CN107184658B

CN107184658B - Ligustrum lucidum extract, extraction method and application thereof in preparation of medicines for preventing and treating metabolic syndrome

Info

Publication number: CN107184658B
Application number: CN201710274380.1A
Authority: CN
Inventors: 康文艺; 崔丽丽; 马常阳; 邢梅梅
Original assignee: Henan University
Current assignee: Henan University
Priority date: 2017-04-25
Filing date: 2017-04-25
Publication date: 2020-10-27
Anticipated expiration: 2037-04-25
Also published as: CN107184658A

Abstract

The invention belongs to the field of medicines and/or health-care products, and particularly relates to a ligustrum lucidum flower extract, an extraction method and application thereof in preparing a medicine for preventing and treating metabolic syndrome. The privet flower extract is one or a mixture of more than two of a total extract of the privet flower, a 40% ethanol part and a 60% ethanol part which are extracted from the privet flower. The invention also discloses an extraction method of the glossy privet flower extract. The invention discovers through experiments that: the Ligustrum lucidum extract has the effects of preventing and treating metabolic syndrome, especially promoting fat cell sugar absorption, inhibiting preadipocyte differentiation and promoting fat metabolism of fat cells, and can be used for preparing medicines for preventing and treating diabetes and medicines for preventing and improving obesity.

Description

Ligustrum lucidum extract, extraction method and application thereof in preparation of medicines for preventing and treating metabolic syndrome

Technical Field

The invention belongs to the technical field of medicines and/or health-care products, and particularly relates to a ligustrum lucidum flower extract, an extraction method and application thereof in preparing a medicine for preventing and treating metabolic syndrome.

Background

Glossy privet (Ligustrum lucidumAit.) is a plant of the genus Ligustrum of the family Oleaceae. The fruit is used as medicine (glossy privet fruit)Ligustri Lucidi Fructus) Sweet, bitter and cool. It enters liver and kidney meridians. Has effects of nourishing liver and kidney, improving eyesight and blackening hair. Can be used for treating liver and kidney yin deficiency, giddiness, tinnitus, soreness of waist and knees, premature gray hair, dim eyesight, internal heat, diabetes, and hectic fever. Currently, the glossy privet fruit has a lot of researches and chemical compositionsThe main components include triterpenes, flavonoids, iridoids, phenethyl alcohol and the like. The pharmacological activities mainly comprise antioxidation, anti-inflammation, liver protection, anticancer, blood sugar reduction, immunoregulation and the like. In addition, there are reports that 19 compounds are identified by separating glossy privet bark, and compounds such as 7-hydroxycoumarin, benzoic acid, nicotinamide and the like are obtained in addition to the structural types of the compounds usually found in glossy privet. There is relatively little research on privet flowers. The volatile oil of Ligustrum lucidum ait flower mainly contains alcohol, aldehyde and ester. The chemical components of the privet flower mainly comprise flavonoids and sterols. Another research shows that the total flavonoids of glossy privet flower have strong capacity of eliminating DPPH free radicals and nitrite. At present, no study on the aspect of the metabolism syndrome of the Newcastle disease is seen.

Obesity is a chronic metabolic disease in which excessive fat accumulation and weight gain in a body are caused by various factors such as heredity and environment. Is closely related to type 2 diabetes and dyslipidemia. The intake of energy in excess of the energy expended can lead to fat accumulation. The increase in adipose tissue is caused by an increase in the number and volume of adipocytes. Abnormal proliferation and differentiation of adipose tissues causes accumulation of adipose tissues. Excessive accumulation of adipose tissues in a body can secrete a series of hormones and adipocytokines to interfere signal conduction of insulin in cells and initiate insulin resistance, wherein the insulin resistance refers to that the sensitivity of peripheral tissues (muscles and fat) in the body to insulin is reduced, so that the uptake of glucose by the peripheral tissues is reduced, the blood sugar is increased, and then the type 2 diabetes is induced. Also, insulin resistance extends throughout the entire process of type 2 diabetes. The main structure of adipose tissue is lipid droplets, with a content of triglycerides of 95%. Triglyceride is decomposed into free fatty acid and glycerol under the action of lipase, and the main function of the lipid decomposition is energy supply. Lipid droplets, also known as "energy converters", are subcellular structures associated with obesity, and intracellular lipid droplet enlargement can lead to increased adipocyte volume. The biological basis for obesity is: the increase in intracellular lipid droplets leads to an increase in cell volume and an increase in the number of new adipocytes. Therefore, the study of lipid droplet formation and triglyceride metabolism may be one of the means for solving the obesity problem. Adipocytes originate from mesodermal stem cells, which differentiate and develop through several stages: pluripotent stem cells, mesenchymal precursor cells, preadipocytes, mature adipocytes. The 3T3-L1 preadipocytes come from mouse embryonic fibroblasts, are cloned and amplified to form a preadipocyte system, can be directionally differentiated into mature adipocytes, fully show the characteristics of somatic cells, including morphological change, expression of various fat metabolic enzymes, extensive lipid accumulation and the like, and are ideal models for in vitro research of adipocytes. The increasing incidence of obesity and type 2 diabetes has become a global health problem. The development of natural products for preventing and treating obesity and type 2 diabetes is of great significance.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a ligustrum lucidum flower extract, an extraction method and application thereof in preparing a medicament for preventing and treating metabolic syndrome.

