cocos neo-acid A (poricoic acid A, PAA)	41.06
Dehydroabietic acid (DEA)	0.19
		Poria cocos neo-acid B (poricoic acid B, PAB)	16.50
Phellinus linteus acid (eburicoic acid, EA)	0.20

As can be seen from table 1, the poria peel fraction extract obtained in a-2 contained high amounts of pachymic acid a (32.72% by weight of the extract) and pachymic acid B (10.44% by weight of the extract), low amounts of dehydro-tetanic acid, dehydro-poronic acid and fomes acid (2.01%, 0.85%, 1.2%, 0.83% by weight of the extract, respectively), and very low amounts of dehydro-ramosic acid (0.46% by weight of the extract), but no pachymic acid, dehydro-pachymic acid and ramosic acid.

As shown in table 2, the poria peel extract obtained in a-3 contained high amounts of pachymic acid a (41.06% by weight of the extract) and pachymic acid B (16.50% by weight of the extract), low amounts of grignard acid C, dehydro-streptococcic acid (1.15% by weight and 0.58% by weight of the extract), and very low amounts of dehydro-streptococcic acid, 3-epi-dehydro-streptococcic acid, dehydro-streptococcic acid and mycolic acid (0.31%, 0.40%, 0.35%, 0.19%, 0.20% by weight of the extract), but did not contain pachymic acid, dehydro-streptococcic acid and oxytetracycline.

B. Preparation of pachyman neo-acid A and pachyman neo-acid B

B-1, 1:500 (poria peel extract: methanol) in such a volume ratio that the poria peel extract obtained in A-2 or A-3 is uniformly dissolved in methanol. Filtering to remove insoluble substances, separating and collecting pachymanic acid A and pachymanic acid B with preparative high-performance liquid chromatography (using methanol mixed with water as mobile phase) at wavelength of 243 nm, collecting, and removing methanol with vacuum concentrator to obtain pachymanic acid A and pachymanic acid B.

B-2, detecting the pachyman neo-acid A and the pachyman neo-acid B obtained by the B-1 by a liquid chromatography/ultraviolet light/mass spectrometer under the wavelength of 243 nm respectively, and the results show that the purities of the pachyman neo-acid A and the pachyman neo-acid B are both more than 98%.

C. Cell culture

Fully differentiated mouse muscle cells (C2C 12, from ATCC) and adipocytes (3T 3-L1, from ATCC) were each cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 2% Bovine Serum Albumin (BSA) but no serum (serum-free) for 16 hours. Next, the cells were washed with Dulbecco's phosphate-buffered saline (D-PBS), and the medium was replaced with 0.2% BSA-DMEM medium containing 500. Mu.g/ml glucose for subsequent experiments.

Example 1: effect of Poria cortex extract on glucose uptake ability of cells

(1-1) muscle cells

The C2C12 cells provided in [ preparation example ] were taken, divided into five groups and cultured in the following media for 2 hours, respectively:

1. group I: BSA-DMEM medium containing 500 micrograms of glucose/ml;

2. group II: BSA-DMEM medium containing 500 micrograms glucose/ml, and insulin at a concentration of 100 nemo;

3. group III: BSA-DMEM medium containing 500. Mu.g glucose/ml and the poria peel extract solution provided in [ preparation example A-2] at a concentration of 0.1. Mu.g/ml;

4. group IV: BSA-DMEM medium containing 500. Mu.g glucose/ml and the poria peel extract solution provided in [ preparation example A-2] at a concentration of 1. Mu.g/ml;

5. group V: BSA-DMEM medium containing 500. Mu.g glucose/ml and the poria peel extract solution provided in preparation example A-2 at a concentration of 10. Mu.g/ml.

Thereafter, the glucose content of each cell culture was measured by glucose oxidase method to determine the degree of glucose consumption (representing the ability of the cells to take up glucose) of each group. Finally, relative glucose uptake capacity was calculated for each of the other groups based on the results of the control group (i.e., cells cultured in group I broth), and the results are shown in FIG. 1.

As can be seen from fig. 1, the glucose uptake capacity of the cells treated with the poria peel extract of the present invention (i.e., cells cultured in group III, IV, or V culture broth) was significantly improved, even beyond that of the positive control group (i.e., cells cultured in group II culture broth), compared to the control group. The above results show that the extract of the skin part of poria cocos of the present invention can effectively enhance the glucose uptake ability of muscle cells, and thus can be used for regulating blood glucose.

(1-2) adipocytes

The 3T3-L1 cells provided in [ preparation example ] were taken, divided into five groups and cultured in group I to V media described in example 1 (1-1), respectively, for 2 hours. Then, glucose content in the culture solution of each group of cells was measured by glucose oxidase method to find out the degree of glucose consumption (representing the ability of the cells to take up glucose) of each group. Finally, relative glucose uptake capacity was calculated for each of the other groups based on the results of the control group (i.e., cells cultured in group I broth), and the results are shown in FIG. 2.

As can be seen from fig. 2, the glucose uptake capacity of the cells treated with the poria peel extract of the present invention (i.e., cells cultured in group III, IV, or V culture medium) tended to increase compared to the control group, with the increase in the high dose group (cells cultured in group V culture medium) being most pronounced. The results show that the poria peel extract of the invention can effectively improve the glucose uptake capability of fat cells, and thus can be used for regulating blood sugar.

