CN115463188A

CN115463188A - Application of asparagus cochinchinensis water extract in preparation of preparation for preventing or treating obesity

Info

Publication number: CN115463188A
Application number: CN202211136406.3A
Authority: CN
Inventors: 邱景富; 陈承志; 邹镇; 罗诗月; 唐世鑫
Original assignee: Chongqing Medical University
Current assignee: Chongqing Medical University
Priority date: 2022-09-19
Filing date: 2022-09-19
Publication date: 2022-12-13

Abstract

The invention belongs to the technical field of medicine preparation, and particularly relates to application of an asparagus cochinchinensis water extract in serving as or preparing a preparation for preventing or treating obesity. The liquid extracted from the radix asparagi water body can remarkably reduce weight gain and fat tissue accumulation, improve blood fat and liver dysfunction, and inhibit inflammation and lipogenesis of liver and fat tissue.

Description

Application of asparagus cochinchinensis water extract in preparation of preparation for preventing or treating obesity

Technical Field

The invention belongs to the technical field of medicine preparation, and particularly relates to application of an asparagus cochinchinensis water extract in serving as or preparing a preparation for preventing or treating obesity.

Background

Obesity is a chronic metabolic disease caused by the interaction of multiple factors such as heredity and environment, such as excessive fat accumulation in body, abnormal distribution and increased body constitution, and is diagnosed with Body Mass Index (BMI) of not less than 30kg/m ² . Obesity can promote muchThe occurrence or development of chronic diseases, such as cardiovascular diseases, diabetes, partial cancers, osteoarthritis, and the like. As the prevalence of overweight and obesity continues to increase worldwide, concerns over health risks associated with this increasingly worsening problem are exacerbated. The medicine which can be clinically used for treating the obesity has flexible fingers at present, and the safety and the curative effect are not yet determined.

Efforts are constantly being made to explore strategies for preventing and managing obesity, and improving dietary habits, increasing physical activity, participating in weight loss surgery or taking weight loss drugs are the main options. However, a good drug target does not exist at present, and in the development process of weight-reducing drugs, many drugs act on nerve feeding centers, such as phentermine, orlistat and liraglutide 3.0, which have many side effects, such as nausea, dry mouth, difficulty in sleeping and the like.

Therefore, there is a need for a more effective, safer and less side-effect weight-loss drug.

Disclosure of Invention

In view of the above problems, an object of the present invention is to provide an extract of Asparagus cochinchinensis in the form of an aqueous solution, or in the form of a preparation for preventing or treating obesity. The radix asparagi water extract can effectively reduce the body index of obesity and can effectively prevent or treat obesity.

In order to achieve the purpose, the following technical scheme can be adopted:

the invention provides an application of an asparagus cochinchinensis water extract in preparing or preparing a preparation for preventing or treating obesity.

In still another aspect, the present invention provides a preparation for preventing or treating obesity, which comprises at least an aqueous extract of Asparagus cochinchinensis.

The beneficial effects of the invention at least comprise: according to the embodiment of the invention, the liquid extracted from the radix asparagi water body can obviously reduce weight gain and fat tissue accumulation, improve blood fat and liver dysfunction, and inhibit inflammation and lipogenesis of the liver and fat tissue.

Drawings

FIG. 1 is a weight-growth curve obtained for the body weight of mice;

FIG. 2 is a graph of the total body weight gain of mice at the end of the treatment after 10 weeks;

FIG. 3 shows the total weight of white adipose tissues in mice;

FIG. 4 shows the fat index profile of mice;

FIG. 5 is the Triglyceride (TG) content in the plasma of mice;

FIG. 6 shows Total Cholesterol (TC) levels in mouse plasma;

FIG. 7 is a graph showing the low density lipoprotein (LDL-C) content in mouse plasma;

FIG. 8 shows the high density lipoprotein (HDL-C) content in mouse plasma;

FIG. 9 shows the alanine Aminotransferase (ALT) content in mouse plasma;

FIG. 10 shows the levels of aspartate Aminotransferase (AST) in the plasma of mice;

FIG. 11 is a representative picture of dissected mouse abdomen;

FIG. 12 is a representative picture of Epi-WAT tissue hematoxylin and eosin (H & E) staining;

FIG. 13 is a representative picture of hematoxylin and eosin (H & E) staining of liver tissue;

FIG. 14 is a representative picture of liver tissue oil red O staining;

FIG. 15 is the average adipocyte area after H & E staining of adipose tissue;

FIG. 16 is a graph of the effect of Asparagus cochinchinensis on liver tissue inflammation genes in high fat diet obese mice;

FIG. 17 is a graph of the effect of Asparagus cochinchinensis on the lipid metabolism gene of adipose tissue in high-fat diet obese mice;

FIG. 18 is a graph of the effect of Asparagus cochinchinensis on hepatic lipid metabolism genes in high fat diet obese mice.