In order to achieve the purpose, the invention adopts the following technical scheme:

a Ligustrum lucidum flower extract is one or more of Ligustrum lucidum flower total extract, 40% ethanol part and 60% ethanol part.

The method for extracting the privet flower extract comprises the following steps: heating and reflux-extracting dry privet flower with petroleum ether for 1-3 times, reflux-extracting for 1-3h each time, performing solid-liquid separation after reflux-extracting is finished, volatilizing petroleum ether from the obtained residue, performing immersion-extraction for 2-4 times at room temperature by using 70 +/-5% ethanol, performing immersion-extraction for 2-4 days each time, combining the extracting solutions and concentrating to obtain a total extract of the privet flower; dissolving the total extract with ethanol, separating with D101 type macroporous resin, sequentially gradient eluting with water, 20% ethanol, 40% ethanol, 60% ethanol, and 95% ethanol, concentrating the eluate to obtain water part, 20% ethanol part, 40% ethanol part, 60% ethanol part, and 95% ethanol part. The privet flower extract is one or more of total extract, 40% ethanol part and 60% ethanol part of privet flower.

The application of the ligustrum lucidum flower extract in preparing the medicine for preventing and treating the metabolic syndrome.

The above application may specifically be: the application of the ligustrum lucidum flower extract in preparing the medicines for preventing and treating diabetes, in particular the application in preparing the medicines and/or health products for promoting the absorption of fat cell sugar, and further the application in preparing the medicines and/or health products for promoting the absorption of fat cell sugar under the insulin resistance state.

The above application may specifically be: the privet flower extract has potential weight-losing efficacy, in particular to the application in preparing medicines and/or health-care products for inhibiting the differentiation of preadipocytes, or the application in preparing medicines and/or health-care products for promoting the lipid metabolism of adipocytes.

The privet flower extract can also be compounded with conventional auxiliary materials in the field to prepare a compound preparation. The compound preparation can be prepared into tablets, granules, capsules, pills, injections and the like.

Compared with the prior art, the invention discovers through experiments that the ligustrum lucidum flower extract can be a novel compound for treating type 2 diabetes and obesity by inhibiting the differentiation of preadipocytes, promoting the lipid metabolism of adipocytes, promoting the sugar absorption of adipocytes and improving insulin resistance, namely the ligustrum lucidum flower extract has the potential effects of reducing blood sugar and improving insulin resistance. Therefore, the compound can be used for preparing medicines and/or health products for preventing and treating metabolic syndrome, in particular to medicines for preventing and improving diabetes and obesity.

Drawings

FIG. 1 shows the results of oil-red differentiation staining of the control group;

FIG. 2 shows the results of oil-red differentiation staining of the total extract of Ligustrum lucidum;

FIG. 3 shows the results of oil-red differentiation staining of the 40% ethanol part of glossy privet flower;

FIG. 4 shows the results of oil-red differentiation staining of the 60% ethanol part of glossy privet flower;

FIG. 5 shows the effect of Ligustrum lucidum extract on lipid metabolism by 3T3-L1 adipocytes;

FIG. 6 shows the sugar absorption effect of Ligustrum lucidum extract on 3T3-L1 adipocytes;

FIG. 7 is a graph showing that dexamethasone induces the formation of insulin resistance in 3T3-L1 mature adipocytes;

FIG. 8 shows the effect of Ligustrum lucidum extract on glucose absorption by 3T3-L1 adipocytes in an insulin resistant state.

