CN113694120A - Plant extract composition with weight-losing and lipid-lowering activities and preparation method and application thereof - Google Patents
Plant extract composition with weight-losing and lipid-lowering activities and preparation method and application thereof Download PDFInfo
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- CN113694120A CN113694120A CN202111224581.3A CN202111224581A CN113694120A CN 113694120 A CN113694120 A CN 113694120A CN 202111224581 A CN202111224581 A CN 202111224581A CN 113694120 A CN113694120 A CN 113694120A
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- plant extract
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- extract composition
- lipid
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/82—Theaceae (Tea family), e.g. camellia
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
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Abstract
The application relates to the field of medicines, and particularly discloses a plant extract composition with weight-losing and lipid-lowering activities, and a preparation method and application thereof. The preparation method of the plant extract composition with the weight-losing and lipid-lowering activity comprises the following steps: (1) crushing and degreasing Pu' er tea, grape skin, apple skin and peony to obtain a raw material mixture; (2) adding an organic solvent aqueous solution into the raw material mixture obtained in the step (1), mixing, performing ultrasonic extraction, and collecting an extracting solution to obtain a crude extracting solution; (3) and (3) concentrating the crude extract obtained in the step (2) under reduced pressure until no organic solvent smell exists, further concentrating until the solvent is volatilized, and drying to obtain the target extract. The plant extract composition can be used for preparing medicines, health-care foods, food additives and the like with weight-losing or anti-cancer effects, and has the advantages of high fatty acid synthase inhibition activity, good weight-losing and lipid-lowering effects and excellent tumor cell inhibition effect.
Description
Technical Field
The application relates to the technical field of medicines, in particular to a plant extract composition with weight-losing and lipid-lowering activities, and a preparation method and application thereof.
Background
Currently, the incidence of obesity is on the rise, both in developed and developing countries. Obesity is a group of common metabolic diseases, and when a person eats more calories than consumed calories, the excess calories are stored in the body in the form of fat, the amount of which exceeds the normal physiological requirement, and when a certain amount is reached, obesity develops.
Fatty Acid Synthase (FAS) is the most important enzyme in the synthesis of fat in biological cells. Studies of fatty acid synthase in poultry by Tianwei et al found that the fat level in abdominal cavity of poultry and the activity of fatty acid synthase had a very high positive correlation, and proposed the theory that "controlling the activity of fatty acid synthase is an effective method for controlling the fat level in animals" (Tianwei, 1994. regulation of body fat level and fatty acid synthase activity in animals. chemistry of life, 14, (1): 184; Tianwei et al, 1996. relationship of body fat level and liver fatty acid synthase activity in layers of different growth stages. J. Biochemistry, 12(2): 234-236; Mei Li et al, 1999. fans entering the levels of layers of fat acid synthase activity in poultry Mol. Biom Mol Biol Int, 47(1): 63-69). Results of studies on mouse fatty acid synthase by loftus et al, university of pockins, in 2000, demonstrated that inhibition of fatty acid synthase resulted in accumulation of malonyl-coa, which inhibited the expression of food-related hypothalamic neuropeptide Y, resulting in decreased appetite and weight loss without affecting the mobility of mice. In addition Loftus et al indicate that fatty acid synthase can be a potential therapeutic target for weight control (Loftus. T.M.et al, 2000.Science, 288(5475): 2379-.
In addition, many studies have found that the activity and expression of FAS is shown to be extremely low in almost all non-diseased adult tissues, while its activity is significantly increased and expression is significantly up-regulated in many types of cancer tissues, such as rectal cancer, breast cancer, prostate cancer, endometrial cancer, ovarian cancer, lung cancer, liver cancer, and the like. Meanwhile, it has been found that cerulenin, an inhibitor of FAS, can kill cancer cells and inhibit the growth of xenograft tumors, and other FAS inhibitors such as cerulenin derivative C75, antiobesity drug orlistat, EGCG in green tea and other flavonoids of natural origin such as luteolin, quercetin, kaempferol and the like and antibiotic triclosan have also been confirmed to inhibit the growth of tumor cells by inducing apoptosis.
The above results indicate that fatty acid synthase may be a potential dual target for the treatment of obesity and tumors. Therefore, the development and development of the fatty acid synthase inhibitor have great display significance and development potential.