Example 2: effect of pachyman neo-acid A and pachyman neo-acid B on glucose uptake ability of cells

(2-1) muscle cells

The C2C12 cells provided in [ preparation example ] were taken, divided into eight groups and cultured in the following media for 2 hours, respectively:

1. group i: BSA-DMEM medium containing 500 micrograms of glucose/ml;

2. group ii: BSA-DMEM medium containing 500 micrograms glucose/ml, and insulin at a concentration of 100 nemo;

3. group ii i: BSA-DMEM medium containing 500. Mu.g glucose/ml and pachymic acid A or pachymic acid B provided in [ preparation example B-1] at a concentration of 0.1. Mu.g/ml;

4. group iv: BSA-DMEM medium containing 500. Mu.g glucose/ml and pachymaneoic acid A (or pachymaneoic acid B) provided in preparation example B-1 at a concentration of 1. Mu.g/ml;

5. group v: BSA-DMEM medium containing 500. Mu.g/ml glucose and pachymic acid A (or pachymic acid B) provided in preparation example B-1 at a concentration of 10. Mu.g/ml.

Then, the glucose content in the culture solution of each group of cells was measured by glucose oxidase method to find the degree of glucose consumption (representing the ability of the cells to take up glucose) of each group, and finally, the relative glucose uptake ability of each other group was calculated based on the results of the control group (i.e., the cells cultured in the culture solution of the i-th group), and the results are shown in fig. 3A and 3B. Wherein, fig. 3A contains the results of control, positive control (i.e., cells cultured in group ii broth), and pachymic acid a treated group (i.e., cells cultured in group iii, iv, or v broth, wherein the broth contains pachymic acid a). FIG. 3B contains the results for control, positive control, and pachymic acid B treated groups (i.e., cells cultured in group iii, iv, or v media containing pachymic acid B).

As can be seen from fig. 3A, the glucose uptake capacity of the group treated with pachymosin a was significantly increased compared to the control group to be comparable to the positive control group. The results show that pachymic acid A can effectively improve the glucose uptake capability of muscle cells, and can be used for regulating blood sugar.

As can be seen from fig. 3B, the glucose uptake capacity of the group treated with pachymic acid B was also significantly increased compared to the control group. The foregoing results show that pachymic acid B can also regulate blood glucose by increasing the ability of muscle cells to take up glucose.

(2-2) adipocytes

The 3T3-L1 cells provided in [ preparation example ] were taken, divided into eight groups and cultured in the group i to v media described in example 2 (2-1), respectively, for 2 hours. Then, the glucose content of each cell culture fluid was measured by glucose oxidase method to find the glucose consumption degree (representing the ability of the cells to take up glucose) of each group. Finally, the relative glucose uptake capacity of each of the other groups was calculated based on the results of the control group (i.e., cells cultured in the culture broth of group i), and the results are shown in FIGS. 4A and 4B. Wherein, fig. 4A contains the results of control, positive control (i.e., cells cultured in group ii broth), and pachymic acid a treated group (i.e., cells cultured in group iii, iv, or v broth wherein the broth contains pachymic acid a). Fig. 4B contains the results for control, positive control, and pachymic acid B treated groups (i.e., cells cultured in group iii, iv, or v media containing pachymic acid B).

As can be seen from fig. 4A, the glucose uptake capacity of the pachymic acid a-treated group tended to increase compared to the control group, with the increasing tendency being significant in the medium dose group (i.e., cells cultured in the iv group culture medium) and the high dose group (i.e., cells cultured in the v group culture medium). The results show that pachymic acid A can effectively improve the glucose uptake capability of fat cells, and can be used for regulating blood sugar.

As can be seen from fig. 4B, the glucose uptake capacity of the group treated with pachymic acid B was significantly increased compared to the control group. The foregoing results show that pachymic acid B can also regulate blood glucose by increasing the ability of adipocytes to take up glucose.

As shown in the above examples, the poria peel extract, pachymic acid a and pachymic acid B of the present invention can indeed improve the ability of living cells to take up blood sugar, and can be used for regulating blood sugar, and especially can reduce too high blood sugar.

Claims (10)

1. A method for providing an extract from the skin portion of Poria cocos comprising the steps of:

(a) Extracting a skin part of poria cocos with a first solvent to obtain a crude extract;

(b) Drying the crude extract to obtain a crude extract powder; and

(c) Extracting the crude extract powder with a second solvent to obtain a Poria peel extract,

wherein the first solvent and the second solvent are the same or different and are selected from water, ethanol, alkali solution, acid solution, and combinations thereof.

2. The method of claim 1, wherein the first solvent and the second solvent are aqueous solutions of ethanol having the same or different ethanol concentrations.

3. The method of claim 1, wherein the first solvent is ethanol and the second solvent is a lye.

4. The method of claim 3, wherein step (c) is repeated with the same or different lye.

5. The method according to any one of claims 1 to 4, wherein before step (b), the extraction of the pericarp fraction of Poria cocos is repeated with the same or a different first solvent, and the extracts obtained from the multiple extractions are combined to provide the crude extract subjected to step (b).

6. An extract of the skin of Poria cocos provided by the method of claim 1, which contains pachymic acid A and pachymic acid B and is free of pachymic acid, dehydropachymic acid and temmoic acid.

7. The poria peel extract according to claim 6, wherein the total content of pachymic acid A and pachymic acid B is not less than 30% by weight based on the total weight of the poria peel extract.

8. The poria peel extract according to claim 6, wherein the dehydrotumaric acid is present in an amount of no more than 0.5% by weight, based on the total weight of the poria peel extract.

9. The poria peel extract according to claim 6, wherein the contents of dehydro-tetanic acid, dehydro-poronic acid and fomes acid are each not more than 5% by weight based on the total weight of the poria peel extract.

10. Use of the poria peel extract according to any one of claims 6 to 10 for the preparation of a medicament or food for regulating blood glucose.

Images