Detailed Description

The examples are given for the purpose of better illustration of the invention, but the invention is not limited to the examples. Therefore, those skilled in the art should make insubstantial modifications and adaptations to the embodiments of the present invention in light of the above teachings and remain within the scope of the invention.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the disclosure. Unless the context has a significantly different meaning, the singular forms of expressions include the plural forms of expressions. As used herein, it is understood that terms such as "comprising," "having," "including," and the like are intended to indicate the presence of features, numbers, operations, components, parts, elements, materials, or combinations thereof. The terms of the present invention are disclosed in the specification and are not intended to exclude the possibility that one or more other features, numbers, operations, components, parts, elements, materials or combinations thereof may be present or may be added. As used herein, "/" can be interpreted as "and" or "depending on the situation.

In some embodiments, in the above application, the obesity is prevented or treated by reducing the accumulation of adipose tissues, i.e., the radix asparagi water extract can reduce the accumulation of adipose tissues in the body, thereby improving or preventing obesity.

In some embodiments, the above application can prevent or treat obesity by improving blood lipid and liver dysfunction, i.e., radix asparagi water extract can improve blood lipid and liver dysfunction of body.

In some embodiments, the obesity is prevented or treated by suppressing inflammation of the liver, i.e., the extract solution of Asparagus cochinchinensis can suppress inflammation of the liver.

In some embodiments, the obesity is prevented or treated by suppressing liver lipogenesis in the above application, i.e., the extract of radix asparagi water can suppress liver lipogenesis.

In some embodiments, in the above formulation, the formulation may be a medicament or a food. Specifically, the asparagus aqueous extract can be prepared into a medicament with larger medicament ratio, a health-care food with health-care function and the like.

In some embodiments, the preparation may further include some pharmaceutical excipients or diluents in addition to the radix asparagi water extract, the pharmaceutical excipients or diluents may be used to prepare the drug into various dosage forms, such as tablets, emulsions, aqueous solutions, etc., and the excipients or diluents may be added according to specific needs.

It should be noted that the liver is a major metabolic organ of the human body and plays an important role in lipid metabolism, such as endogenous fat production, in vivo fat transport and storage. There is increasing evidence that high fat or high sugar diets alter genes involved in fatty acid synthesis and metabolism in the liver, and that anti-obesity candidate genes may exert anti-obesity effects by modulating the expression of these genes. In addition, adipose tissue is an extremely important secretory organ as well as an energy storage organ, and plays an important role in the regulation of energy and metabolism in the body, like the liver. The disorder of adipocyte function is closely related to the occurrence of obesity and related diseases. An increase in the number or volume of adipocytes is the pathological basis of obesity, and thus inhibition of adipocyte differentiation is an important direction in the development of current anti-obesity drugs.

It should be noted that high fat diet changes the genes related to fatty acid synthesis and metabolism in the liver, and in the present example, after the radix asparagi decoction is taken, the up-regulation of the genes SREBP-1c, FAS, ACC and SCD-1 which regulate fatty acid synthesis in the liver caused by high fat diet is reduced. The reduction in expression of these genes may at least partially account for the improvement in steatosis observed following cochinchnese asparagus administration, such as H & E and oil red O staining results. In addition to liver tissue, adipose tissue is also essential for controlling system homeostasis. During excessive nutrient intake, adipocytes will grow and proliferate; in some further embodiments of the invention, the H & E results show a significant improvement in adipocyte proliferation and increase of high fat diet after administration of the asparagus cochinchinensis aqueous extract, significantly up-regulating the expression of genes involved in induction of lipid oxidation in adipose tissue; from these results, it can be seen that Asparagus cochinchinensis can reduce fat deposition in mice fed a high-fat diet.