Disclosure of Invention
In order to enhance the inhibition activity of fatty acid synthase and reduce the synthesis of fat, the application provides a plant extract composition with the activity of losing weight and reducing fat, a preparation method and application thereof.
In a first aspect, the present application provides a method for preparing a plant extract composition with weight-losing and lipid-lowering activities, which adopts the following technical scheme:
a method for preparing a plant extract composition with weight-losing and lipid-lowering activities comprises the following steps:
s1, mixing Pu' er tea, grape skin, apple skin and peony according to the weight ratio of 2-4:0.5-1.5:2-4:0.5-1, crushing and degreasing to obtain a raw material mixture;
s2, mixing the raw material mixture obtained in the step S1 with an organic solvent aqueous solution, carrying out ultrasonic extraction, and collecting an extracting solution to obtain a crude extracting solution;
s3, concentrating the crude extract obtained in the step S3 under reduced pressure until no organic solvent smell exists, further concentrating until the solvent is volatilized, and drying to obtain the target extract.
By adopting the technical scheme, the four raw materials of Pu' er tea, apple peel, grape peel and peony flower are used as the raw materials of the extract, and then the raw materials are subjected to degreasing, ultrasonic extraction and other procedures by controlling the dosage of the four raw materials, wherein the degreasing operation is to remove fatty acids and other fat-soluble components in the four plant raw materials, and the ultrasonic extraction is to complete the extraction of the effective components with the maximum efficiency under the relatively low temperature condition.
Preferably, in step S2, the concentration of the aqueous solution of organic solvent is 50-90% by volume.
By adopting the technical scheme, when the concentration of the organic solvent aqueous solution is low, the effective components in the plant cannot be completely extracted, and when the concentration of the organic solvent aqueous solution is high, the activity of the effective components of the plant extract is adversely affected.
Preferably, in the step S2, the feed-liquid ratio of the raw material mixture to the organic solvent aqueous solution is 1g (8-15) mL.
By adopting the technical scheme, the feed-liquid ratio is controlled to be 1g (8-15) mL, so that the extraction rate is high, and the weight loss and food inhibition effects are good.
Preferably, the organic solvent in step S2 is selected from one of formic acid, acetic acid, methanol, ethanol, n-propanol, isopropanol and acetone.
By adopting the technical scheme, the organic solvent such as ethanol and methanol is taken as the extracting solution, the extraction time is short, and the dissolved impurities are less.
Preferably, the ultrasound extraction in step S2 is specifically performed by: ultrasonic extracting at room temperature for 1-3 times, each time for 10-30 min.
By adopting the technical scheme, ultrasonic extraction is carried out at room temperature, the condition is mild, and chemical components in the extract cannot be damaged.
Preferably, in the step S3, the concentration under reduced pressure is carried out at a temperature of 30-60 ℃ and a pressure of 0.01-0.1 MPa.
By adopting the technical scheme, the concentration temperature is 30-60 ℃, the damage of certain effective components caused by overhigh temperature is prevented, and the increase of impurities is prevented.
Preferably, in step S1, the specific method of degreasing is: adding pulverized Pu her tea, grape peel, apple peel and peony pollen into petroleum ether or cyclohexane, heating and refluxing in water at 30-60 deg.C for 10-30min for 2-3 times, and filtering to remove petroleum ether or cyclohexane.
By adopting the technical scheme, cyclohexane or petroleum ether is used as a degreasing solvent, the fat-soluble components are completely dissolved, the water-soluble components are basically insoluble, the fatty acids and the fat-soluble impurities can be removed,
in a second aspect, the present application provides a plant extract composition with weight-reducing and lipid-lowering activities, which adopts the following technical scheme: is prepared by a preparation method of a plant extract composition with the activity of losing weight and reducing fat.
In a third aspect, the present application provides a fatty acid synthase inhibitor, which adopts the following technical scheme: comprises a plant extract composition with the activity of losing weight and reducing fat.
In summary, the present application has the following beneficial effects:
1. because the extract formula with the weight-losing activity is prepared from edible plants including Pu' er tea, apple peel, grape peel and peony flower, and the extract formula is a natural, plant-derived and non-toxic fatty acid synthase inhibitor, the sources of the natural fatty acid synthase inhibitor are enriched, and the new efficacy and application value of the composition are developed.
