Songorica soongarica flower extract and application thereof in preparation of weight-losing products

Abstract

The invention provides a salix soongarica flower extract and application thereof in preparation of weight-losing products, and belongs to the technical field of extraction of effective components of Chinese herbal medicines. According to the present invention, the product extracted from the soongarica soongarica flower through 70% alcohol and subjected to rotary evaporation concentration has effects of high fat diet induced animal weight reducing, liver lipid accumulation improving and liver inflammation improving, can be used for preparing the fat reducing product, and has good market application prospects.

Description

Technical Field

[0001] The invention belongs to the technical field of extraction of effective components of Chinese herbal medicines, and in particular relates to a Junggar willow flower extract and application thereof in preparing a weight loss product. Background Art

[0002] Obesity is a complex disease caused by the interaction of genetics and the environment, and is also a risk factor for many diseases, such as cardiovascular disease, non-alcoholic liver disease, diabetes and cancer. Currently, commonly used weight loss products are generally ineffective and have significant side effects, so safe, efficient and natural weight loss products have become a hot topic of research. Natural plant extracts can affect obese bodies, such as reducing weight, reducing body fat, improving insulin abnormalities, and improving inflammation.

[0003] Willow is a woody plant of the genus Salix in the family Salicaceae. It has many species and is not only one of the important plants for landscaping, but also its roots, bark, twigs and leaves have medicinal value. For example, the patent document with application number CN 106236828 A discloses the application of a Cyclocarya paliurus leaf extract in traditional Chinese medicine. The Cyclocarya paliurus compound and the Cyclocarya paliurus leaf water extract can improve the Yin deficiency score of the rat model of Yin deficiency and internal heat, and have the effects of nourishing Yin and clearing heat; can regulate the disordered cellular immune function and improve the state of deficiency syndrome; has the effect of lowering blood sugar and improving the state of insulin resistance; for another example, the patent document with application number CN 102558254A discloses a preparation method for extracting salicin from willow bark or willow branches, the main uses of which are anti-inflammatory, analgesic, and antipyretic. Salicin has similar biological activities to aspirin (acetylsalicylic acid), and is non-irritating to the stomach and is not easy to cause gastric bleeding. Junggar willow (Salix songarica) has strong adaptability, dense crown and developed root system. It is an excellent tree species for maintaining water and soil in desert and semi-desert river areas. As a wild willow of the genus Salix in the Salicaceae family unique to Xinjiang, after virus infection or insect pests, the growth of young branches and buds of Junggar willow is stunted and chrysanthemum-like symptoms are formed, which are called Junggar willow flowers. It has great potential for medicinal value. Summary of the invention

[0004] In view of this, the object of the present invention is to provide an application of a willow flower extract in the preparation of a weight loss product.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention discloses a willow flower extract of Junggari, which is obtained by extracting willow flower powder with an organic solvent.

[0007] Preferably, the organic solvent is ethanol.

[0008] Further preferably, the ratio of the willow flower powder to ethanol is 1 g:15-25 mL.

[0009] The above-mentioned extract is preferably extracted at a temperature of 85-95°C.

[0010] Furthermore, after the extraction is completed, rotary evaporation is used for concentration.

[0011] Application of the above extract in preparing fat-reducing products.

[0012] Application of the above extract in the preparation of a drug for preventing obesity caused by a high-fat diet

[0013] The application of the extract in preparing products for regulating serum hormones.

[0014] Preferably, the serum hormones include leptin, insulin and neuropeptide Y.

[0015] Application of the extract in preparing liver protection products.

[0016] Beneficial effects:

[0017] The present invention provides an application of a Junggar willow flower extract, which uses ethanol to extract effective ingredients from the Junggar willow flower. Under the premise that there is no difference in energy intake, the extract has the effects of reducing obesity, regulating serum hormone levels, reducing liver weight, improving liver inflammation induced by a high-fat diet, and reducing lipid accumulation. The extract can be used to prepare fat-reducing functional products and has good market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 : Experimental results of Example 1, A is the picture of mice in each group after 8 weeks of intervention; B is the weight change of mice during the 8 weeks of intervention; C is the weekly energy intake of each mouse; D is the final weight of the mouse; E is the Lee's index; F is the weight gain.

[0019] Figure 2 : INS levels in the serum of mice of Example 1.

[0020] Figure 3 : LEP levels in the serum of mice in Example 1.

[0021] Figure 4 : NPY levels in the serum of mice of Example 1.

[0022] Figure 5 : Liver weight of the mice in Example 1.

[0023] Figure 6 : Per-WAT index of the mice in Example 1.

[0024] Figure 7 : Epi-WAT index of the mice in Example 1.

[0025] Figure 8 : BAT index of the mice in Example 1.

[0026] Fig. 9 : Pictures of the mouse liver sections after H&E staining in Example 1.