Detailed Description

The technical solution of the present invention is further described in detail with reference to the following examples, but the scope of the present invention is not limited thereto.

In the following examples, the percentages of ethanol are by volume unless otherwise specified.

Example 1

A Ligustrum lucidum flower extract is one or more of Ligustrum lucidum flower total extract, 40% ethanol part extract and 60% ethanol part extract.

The method for extracting the privet flower extract specifically comprises the following steps:

1) adding petroleum ether into 475 g of dried privet flower until the privet flower is immersed, then heating and refluxing for extraction for 2 times, wherein the reflux extraction is performed for 2 hours each time, after the reflux extraction is finished, filtering, volatilizing the petroleum ether from obtained residues, performing immersion extraction for 3 times (the addition amount of 70% ethanol is proper for immersing the residues) at room temperature by using 70% ethanol for 3 days each time, combining extracting solutions and concentrating to obtain a total extract of the privet flower;

2) dissolving the glossy privet flower total extract with 50% ethanol, adsorbing and separating with D101 type macroporous resin, sequentially carrying out gradient elution with water, 20% ethanol, 40% ethanol, 60% ethanol and 95% ethanol, wherein each gradient elution has five column volumes, each column volume is 2000 mL, and concentrating the eluent to obtain a water part, a 20% ethanol part 7g, a 40% ethanol part 60g, a 60% ethanol part 24g and a 95% ethanol part 16 g.

Application test 1, influence of Ligustrum lucidum flower extract on 3T3-L1 preadipocyte activity

The 3T3-L1 preadipocytes are taken as a cell model, the ligustrum lucidum extract prepared in the example 1 is added in the process of inducing differentiation, and the degree of differentiation of the 3T3-L1 preadipocytes is identified by oil red O staining.

Instrument for measuring the position of a moving objectMaterials: the instrument comprises the following steps: preparative high performance liquid chromatography (Waters 2535Q); a rotary evaporator (EYELA, Tokyo physical and chemical instruments Co., Ltd.); LC-3000 high performance liquid preparative column chromatography (Beijing Innovation science and technology Co., Ltd.); am-400 superconducting nuclear magnetic resonance apparatus (Bruker); electronic balance (Mettler-Toledo, USA); TGL-16gR high speed table refrigerated centrifuge (Shanghai' an Tingning scientific Instrument plant); a double single-side purification workbench (model: SW-CJ-2FD type manufacturer: Suzhou purification Equipment Co., Ltd.); a carbon dioxide incubator (model: 3111, manufacturer: Thermo scientific); inverted microscope (model ckx31, olympus); a full-temperature shaking incubator (HZP type shanghai sperm macroexperimental facilities ltd); centrifugal precipitators (type 800 Shanghai surgical instruments factory); various sizes of pipette (transferpette); digital camera (canon EOS 450D) inverted microscope (Nikon ECLiPSE TS 100); GF 254 silica gel thin layer plates (tai huiyou silica gel development ltd); 40-80 mesh silica gel, 200-300 mesh silica gel and silica gel H (Nicoti Huiyou silica gel development Co., Ltd.); sephadex LH-20 (Pharmacia, Sweden); d101 type macroporous resin (tianjin maritime chemical limited); c-18 (Merk, Germany); the other reagents are analytically pure; glucose assay kit (shanghai nabobism pharmaceutical limited, 20161105147); glucose standards (Shanghai Rongsheng biopharmaceutical Co., Ltd., lot number: 20161001); mouse free fatty acid assay kit (Nanjing Sen Beiga Biotech limited, lot number: 201609); fetal bovine serum (Zhejiang space navigation Biotechnology GmbH, lot number: 20160923); DMEM high-glucose medium (Beijing Soilebao Tech., batch No.: F08HV 090); penicillin streptomycin mixture (solarbio, lot # 20161029); pancreatin cell digestive juice (Biyunyan, No. C0210); dimethyl sulfoxide (sigmaSHBC 3313V); insulin (sigma); thiazole blue (Shanghai Hualan science and technology Co., Ltd.); oil red O (sigmaSLBP 5248V); IBMX (sigma, BCBH 1214V); DeX (sigma BCBM 4557V). Rosiglitazone tablets (medhenri pharmaceutical limited); lovastatin capsules (Yangziang pharmaceutical industry group Co., Ltd., 20160109); cell lines: 3T3-L1 mouse embryo fibroblastsCells were purchased from a cell bank of the Chinese academy of sciences. Glossy privet flower is collected in Huaxi district wetland park of Guiyang city of Guizhou province in 2015 6 months, and is identified as glossy privet by professor Zhang-Qian-Jun of Guizhou university (glossy privet)Ligustrum lucidumAir.). Specimens are presented at the institute of traditional Chinese medicine, university of Henan.