2. The plant extract composition prepared in the application can obviously inhibit the activity of fatty acid synthase, the activity of the plant extract composition is 1-10 times higher than the oxygen index of the currently known fatty acid synthase such as cerulenin, EGCG, resveratrol and the like, and the plant extract composition can reduce the weight and inhibit the appetite after being orally taken.
3. The application adopts plant resources with dual purposes of medicine and food, takes the Pu' er tea, grape skin, apple skin and peony prepared extract formula as raw materials to prepare health-care food for losing weight or resisting tumor and the like, and has wider application and development prospect.
Detailed Description
Examples
Example 1: a method for preparing a plant extract composition with weight-losing and lipid-lowering activities comprises the following steps:
s1, mixing and crushing Pu 'er tea, grape skin, apple peel and peony flower according to the weight ratio of 3:1:3:1, sieving with a 20-mesh sieve, adding the crushed substances into petroleum ether, heating and refluxing in water bath at 60 ℃ for 3 times, each time for 30min, filtering to remove the petroleum ether to obtain a raw material mixture, wherein the material-liquid ratio of the crushed substances of the Pu' er tea, the grape skin, the apple peel and the peony flower to the petroleum ether is 1g:15 mL;
s2, mixing the raw material mixture obtained in the step S1 with an organic solvent water solution with the volume percentage of 60% according to the material-liquid ratio of 1g:15mL, carrying out ultrasonic extraction for 3 times at room temperature with the power of 200W, keeping the ultrasonic extraction for 10min each time, collecting and combining the extracting solutions to obtain a crude extracting solution, wherein the organic solvent is ethanol;
s3, concentrating the crude extract obtained in the step S3 under reduced pressure at 50 ℃ and 0.05MPa until no ethanol smell exists, further concentrating until the solvent is volatilized, and drying until the weight is constant to obtain the plant extract composition.
Example 2: a method for preparing a plant extract composition with weight-losing and lipid-lowering activities comprises the following steps:
s1, mixing and crushing Pu 'er tea, grape skin, apple peel and peony flower according to the weight ratio of 3:1:3:1, sieving with a 20-mesh sieve, adding the crushed substances into petroleum ether, heating and refluxing in water bath at 30 ℃ for 3 times, filtering for 20min each time, removing the petroleum ether to obtain a raw material mixture, wherein the material-liquid ratio of the crushed substances of the Pu' er tea, the grape skin, the apple peel and the peony flower to the petroleum ether is 1g:10 mL;
s2, mixing the raw material mixture obtained in the step S1 with 70% by volume of organic solvent aqueous solution according to a material-liquid ratio of 1g:10mL, carrying out ultrasonic extraction for 2 times at room temperature with 200W power, keeping the ultrasonic extraction for 30min each time, collecting and combining the extracting solutions to obtain a crude extracting solution, wherein the organic solvent is methanol;
s3, concentrating the crude extract obtained in the step S3 under reduced pressure at 55 ℃ and 0.01MPa until no methanol smell exists, further concentrating until the solvent is volatilized, and drying until the weight is constant to obtain the plant extract composition.
Example 3: a method for preparing a plant extract composition with weight-losing and lipid-lowering activities comprises the following steps:
s1, mixing and crushing Pu 'er tea, grape skin, apple peel and peony flower according to the weight ratio of 3:1:3:1, sieving with a 20-mesh sieve, adding the crushed material into cyclohexane, heating and refluxing in water bath at 60 ℃ for 3 times, each time for 30min, filtering to remove cyclohexane to obtain a raw material mixture, wherein the material-liquid ratio of the crushed material of Pu' er tea, grape skin, apple peel and peony flower to cyclohexane is 1g:15 mL;
s2, mixing the raw material mixture obtained in the step S1 with 70% by volume of organic solvent aqueous solution according to a material-to-liquid ratio of 1g to 8mL, carrying out ultrasonic extraction at room temperature for 1 time at 200W for 30min each time, collecting and combining the extracting solutions to obtain a crude extracting solution, wherein the organic solvent is methanol;
s3, concentrating the crude extract obtained in the step S3 under reduced pressure at 30 ℃ and 0.1MPa until no methanol smell exists, further concentrating until the solvent is volatilized, and drying until the weight is constant to obtain the plant extract composition.
Example 4: a method for preparing a plant extract composition having weight-losing and lipid-lowering activities, which is different from example 1, in that the concentration of the organic solvent aqueous solution is 40%.