[0027] Fig.10 : Pictures of mouse liver sections after OPR staining in Example 1. DETAILED DESCRIPTION

[0028] The invention provides a Junggar willow flower extract, which is a willow flower extract obtained by extracting willow flower powder with an organic solvent.

[0029] Optionally, the willow flower powder is obtained by drying the willow flowers in the shade, grinding them into powder, and passing through a 30-100 mesh sieve, preferably through a 50 mesh sieve.

[0030] An optional organic solvent is ethanol. Preferably, the ethanol is 65-75% ethanol. More preferably, the concentration of the ethanol is 70%.

[0031] Optionally, the ratio of the willow flower powder to ethanol is 1:15-25 g / mL. Preferably, the ratio of the willow flower powder to ethanol is 1:20 g / mL.

[0032] The optional extraction methods of the above extracts include immersion method, percolation method, ultrasound-assisted extraction method, reflux method and constant temperature water bath method. Preferably, the constant temperature water bath method is used.

[0033] Optionally, the extraction temperature is 85-95°C. Preferably, the temperature is 90°C.

[0034] Optionally, the extraction time in a constant temperature water bath is 1 to 2 hours. Preferably, the extraction time is 1.5 hours.

[0035] When the extract is extracted, further concentration is required after the extraction is completed. Preferably, the concentration method includes one or two of freeze drying and rotary evaporation. More preferably, the concentration method adopts rotary evaporation. Most preferably, the thick paste extract obtained by rotary evaporation is concentrated.

[0036] Application of the above extract in preparing fat-reducing products.

[0037] Application of the above extract in the preparation of a drug for preventing obesity caused by a high-fat diet

[0038] The application of the extract in preparing products for regulating serum hormones.

[0039] Preferably, the serum hormones include leptin, insulin and neuropeptide Y.

[0040] Application of the extract in preparing liver protection products.

[0041] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0042] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0043] In the experiments of the examples listed below: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001

[0044] Example 1

[0045] A kind of Junggar willow flower extract

[0046] The flowers of Junggar willow were picked, dried in the shade, ground into powder, and passed through a 50-mesh sieve to obtain Junggar willow flower powder. The extract was 70% alcohol, with a solid-liquid ratio of 1g:20mL, and was extracted in an electric constant temperature water bath at 90°C for 1.5h, and concentrated by rotary evaporation to obtain Junggar willow flower extract (SSE) thick paste.

[0047] Example 2

[0048] The flowers of Junggar willow were picked, dried in the shade, ground into powder, and passed through a 50-mesh sieve to obtain Junggar willow flower powder. The extract was 65% alcohol, with a solid-liquid ratio of 1g:15mL, and was extracted in an electric constant temperature water bath at 85°C for 1h, and concentrated by rotary evaporation to obtain Junggar willow flower extract (SSE) thick paste.

[0049] Example 3

[0050] The flowers of Junggar willow were picked, dried in the shade, ground into powder, and passed through a 100-mesh sieve to obtain Junggar willow flower powder. The extract was 75% alcohol, with a solid-liquid ratio of 1g:25mL, and was extracted in an electric constant temperature water bath at 95°C for 1.5h, and concentrated by rotary evaporation to obtain Junggar willow flower extract (SSE) thick paste.

[0051] Example 4

[0052] The flowers of Junggar willow were picked, dried in the shade, ground into powder, and passed through a 30-mesh sieve to obtain Junggar willow flower powder. The extract was 70% alcohol, with a solid-liquid ratio of 1g:20mL, and was extracted in an electric constant temperature water bath at 90°C for 1h, and concentrated by rotary evaporation to obtain Junggar willow flower extract (SSE) thick paste.

[0053] Example 5

[0054] The flowers of Junggar willow were picked, dried in the shade, ground into powder, and passed through a 50-mesh sieve to obtain Junggar willow flower powder. The extract was 75% alcohol, with a solid-liquid ratio of 1g:20mL, and was extracted in an electric constant temperature water bath at 90°C for 2h, and concentrated by rotary evaporation to obtain Junggar willow flower extract (SSE) thick paste.

[0055] Example 6

[0056] The flowers of Junggar willow were picked, dried in the shade, ground into powder, and passed through a 100-mesh sieve to obtain Junggar willow flower powder. The extract was 65% alcohol, with a solid-liquid ratio of 1g:20mL, and was extracted in an electric constant temperature water bath at 90°C for 2h, and freeze-dried at -80°C to obtain Junggar willow flower extract powder.