The experimental method comprises the following steps:

cell culture and cryopreservation:

3T3-L1 preadipocytes were cultured in DMEM high-glucose medium (containing penicillin: 10U/mL, streptomycin 10. mu.g/mL) containing 15% fetal bovine serum at 37 ℃ with 5% CO₂Culturing in a carbon dioxide incubator with saturated humidity, changing the culture solution once 2 d, removing the culture solution in the culture bottle when the cells are fused to 80%, adding 1mL of 0.25% trypsin solution, acting for about 1min, adding 2 times of culture solution to stop digestion, and blowing adherent cells into the culture solution by using a blow-beating tube. The cell suspension was transferred to a sterile centrifuge tube, centrifuged at 1000 rpm for 5 min and the supernatant discarded. Cell passage: cell pellets collected from 1 flask were blown off with 1mL of fresh medium, as 1: and (5) carrying out passage at a ratio of 2. Freezing and storing cells: the cell sediment collected from 2 culture bottles was blown up with 1.2 mL of a cryopreservation solution (fetal bovine serum containing 10% DMSO), transferred into a cryopreservation tube, placed into a programmed cooling box, then placed into a refrigerator at-80 ℃ overnight, and the cryopreservation tube was taken out and placed into a liquid nitrogen tank for storage.

And (3) detecting the cytotoxic activity:

3T3-L1 preadipocytes from the logarithmic growth phase were collected and the cell suspension concentration was adjusted to 2X 10 per well⁵The density of (2) was inoculated in a 96-well plate, and 100. mu.L of cell suspension was added per well. The cells were incubated at 37 ℃ with 5% CO₂And culturing in a carbon dioxide incubator with saturated humidity for 24 h until the cells are fused to 60% -70%, adding the medicine with concentration gradient, arranging a normal control group, a solvent control group (0.1% DMSO) and an extract administration group on each plate, arranging 6 multiple wells, continuously culturing for 48 h, adding 10 mu LMTT solution (5 g/L) into each well, and continuously culturing for 4 h. Carefully remove the culture medium in the wells, add 100. mu.L DMSO per well, shake the wells in a shake incubator (100 r/min) for 10 min to dissolve the crystals sufficiently. The absorbance value (OD value) of each well was measured at a wavelength of 570 nm in a microplate reader.

Table 1: glossy privet flower total extract and 60% ethanolEffect of site on the cell Activity of 3T3-L1 preadipocytes (n=6）

As can be seen from Table 1, the difference between the absorbance values of the normal control group and the vehicle control group is not statistically significant, indicating that the solvent does not affect the cell viability. Compared with the absorbance values of a normal control group, the differences of the total extract of the privet flower and the 60% ethanol part dosage group have no statistical significance, which indicates that neither the total extract of the privet flower nor the 60% ethanol part dosage group has cytotoxic activity, namely that the total extract of the privet flower and the 60% ethanol part can be set as the administration dosage below the dosage range of 300 mu g/mL.

TABLE 2 Effect of 40% ethanol fraction of Ligustrum lucidum on the Activity of 3T3-L1 preadipocytes (n=6）

As shown in Table 2, the difference between the absorbance values of the normal control group and the vehicle control group is not statistically significant, which indicates that the solvent does not affect the cell viability. Compared with the absorbance values of the normal control group, the difference of the 40% ethanol part dose group has no statistical significance, and the result shows that all the 40% ethanol part dose groups have no cytotoxic activity. That is, the dose of the 40% ethanol fraction can be set to a dose of 300. mu.g/mL or less.