Example 5: a method for preparing a plant extract composition having weight-losing and lipid-lowering activities, which is different from example 1, in that the concentration of the organic solvent aqueous solution is 95%.
Example 6: a method for preparing a plant extract composition having weight-losing and lipid-lowering activities, which is different from example 1 in that an aqueous solution of an organic solvent is replaced with an equal amount of water.
Comparative example
Comparative example 1: a preparation method of a plant extract composition with weight-losing and lipid-lowering activities is different from that of the embodiment 1 in that Pu' er tea is not added.
Comparative example 2: a method for preparing a plant extract composition with weight-losing and lipid-lowering activities, which is different from the method in example 1, is characterized in that grape skin is not added.
Comparative example 3: a preparation method of a plant extract composition with weight-losing and lipid-lowering activities is different from that of example 1 in that Pu' er tea and peony flowers are not added.
Comparative example 4: a preparation method of a plant extract composition with weight-losing and lipid-lowering activities is different from that of example 1 in that Pu' er tea and apple peel are not added.
Comparative example 5: a method for preparing plant extract composition with weight reducing and blood lipid reducing effects comprises defatting.
Comparative example 6: a method for preparing plant extract composition with weight reducing and blood lipid reducing effects comprises extracting in step S2 without ultrasound.
Comparative example 7: a method for preparing a plant extract composition with weight reducing and blood lipid reducing effects comprises in step S2, ultrasonic extracting at 80 deg.C.
Comparative example 8: cerulenin, available from Sigma-Aldrich, catalog No. 219557.
Comparative example 9: EGCG (Epigallocatachhin gate), available from Sigma-Aldrich under catalog number 93894.
Comparative example 10: resveratrol, available from Sigma-Aldrich, catalog number 34092.
Performance test
Firstly, detecting the inhibition activity of fatty acid synthase:
10mg of the product prepared in example or comparative example were dissolved in 200mL of DMSO as a test sample solution and then tested in a conventional in vitro assay for the enzymatic activity in tumor cells, i.e., after disruption of the tumor cells, the assay was carried out using standard assays using acetyl coenzyme A and malonyl coenzyme A, NADPH as substrates (TianW.X.et al, 1985.J.biol.chem, 260(20): 11375-11387; Li P, et al.,2014.Mol Cancer 13: 138).
The degree of inhibition of Fatty Acid Synthase (FAS) activity was expressed as IC50 (half inhibitory concentration) and was calculated as: determining the fatty acid synthase activity value of the substrate by using only the corresponding extraction solvent as a control, and using A as the value0Expressed and set to 100%. Adding fatty acid synthase inhibitor extract into substrate as experimental group, and measuring the value of fatty acid synthase activity as A0At 50%, the concentration of the inhibitor was the IC50 value, and the test results are reported in table 1.
TABLE 1 half inhibitory concentration assay results for fatty acid synthase inhibitory Activity
As can be seen from the data in Table 1, the semi-inhibitory concentration of fatty acid synthase by the plant extract compositions prepared in examples 1 to 3 was low, indicating that the plant extract compositions prepared in examples 1 to 3 of the present application have a strong inhibitory effect on the activity of fatty acid synthase and that the required effective dose was low.
In example 4, 40% by volume of an aqueous organic solvent solution and in example 5, 95% by volume of an aqueous organic solvent solution were used, and as compared with the results of the tests in examples 1 to 3, it was found that the fatty acid synthase inhibitory activity of examples 4 and 5 was decreased.
In example 6, water is used instead of the aqueous solution of the organic solution, and no organic solvent is added, so that the test result is greatly influenced.
Compared with the example 1, the half-inhibition concentration of the product prepared in the comparative example 1 on the fatty acid synthase is obviously increased, and compared with the test result in the comparative example 1, the test result of the comparative example 3 is larger than that in the comparative example 1, which shows that the Pu' er tea and the peony flower have better synergistic inhibition effect.
In comparative example 2, no grape skin was added, and the test results in comparative example 2 were larger than those in example 1, indicating that the fatty acid synthase inhibitory activity of the product prepared in comparative example 2 was decreased.
Compared with the comparative example 1, the test effect of the comparative example 4 is increased and the fatty acid synthase inhibiting activity is reduced because the Pu' er tea and the apple peel are not added in the comparative example 4.