It should be noted that the aqueous extract of Asparagus Cochinchinensis in the above applications and drugs is obtained by extracting Asparagus Cochinchinensis by water extraction method known in the art. Specifically, asparagus cochinchinensis (Lour.) Merr, which is a perennial climbing herb of Asparagus of Liliaceae, is sweet and bitter in taste and cold in nature as dry root tuber. In China, 24 asparagus plants are widely distributed in various provinces of south and north, have more than two thousand years of use history and are one of the bulk medicinal materials exported traditionally. Asparagus cochinchinensis is recorded in the book Shen nong Ben Cao Jing, the book Bie Lu, the book Ben Cao Jing Ji Yi, the book Ben Cao Zheng Yi and the book Ben Cao gang mu, etc. Is widely applied to the treatment of common diseases such as lung dryness, dry cough, cough with sticky phlegm, cough due to asthenia, vexation, insomnia, internal heat, diabetes, constipation due to intestinal dryness, cancer prevention and cancer treatment, and the like by the medical field. Modern pharmacological experiment research shows that the plant extract has the functions of resisting tumor, resisting inflammation, improving immunity, resisting oxidation, resisting senility, resisting ulcer, resisting diarrhea, resisting thrombosis, etc. However, the pharmacological activity of asparagus cochinchinensis is not deeply studied, and reports of asparagus cochinchinensis on obesity have been searched so far.

It is noted that the asparagus cochinchinensis aqueous extract belongs to natural medicines, and is used in traditional medicine, so that the toxicity caused by the developed medicines does not need to be paid deep attention; in some embodiments, the asparagus cochinchinensis aqueous extract can significantly improve the weight gain of obese mice induced by feeding high fat feed containing 60% fat, reduce the accumulation of triglyceride in plasma, and improve the lipid metabolism disorder of liver and adipose tissue.

For a better understanding of the present invention, the following further illustrates the contents of the present invention with reference to specific examples, but the contents of the present invention are not limited to the following examples.

In the following examples, the preparation of the asparagus extract was carried out according to the following steps: selecting high-quality radix asparagi from Guizhou producing area of China, weighing about 100g of radix asparagi root, and soaking in cold water for 20-30min; then, preparing the water extract of the radix asparagi according to the following steps: adding water for 2 times, wherein the water is 600mL for the first time and 400mL for the second time; each time, heating slowly for 30min after boiling; then, the decoction obtained in the two steps is cooled and filtered through four layers of gauze to remove residues. The filtered decoction was then concentrated to 100mL using a rotary evaporator. Then, obtaining an asparagus root extracting solution with the final concentration of 1 g/mL; sterilizing with high pressure steam.

In the following examples, mouse model establishment was performed as follows: taking 36C 57BL/6J male mice with age of 4 weeks and weight of 12-16g, keeping the room temperature at 23 + -1 deg.C and humidity at 50 + -60%; the light and dark cycle is 12 hours, and clean food and water can be obtained at will for training; the experimental method comprises the following steps: one week after training, mice were randomly divided into four groups of 9 mice, a normal diet group (ND), a normal diet plus asparagus group (ND + Ac), a high fat diet group (HFD) and a high fat diet plus asparagus group (HFD + Ac); 60% fat-energized high fat feed is used; the oral asparagus cochinchinensis water extract of the treatment group is 1g/kg/d, the physiological saline with the same volume is orally taken by a blank control group, and the weight of each mouse is weighed once every three days; after 10 consecutive weeks of treatment, all mice were fasted for 6h and sacrificed after anesthesia with ketamine (80 mg/kg) and thiazine (6 mg/kg); liver tissue, perirenal white adipose tissue (Per-WAT), epididymal white adipose tissue (Epi-WAT) and blood were collected from each mouse for subsequent analysis.

In the following examples, data are presented as mean ± standard deviation (SEM), and independent sample T-tests were used to assess differences between the two groups, all graphs being created by graphpadprism 8.0.

EXAMPLE 1 Biochemical parameter evaluation

The weight-growth curve obtained by weighing the mouse once every three days is shown in fig. 1; the total body weight increased at the end of the treatment after 10 weeks, as shown in figure 2.