Application test 2, influence of Ligustrum lucidum flower extract on differentiation of 3T3-L1 preadipocytes

Subculturing 3T3-L1 preadipocytes of logarithmic growth phase at a ratio of 1.5 × 10⁵The cells were inoculated in a 96-well plate, after complete fusion, the cells were subjected to contact inhibition for 48 hours, and then a DMEM high-glucose medium (containing 15% fetal bovine serum) containing a differentiation inducer A (0.5 mM IBMX, 1. mu.M DeX, 10. mu.g/mL Insulin) was replaced while adding drugs (blank group, model group, lovastatin group (50. mu.M), administration group). Changing into DMEM high-sugar culture solution (containing 15% fetal calf serum) containing an induction differentiation agent B (containing 10 mug/mL of Insulin) after 3 days, changing into conventional culture solution after 2 days,after that, the culture medium was changed every other day, cultured to 12 days, carefully aspirated, washed with ice-cold PBS 2 times, air-dried, fixed with 50. mu.L of 4% paraformaldehyde per well for 40 min, washed with PBS 2 times again, air-dried, stained with oil red O for 30 min, washed with PBS 2 times, washed with ultrapure water 2 times, and photographed with a digital camera under an inverted microscope (100X). Add 100. mu.L of isopropanol to each well and after 40 min measure the absorbance at 495 nm using a microplate reader. The results are as follows.

Data processing: results are expressed as mean ± SD values, and data statistics were compared for significant differences using SPSS19.0 software One-Way ANOVA (One-Way ANOVA).

Table 3: influence of Ligustrum lucidum flower Total extract on differentiation of 3T3-L1 preadipocytes (n=6）

As shown in Table 3 and FIGS. 1 and 2, the oil red staining level was significantly reduced in the lovastatin group and the glossy privet flower total extract at 300. mu.g/mL and 100. mu.g/mL, as compared with the model group (see the results of the present invention) ((P< 0.05), the level of oil red staining was significantly reduced at 33, 11. mu.g/mL of Ligustrum lucidum (L.) W.T. ((L.) W.P< 0.01). The lovastatin differentiation inhibition rate is: 35.27 percent, the differentiation inhibition rate of the total extract of the glossy privet flower is respectively as follows: 18.39%, 21.80%, 32.68%, 31.62%. The lovastatin and the glossy privet flower total extract can inhibit the differentiation of preadipocytes, so that the lovastatin and the glossy privet flower total extract have potential for preventing and improving obesity.

TABLE 4 influence of 40% ethanol fraction of Ligustrum lucidum on differentiation of 3T3-L1 progenitors (n=6）

As can be seen from table 4 and fig. 1 and 3, the oil red staining levels of all the dose groups of the 40% ethanol part of the ligustrum lucidum were not changed, indicating that the 40% ethanol part of the ligustrum lucidum could not inhibit the differentiation of 3T3-L1 preadipocytes.

Table 5: influence of 60% ethanol fraction of Ligustrum lucidum on differentiation of 3T3-L1 preadipocytes (n=6）

From table 5, fig. 1 and 4, it can be seen that the oil red staining level of the 60% ethanol part of glossy privet flower in the 300 μ g/mL dose group was significantly reduced (P<0.001), which indicates that the glossy privet flower 60% ethanol portion 300 μ g/mL dose group can inhibit the differentiation of 3T3-L1 preadipocytes, wherein the glossy privet flower 60% ethanol portion 300 μ g/mL dose group has a differentiation inhibition rate of 24.24%. The oil red staining levels of the glossy privet flower 60

% ethanol parts

100, 33 and 11 mug/mL dose groups were not changed, which indicates that the glossy privet flower 60

% ethanol parts

100, 33 and 11 mug/mL dose groups could not inhibit the differentiation of 3T3-L1 preadipocytes.