Comparative example 5 compared to example 1, the non-defatted plant extract was not defatted and the fatty acids and other fat soluble components of the plant extract were not removed, resulting in a plant extract composition with more impurities, poor purity and reduced inhibitory activity against fatty acid synthase.
In comparative example 6, no sonication was performed while the extraction was performed, and as can be seen from the data in table 1, the fatty acid synthase inhibitory activity of the plant extract composition obtained without sonication was decreased.
Comparative example 7 compared with example 1, the temperature increased to 80 ℃ during the ultrasonic extraction, and the obtained plant extract composition had decreased inhibitory activity against fatty acid synthase, indicating that too high a temperature during the ultrasonic extraction is likely to cause destruction of the active ingredients of the plant extract composition.
In comparative example 8, cerulenin, a widely used natural Fatty Acid Synthase (FAS) inhibitor produced by the fungus Cephalosporium caeruleuus, was used, and its half inhibitory concentration against fatty acid synthase was increased to 22.5 μ g/mL, indicating that its inhibitory effect on the activity of fatty acid synthase was inferior to that of the plant extract composition prepared in the present application.
In comparative example 9, EGCG, epigallocatechin gallate, which is the main component of green tea polyphenol, is used, and catechin monomers separated from tea leaves have a half inhibitory concentration of 25.1. mu.g/mL against fatty acid synthase, and the inhibitory effect is inferior to that of the present application.
In comparative example 10, resveratrol, which is present in various plants, particularly grape skin, in a large amount, had a half inhibitory concentration of 11.6 μ g/mL against fatty acid synthase, and had inferior inhibitory activity to that of the present application.
It can be seen that the plant extract composition prepared herein has a significant inhibitory effect on the activity of fatty acid synthase, is more potent than the known fatty acid synthase inhibitors cerulenin, EGCG and resveratrol, and requires a very low effective dose.
Secondly, the weight-losing and food-inhibiting effects of the plant extract composition are as follows:
1. the weight-losing effect is as follows: 85 healthy subjects having a weight of (61 + -1) kg were selected, and divided into 17 groups on average, 5 persons in each group were measured for the weight of 5 persons in each group, and the average was found, 16 groups of subjects were orally administered with 500 mg each 3 times per day corresponding to the products prepared in examples 1 to 6 and comparative examples 1 to 10, group 17 was used as a control group, subjects were administered with fenfluramine as an antiobesity agent for 10mg per day, the weight of 10 persons in each group was measured after 16 days, the average was found, and the change in weight and the percentage (%) of weight loss were calculated, and the results of the measurement were recorded in Table 2.
2. Food inhibition effect: 170 mice are selected and randomly divided into 17 groups, 10 mice are selected, the 1 st group to the 13 th group are correspondingly fed with the products prepared in the examples 1 to 6 and the comparative examples 1 to 7, the 14 th group to the 16 th group are correspondingly injected with the products in the comparative examples 5 to 7, 0.2mL is injected every day, the 17 th group is used as a control group, double distilled water is fed, the feeding method comprises the steps of inserting a round head perfusion apparatus into the esophagus of the mice, perfusing the esophagus of the mice for 1 time every day, 0.2mL is injected every day, feeding is carried out at 10 to 11 am, the feed and drinking water are taken independently, the test time is 20d, the food intake of the mice is measured every 2 days, the average value of 10 measurement results is taken, and the measurement results are recorded in the table 2.
TABLE 2 weight loss and appetite control effects of plant extract compositions
After the plant extract composition prepared in the examples 1 to 3 is eaten by a subject daily, the weight of the subject is reduced by 2.25 to 3.1kg after 16 days, the weight reduction percentage is 3.69 to 5.17 percent, and the weight reduction effect is better, and when a food inhibition test is carried out on the subject by using a mouse, the food intake of the mouse is less within 20 days, so that the plant extract composition prepared in the examples 1 to 3 has good weight reduction and food inhibition effects, the weight reduction effect of the plant extract can be related to the inhibition of fatty acids and beauty, the plant extract prepared in the examples 1 to 3 is orally taken, and the three recognized fatty acid synthase inhibitors in the comparative examples 8 to 10 are all injected, so that the plant extract composition prepared in the application has wider application prospect.
Examples 4 and 5 used 40% and 95% by volume of aqueous organic solvent solutions, respectively, as compared to example 1, but the products produced in examples 4 and 5 had less weight loss and food inhibition than example 1.