[0057] Example 7

[0058] Fat-reducing effect of extract of Junggar willow flower

[0059] Fifty 7-week-old male C57BL / 6J mice (purchased from Henan Sikebes Biotechnology Co., Ltd.) were selected and kept under constant temperature and humidity conditions (22±2℃, humidity 55%±5%) with a 12-h light-dark cycle, and free access to food and water. After one week of acclimation, the mice were randomly divided into a normal diet group and a high-fat diet group (9 mice in the normal diet group and the rest in the high-fat diet group, normal diet: 3.85kcal / g calories, 20% protein, 70% carbohydrates, 10% fat; high-fat diet: 5.24kcal / g calories, 20.00% protein, 20.00% carbohydrates, 60.00% fat), and the average weight of mice in the high-fat diet group was 20% higher than the average weight of mice in the normal diet group as the modeling success standard.

[0060] After the modeling was successful, the mice were redivided into 6 groups with 6 mice in each group, namely normal diet (ND) group, high-fat diet (HFD) group, orlistat group, high-dose (H) group of SSE prepared in Example 1, medium-dose (M) group of SSE prepared in Example 1 and low-dose (L) group of SSE prepared in Example 1. Among them, the ND group was fed with ordinary feed (calories of 3.85 kcal / g, protein 20%, carbohydrates 70%, fat 10%), and the other 5 groups were fed with high-fat feed (calories of 5.24 kcal / g, protein 20.00%, carbohydrates 20.00%, fat 60.00%). The ND group and the HFD group were gavaged with an equal amount of normal saline, the Orlistat group was gavaged with Orlistat solution at a dose of 50 mg / kg, and the H group was gavaged with 5.88 g·kg -1 ·d -1 The M group was gavaged with 2.94 g kg -1 ·d -1 SSE solution, L group was gavaged 1.47 g kg -1 ·d -1 The SSE solution was intragastrically administered for 8 weeks.

[0061] The weight of mice was measured and recorded every week, and the body length of each mouse was measured before and after the SSE intervention experiment. The weight, energy intake and Lee's index of mice were calculated every week. At the end of 8 weeks, the mice were dissected and the fat index was calculated.

[0062] Energy intake: Each mouse was provided with an average of 3 g of feed per day. The food intake was counted once a week, and the weight changes of each mouse were measured and recorded every week.

[0063] Fat index = (epididymal fat + perirenal fat + brown fat) g / body weight g × 100%.

[0064] Lee's index = 1 / 3 body weight (g) × 10 3 / Body length (cm).

[0065] Experimental results:

[0066] from Figure 1 It can be seen that after 8 weeks of gavage, the final body weight, weight gain, Lee's index and weekly energy intake of the HFD group were higher than those of the ND group (P<0.05), indicating that feeding a high-fat diet accelerated the formation of obesity in mice; compared with the HFD group, when there was no difference in weekly energy intake (P>0.05), the final body weight, weight gain and Lee's index of the Orlistat group, H and M intervention groups were significantly reduced (P<0.05). This indicates that high and medium doses of SSE have a certain weight loss effect on HFD obese mice, just like Orlistat.

[0067] When there was no difference in weekly energy intake, the Lee's index of mice in the Orlistat, H, and M groups was significantly reduced, indicating that medium and high doses of SSE, like Orlistat, had a certain weight loss effect on high-fat diet-induced obese mice.

[0068] Example 8

[0069] Blood was collected from mice by enucleation and centrifuged at 12000r / min for 10min at 4℃. The supernatant was obtained to obtain serum. The corresponding Elisa kits were used to measure the levels of serum metabolism-related hormones leptin (LEP), insulin (INS) and neuropeptide Y (NPY).

[0070] Experimental results:

[0071] like Figure 2 As shown in the figure, the level of INS in the serum of the HFD group was significantly higher than that of the ND group, while the level of INS in the Orlistat group and the H group was lower than that of the HFD group. This indicates that SSE can reduce the level of INS and improve obesity to a certain extent.

[0072] like Figure 3 As shown in the figure, the level of serum LEP in the HFD group was significantly higher than that in the ND group, while the level of LEP in the Orlistat group, H group, and M group was lower than that in the HFD group in a dose-dependent manner. This indicates that SSE can reduce the level of LEP and improve obesity to a certain extent.

[0073] like Figure 4 As shown in the figure, the level of NPY in the serum of the HFD group was significantly higher than that of the ND group, while the level of NPY in the H group was lower than that of the HFD group. This indicates that SSE can reduce the level of NPY and improve obesity to a certain extent.

[0074] Example 9

[0075] The mice in Example 8 were dissected, and samples of the heart, liver, kidney, testis, epididymal white adipose tissue (Epi-WAT), perirenal white adipose tissue (Per-WAT), and brown adipose tissue (BAT) were weighed and collected. All collected samples were stored in a refrigerator at -80°C until analysis.

[0076] Experimental results:

[0077] like Figure 5As shown in the figure, the liver weight of mice in the HFD group increased significantly compared with that in the ND group, while the liver weight of mice in the Orlistat group, H group, and M group decreased significantly compared with that in the HFD group. This indicates that medium and high doses of SSE, like Orlistat, can improve liver damage induced by a high-fat diet to a certain extent and help reduce the obesity of mice.