Application test 3 influence of Ligustrum lucidum flower extract on lipid metabolism of 3T3-L1 fat cells

Subculturing 3T3-L1 preadipocytes of logarithmic growth phase at a ratio of 1.5 × 10⁵The cells are inoculated into a 96-well plate, after the cells are completely fused, the cells are contacted and inhibited for 48 hours, then DMEM high-sugar culture solution (containing 15% fetal calf serum) containing a differentiation inducer A (0.5 mM IBMX, 1 mu M DeX and 10 mu g/mL Insulin) is replaced, DMEM high-sugar culture solution (containing 15% fetal calf serum) containing an induction differentiation agent B (10 mu g/mLInsulin) is replaced after 3 days, DMEM high-sugar culture solution containing 15% fetal calf serum is replaced after 2 days, the culture solution is replaced every other days, and the cells are cultured to 12 days to be induced and differentiated into mature adipocytes. Mature adipocytes were grouped and dosed (blank group, rosiglitazone group (9.52. mu.M), dosed group), cultured for another 48 h, cell culture supernatant was collected in a sterile tube, centrifuged at 2600 g for 20 min, and the supernatant was carefully collected. The content of free fatty acid was determined using a mouse free fatty acid assay kit. The results are as follows.

FIG. 5 shows the effect of Ligustrum lucidum extract on lipid metabolism of 3T3-L1 adipocytes. As can be seen from FIG. 5, the content of free fatty acids in the Total Extract (TE) of Ligustrum lucidum ait flower of 100. mu.g/mL and the dose group of 40% ethanol fraction (40% EF) of 33. mu.g/mL are higher than those in the blank group, the difference has statistical significance (P < 0.05), the content of free fatty acids in the dose group of 40% ethanol fraction of Ligustrum lucidum ait flower of 11. mu.g/mL is higher than those in the blank group, the difference has statistical significance (P < 0.05), and the content of free fatty acids in the dose group of 60% ethanol fraction (60% EF) of 11. mu.g/mL is higher than those in the blank group, which indicates that the total extract of Ligustrum lucidum ait flower of 100. mu.g/mL, the ethanol fraction of 33, 11. mu.g/mL and the ethanol fraction of 40% and 11. mu.g/mL can promote the decomposition and metabolism of triglyceride in fat cells into free fatty acids, it is shown that it can be used for promoting fat metabolism of fat cells and has potential for preventing and improving obesity.

Application test 4 influence of Ligustrum lucidum flower extract on sugar absorption of 3T3-L1 mature fat cells

The preadipocytes were induced to differentiate into mature adipocytes, and the experimental procedure was the same as in application test 3. Adding medicine (blank group, rosiglitazone group (9.52 μ M), administration group), culturing for 48 hr, collecting culture supernatant, and detecting glucose content in the culture solution according to glucose determination kit.

FIG. 6 shows the sugar absorption effect of the total extract, the 40% ethanol part and the 60% ethanol part of the privet flower on 3T3-L1 adipocytes. As shown in FIG. 6, the differences between rosiglitazone, 300, 11 μ g/mL total extract of glossy privet flower, 33, 11 μ g/mL 40% ethanol fraction and 100 μ g/mL60% ethanol fraction were statistically significant (the difference is below that of the blank group) ((P<0.05). The glucose content of the total extract of 33 mug/mL and 60 percent ethanol of 33 mug/mL and 11 mug/mL is lower than that of the blank group, and the difference has statistical significance (P)<0.01), which shows that rosiglitazone, 300, 33 and 11 mug/mL total extract, 33 and 11 mug/mL 40% ethanol part, 100, 33, 11 mug/mL 60% ethanol part can promote the uptake of peripheral glucose by fat cells, and the glucose content in the culture medium is reduced to a greater extent than that in a blank group, thus the potential of preventing and treating diabetes mellitus is realized.

Application test 5, Effect of Ligustrum lucidum extract on sugar absorption of 3T3-L1 mature adipocytes in insulin resistant state

The preadipocytes were induced to differentiate into mature adipocytes, and the experimental procedure was the same as in application test 3. Mature adipocytes were divided into blank and induced resistance groups. And (3) giving 1 mu M dexamethasone to the induced resistance group for acting for 3 d, taking the supernatant to determine the glucose content, and identifying the success of model building of the insulin resistance model. The insulin resistance group was dosed (blank group, rosiglitazone group (20. mu.M), and the supernatant was taken to determine the sugar value.

FIG. 7 is a graph showing that dexamethasone induces the formation of insulin resistance in 3T3-L1 mature adipocytes. As can be seen from FIG. 7, the glucose content in the resistant group was higher than that in the model group, and the difference was statistically significant (P < 0.001). The results show that the uptake of peripheral glucose by resistant adipocytes is reduced, and the model of insulin resistance model is successfully modeled.