Example 6 Using water as the extraction solvent, the resulting plant extract composition had a reduced weight loss value and increased food intake in mice compared to example 1.
Compared with the example 1, the weight reduction and food inhibition effects of the comparative example 1 and the comparative example 3 are poor because the Pu 'er tea is not added in the comparative example 1 and the Pu' er tea and the peony flowers are not added in the comparative example 3.
The grape skin was not added in comparative example 2, and the test results of comparative example 2 show that the product obtained in comparative example 2 has less effect on weight loss and food inhibition than example 1.
The Pu' er tea and the apple peel are not added in the comparative example 4, and detection shows that the weight of a subject eating the product of the comparative example 4 is not obviously reduced, the food intake of a mouse is higher, and the effects of weight loss and food inhibition are not good.
In comparative example 5, where no defatting was performed, it is shown in table 2 that the percent weight loss of the subjects was small compared to example 1, while the food intake of the mice was large, indicating that defatting increased the weight loss and food suppression effect of the plant extract plants.
In comparative example 6, the plant extract composition prepared without using ultrasound when extracted with an organic solvent had less weight loss and food suppression effects on the subjects than in examples 1 to 3.
Comparative example 7 the temperature was 80 ℃ at the time of sonication, as compared to example 1, and it is shown in table 2 that the weight loss value of the subject was decreased, the food intake of the mouse was increased, and the inhibitory and weight-loss effects were reduced.
The subjects in the control group received the anti-obesity drug, and after 16 days, the weight was reduced by only 0.65kg, while the mice receiving the double-distilled water through the stomach had a larger food intake than in examples 1-3, indicating that the plant extract composition prepared in this application had excellent weight loss and food inhibition effects.
Thirdly, detecting the effect of the plant extract composition on inhibiting tumor cells:
culturing three cancer cell strains of breast cancer MCF-7 (Shanghai Orlo Biotechnology Co., Ltd.), prostate cancer PC-3M 1E8 (Shanghai Jinma Biotechnology Co., Ltd.) and liver cancer BEL-7402 (Shanghai Minghai Haori Biotechnology Co., Ltd.) in RPMI-1640 culture solution containing 10% calf serum, penicillin and streptomycin, subculturing at 37 deg.C by attaching single layer to wall in an incubator containing 5% carbon dioxide, collecting three tumor cells in logarithmic growth phase, inoculating into 96-well plate, inoculating 2 × 10 tumor cells into each well4After 24 hours of adherent culture, the culture medium was changed to a culture medium containing the products prepared in examples or comparative examples, and 20. mu.L of tetramethyl coupler was addedAzole (5 mg/mL), continuing the culture for 4 hours, removing the supernatant, adding 200 μ L of dimethyl sulfoxide to each well (adding the same amount of culture solution and dimethyl sulfoxide to the control group), horizontally shaking for 10min, measuring the absorbance value (a) by an enzyme-linked immunosorbent assay, wherein the wavelength is 570nm, calculating the inhibition effect of the extract on the growth of cancer cells according to the following formula, the inhibition rate (%) of the growth of cancer cells is (%) = (1-experimental group a 570/control group a 570) × 100%, the concentration of the culture solution containing each example or comparative example is divided into 3 groups, which are 5 μ g/mL, 20 μ g/mL and 40 μ g/mL respectively, each concentration is provided with 3 multiple wells, averaging the test results, and recording the test results in table 3.
TABLE 3 inhibition of cancer cell growth by plant extract compositions
The plant extract compositions prepared in examples 1 to 3 have an increasing inhibition rate of cancer cell growth with increasing concentration, which shows that the plant extract compositions prepared in the present application have a significant effect of inhibiting cancer cell growth.
In example 4, 40% by volume of organic solvent aqueous solution is used, in example 5, 95% by volume of organic solvent aqueous solution is used, and the inhibition rate of the plant extract composition extracted in example 4 and example 5 on three tumor cells is reduced compared with that of example 1, and the effect of inhibiting the growth of cancer cells is reduced.
In example 6, water was used instead of the aqueous organic solvent solution, and the extracted plant extract composition had a lower inhibitory effect on cancer cells than in example 1 using water as the extractant.