[0078] like Figure 6 As shown in the figure, the Per-WAT index of the HFD group mice was significantly higher than that of the ND group. The Per-WAT index of the Orlistat group, H group, and M group mice was significantly lower than that of the HFD group mice. This indicates that medium and high doses of SSE, like Orlistat, can improve the liver damage induced by a high-fat diet to a certain extent, which is beneficial to reduce the obesity of mice.

[0079] like Figure 7 As shown in the figure, the Epi-WAT index of mice in the HFD group was significantly higher than that in the ND group. However, the Epi-WAT index of mice in the Orlistat group, H group, and M group was significantly lower than that in the HFD group. This indicates that medium and high doses of SSE, like Orlistat, can improve liver damage induced by a high-fat diet to a certain extent, and are beneficial for reducing the obesity of mice.

[0080] like Figure 8 As shown in the figure, the BAT index of mice in the HFD group was significantly lower than that in the ND group. However, the BAT index of mice in the Orlistat group, H group, and M group was significantly higher than that in the HFD group. This indicates that medium and high doses of SSE, like Orlistat, can improve liver damage induced by a high-fat diet to a certain extent and help reduce the obesity of mice.

[0081] Example 10

[0082] For the mice in Example 9, one liver was randomly selected from each group, and part of the tissue was taken. The tissue was immediately immersed in a newly prepared 4% paraformaldehyde fixative and fixed at room temperature. After the tissue was fixed, it was rinsed with running water, dehydrated with gradient alcohol, and the dehydrated tissue was embedded in paraffin. When the tissue wax block was completely hardened, it was sliced ​​with a thickness of 5 μm. The paraffin sections were stained with hematoxylin and eosin (H&E) and oil red O (ORO) solutions, and the cell morphology was observed under an optical microscope after rinsing and sealing.

[0083] Experimental results:

[0084] like Fig. 9As shown, no pathological signs were observed in the ND group cells, the hepatocyte nuclei were large and round, the cytoplasm was rich, the lipid droplet content was very small, and the liver plates were arranged in an orderly manner. In the HFD group, typical cell pathological phenomena were clearly observed. Compared with the HFD group, the pathological changes of the cells in the Orlistat group were basically restored to the same level as the ND group; the pathological changes of the cells in the H and M groups were significantly improved, reducing the inflammatory infiltration of stem cells, the enlargement of lipid droplets, and the number of cell necrosis; the symptoms of the cells in the L group were slightly alleviated, but the pathological phenomena were similar to those in the HFD group. This shows that medium and high doses of SSE, like Orlistat, can improve and reduce liver inflammation and reduce fat accumulation to a certain extent, which is beneficial to reducing the obesity of mice.

[0085] like Fig.10 As shown in the figure, no pathological state was observed in the ND group, with very few adipocytes and very little lipid droplet accumulation, while the HFD group had obvious adipocyte distribution and accumulation, and a large amount of abnormal lipid deposition. Compared with the HFD group, the lipid content in the Orlistat group and the H group was reduced to basically the same as the ND group. The lipid content in the H, M, and L groups decreased and decreased with the decrease in SSE dose. This shows that medium and high doses of SSE, like Orlistat, can reduce liver inflammation and fat accumulation to a certain extent, which is beneficial to reduce the obesity of mice.

[0086] According to the above content, it can be seen that the willow flower extract involved in the present invention can effectively reduce the body weight and Lee's index of mice, inhibit the accumulation of fat and prevent the formation of fatty liver, and significantly reduce the INS level, LEP level and NPY level in serum. In general, the willow flower extract can reduce liver inflammation and fat accumulation to a certain extent, which is beneficial to reduce the obesity of mice.

[0087] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A willow flower extract, characterized in that The extract is a willow flower extract obtained by extracting willow flower powder with an organic solvent.

2. The extract according to claim 1, characterized in that The organic solvent is ethanol.

3. The use according to claim 2, characterized in that The ratio of the willow flower powder to ethanol is 1g:15-25mL.

4. The use according to claim 1, characterized in that The extraction temperature is 85-95°C.

5. The use according to claim 1, characterized in that After the extraction is completed, the mixture is concentrated by rotary evaporation.

6. Use of the extracts according to claims 1 to 5 in the preparation of fat-reducing products.

7. Use of the extracts according to claims 1 to 5 in the preparation of a drug for preventing obesity caused by a high-fat diet.

8. Use of the extracts as claimed in claims 1 to 5 in the preparation of products for regulating serum hormones.

9. The use according to claim 8, characterized in that The serum hormones include leptin, insulin and neuropeptide Y.

10. Use of the extract according to claims 1 to 5 in the preparation of a liver protection product.

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