FIG. 8 shows the effect of the total extract of Ligustrum lucidum, the 40% ethanol fraction, and the 60% ethanol fraction on the glucose absorption of 3T3-L1 adipocytes in the insulin resistant state. As can be seen from FIG. 8, the glucose content of the total extract of rosiglitazone, 33, 11. mu.g/mL of Ligustrum lucidum ait and the glucose content of the 60% ethanol fraction of 33, 11. mu.g/mL were lower than that of the blank group, and the difference was statistically significant (P < 0.05). 300. The glucose content of the total extract of 100 mu g/mL is lower than that of the blank group, and the difference has statistical significance (P is less than 0.01). The difference between the 40% ethanol fraction and the blank was not statistically significant. All the dosage groups of the glossy privet flower total extract and the 60% ethanol part with the concentration of 33 mu g/mL and 11 mu g/mL can promote the fat cells to take the peripheral glucose under the insulin resistance state, and the preparation method can be used for preventing and treating diabetes.

And (4) conclusion: the privet flower extract can inhibit the differentiation of preadipocytes, promote the lipid metabolism of adipocytes, promote the sugar absorption of adipocytes and improve insulin resistance, so that the privet flower extract can be a novel compound for treating type 2 diabetes and obesity, namely the privet flower extract has potential effects of reducing blood sugar and improving insulin resistance. Therefore, the compound can be used for preparing medicines and/or health products for preventing and treating metabolic syndrome, in particular to medicines for preventing and improving diabetes and obesity.

Claims

1. A privet flower extract for preventing and treating metabolic syndrome is characterized in that the privet flower extract is one or a mixture of more than two of a total extract of the privet flower, a 40% ethanol part and a 60% ethanol part;

the privet flower extract is obtained by the following steps: collecting dried glossy privet flowerLigustrum lucidumAit, with petroleum etherHeating and reflux-extracting for 1-3 times, each time for 1-3h, performing solid-liquid separation after reflux extraction is finished, volatilizing petroleum ether from the obtained residue, soaking and extracting for 2-4 times at room temperature by using 70 + -5% ethanol, each time for 2-4 days, combining the extracting solutions and concentrating to obtain the total extract of the privet flower; dissolving the total extract with ethanol, separating with D101 type macroporous resin, sequentially gradient-eluting with water, 20% ethanol, 40% ethanol, 60% ethanol, and 95% ethanol, concentrating the eluate, and collecting 40% ethanol part and 60% ethanol part.

2. The method for extracting the privet flower extract as claimed in claim 1, wherein the method comprises the steps of taking dry privet flower, heating and refluxing the dry privet flower with petroleum ether for 1-3 times, wherein the reflux extraction is performed for 1-3 hours each time, performing solid-liquid separation after the reflux extraction is finished, volatilizing the petroleum ether from the obtained residue, performing immersion extraction with 70 +/-5% ethanol at room temperature for 2-4 times, performing immersion extraction for 2-4 days each time, combining the extracting solutions and concentrating to obtain a total extract of the privet flower; dissolving the total extract with ethanol, separating with D101 type macroporous resin, sequentially gradient-eluting with water, 20% ethanol, 40% ethanol, 60% ethanol, and 95% ethanol, concentrating the eluate, and collecting 40% ethanol part and 60% ethanol part.

3. Use of the extract of ligustrum lucidum ait of claim 1 for the preparation of a medicament for the prevention and treatment of metabolic syndrome.

4. The use according to claim 3, wherein the Ligustrum lucidum extract is used for the preparation of a medicament for the prevention and treatment of diabetes.

5. The use according to claim 4, wherein the extract of Ligustrum lucidum ait is used in the preparation of a medicament for promoting the absorption of sugar by adipocytes.

6. The use of claim 3, wherein the extract of Ligustrum lucidum ait is used for the preparation of a medicament for preventing and improving obesity.

7. The use of claim 6, wherein the extract of Ligustrum lucidum ait is a total extract of Ligustrum lucidum ait and/or a 60% ethanol fraction of Ligustrum lucidum ait in the manufacture of a medicament for inhibiting differentiation of preadipocytes.

8. The use according to claim 6, wherein the extract of Ligustrum lucidum ait is used for the preparation of a medicament for promoting lipid metabolism in adipocytes.