The Puer tea is not added in the comparative example 1, the Puer tea and the peony flower are not added in the comparative example 3, the inhibition rate of the product prepared in the comparative example 1 on cancer cells is reduced, the inhibition rate of the comparative example 3 on the cancer cells is smaller than that of the comparative example 1, and the Puer tea and the peony flower can synergistically reduce the number of the cancer cells and inhibit the growth of the cancer cells.
In comparative example 2, grape skin was not added, and the inhibitory effect on liver cancer cells, prostate cancer cells, and breast cancer cells was reduced as compared with example 1.
Compared with the comparative example 1, the Puer tea and the apple peel are not added in the comparative example 4, so that the inhibition rate of the Puer tea and the apple peel on three cancer cells is reduced, and the Puer tea and the apple peel can achieve the effect of synergistically inhibiting the growth of the cancer cells.
Comparative example 5 compared with example 1, the degreasing step was not performed, and it can be seen from comparison of the data in table 3 that the obtained plant extract composition has a lower cancer cell inhibitory effect than that of example 1.
In comparative example 6, no ultrasound was performed during the extraction, and the product prepared in comparative example 6 had a reduced inhibitory effect on liver cancer cells, prostate cancer cells, and breast cancer cells, as compared to example 1.
In comparison with example 1, in comparative example 7, the temperature was increased to 80 ℃ during the ultrasonic treatment, and table 3 shows that the inhibition rate of comparative example 7 on three kinds of cancer cells is less than that of example 1, which indicates that the increase of the ultrasonic temperature affects the inhibition effect of the plant extract composition on the cancer cells.
In comparative examples 8 to 10, the currently accepted fatty acid synthase inhibitors were used, but the growth inhibitory effects of these three fatty acid synthase inhibitors on cancer cells were inferior to those of examples 1 to 3 of the present application.
The use of the plant extract composition prepared in examples 1-3 for the preparation of a medicament having weight loss or anticancer effects.
The use of the plant extract composition prepared in examples 1-3 for the preparation of a food product having weight loss or anticancer effects.
The use of the plant extract composition prepared in examples 1-3 for the preparation of a food additive having weight-loss or anticancer effects.
The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.
Claims (9)
1. A preparation method of a plant extract composition with weight-losing and lipid-lowering activities is characterized by comprising the following steps:
s1, mixing Pu' er tea, grape skin, apple skin and peony according to the weight ratio of 2-4:0.5-1.5:2-4:0.5-1, crushing and degreasing to obtain a raw material mixture;
s2, mixing the raw material mixture obtained in the step S1 with an organic solvent aqueous solution, carrying out ultrasonic extraction, and collecting an extracting solution to obtain a crude extracting solution;
s3, concentrating the crude extract obtained in the step S3 under reduced pressure until no organic solvent smell exists, further concentrating until the solvent is volatilized, and drying to obtain a target extract;
in the step S2, the concentration of the aqueous solution of organic solvent is 50-90% by volume.
2. The method of claim 1, wherein the ratio of the mixture of the raw materials to the organic solvent in step S2 is 1g (8-15) mL.
3. The method of claim 1, wherein the organic solvent in step S2 is selected from one of formic acid, acetic acid, methanol, ethanol, n-propanol, isopropanol, and acetone.
4. The method for preparing the plant extract composition with the activity of losing weight and reducing fat as claimed in claim 1, wherein the ultrasonic extraction in the step S2 is specifically performed by: ultrasonic extracting at room temperature for 1-3 times, each time for 10-30 min.
5. The method of claim 1, wherein the concentration under reduced pressure in step S3 is performed at 30-60 deg.C and 0.01-0.1 MPa.
6. The method for preparing the plant extract composition with the activity of losing weight and reducing fat as claimed in claim 1, wherein the defatting step S1 is as follows: adding pulverized Pu her tea, grape peel, apple peel and peony pollen into petroleum ether or cyclohexane, heating and refluxing in water bath at 30-60 deg.C for 10-30min for 2-3 times, and filtering to remove petroleum ether or cyclohexane.
7. A plant extract composition with weight-losing and lipid-lowering activities, which is prepared by the preparation method of the plant extract composition with weight-losing and lipid-lowering activities of any one of claims 1 to 6.
8. Use of the plant extract composition having weight-losing and lipid-lowering activities of claim 7 for the preparation of a medicament, food or food additive having weight-losing or anticancer effects.
9. A fatty acid synthase inhibitor comprising the plant extract composition having the activity of losing weight and reducing fat of claim 8.
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