Blood from the eyeballs of the mice after the sacrifice under anesthesia was collected, centrifuged at 3000 Xg for 15min at 4 ℃ to separate plasma, triglyceride (TG), total Cholesterol (TC), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), alanine Aminotransferase (ALT) and aspartate Aminotransferase (AST) were measured, and the samples were stored at-80 ℃ for further analysis, and the results were as follows:

the total weight of white adipose tissues of the mice is shown in figure 3; the fat index of the mice is shown in figure 4; the body weight, adipose tissue weight and fat index of HFD mice were significantly increased compared to ND group. In contrast, asparagus treatment significantly reduced weight gain caused by HFD;

the plasma Triglyceride (TG) content, total Cholesterol (TC) content, low-density lipoprotein (LDL-C) content, high-density lipoprotein (HDL-C) content, alanine Aminotransferase (ALT) content, and aspartate Aminotransferase (AST) content of the mice are shown in FIGS. 5, 7, 8, 9, and 10, respectively; wherein data are expressed as mean ± standard deviation, N =5, <0.05; * P <0.01; * P <0.001; HFD-induced plasma TC, HDL-C, LDL-C and AST levels were significantly elevated (P < 0.05) compared to the ND group. However, the difference between TG and ALT between the two groups was not statistically significant. The ND + Ac group mice had decreased TG levels compared to the ND group, with no other difference between the two groups. For HFD-fed mice, treatment significantly down-regulated TG and AST levels (P < 0.05). In conclusion, the asparagus may partially improve the dyslipidemia and abnormal liver function caused by HFD.

Example 2 histological analysis

To fix the collected liver samples with freshly prepared 4% paraformaldehyde, epididymal white adipose tissue (Epi-WAT) was fixed with poly (methyl calcium), as shown in FIG. 11, epididymal white adipose tissue (Epi-WAT) was highlighted in yellow;

hematoxylin and eosin staining (HE): baking the liver and the white adipose tissue paraffin section in an oven at 65 ℃ for at least 1.5 hours to ensure that the paraffin on the tissue section is fully melted; the sections were then placed in xylene twice to remove the melted paraffin and dehydrated in gradient ethanol. The sections were stained sequentially with hematoxylin and eosin, and then dehydrated with ethanol; after staining, sections were clarified twice in xylene and fixed with neutral resin. Observing the stained section under an optical microscope, wherein the liver section is amplified by 100 times, and the adipose tissue is amplified by 200 times; epi-WAT tissue hematoxylin and eosin (H & E) staining is shown in fig. 12, where the pictures are magnified 200 x, scale bar: 100um; liver tissue hematoxylin and eosin (H & E) staining is shown in fig. 13, with 200 x magnification, scale bar: 100um; black arrows indicate hepatocyte encystment, red arrows indicate hepatic steatosis;

dyeing with oil red O: freezing liver tissue, fixing in 10% formalin for 10min, and washing with distilled water for 2min; washing with 60% isopropyl alcohol solution for 2s; dyeing with oil red O for 8-10min, washing with 60% isopropanol solution for 5-10s; washing with tap water for 2min; and (3) performing counter staining on hematoxylin for 20s, soaking the section in distilled water for 2min after staining, sealing the section with glycerol, and observing the section under an optical microscope. The slice is magnified by 400 times; the liver tissue oil red O staining is shown in fig. 14, the picture is magnified 400 times, and the scale bar: 100um;

after H & E staining of adipose tissue, mean adipocyte area is shown in fig. 15, data are expressed as mean ± standard deviation, N =3,. P <0.05; * P <0.01; * P <0.001;

from the above, it can be seen from fig. 11 that the epididymal fat of the mice fed with the normal diet after the treatment with cochinchnese asparagus root was not significantly changed, but the epididymal fat deposition of the mice fed with the high-fat diet was reduced. Furthermore, it is also seen from fig. 12 and 15 that asparagus treatment significantly reduced adipocyte enlargement and proliferation (P < 0.05) in adipose tissue of high-fat diet mice; h & E staining and oil-red O staining of liver tissue revealed that the hepatocellular microvesicle steatosis was more severe in the HFD group mice than in the ND group (fig. 13); meanwhile, there was severe lipid droplet accumulation in liver tissue of HFD group (fig. 14). Therefore, the asparagus can obviously relieve the hepatic steatosis and the lipid droplet accumulation caused by high-fat diet.

Example 3 analysis of genes for inflammation and lipid metabolism in liver tissue

Determination of inflammatory gene expression in tissues using real-time fluorescent quantitative PCR (RT-PCR): expression of interleukin-1 β (IL-1 β), interleukin-6 (IL-6), tumor necrosis factor (TNF- α) and monocyte marker CD68, results are shown in figure 16, data are expressed as mean ± standard deviation, N =3,. P <0.05; * P <0.01; * P <0.001. From the figure, it can be seen that the expression level of Il-1 beta and CD68 mRNA in the liver of HFD-fed mice is significantly higher than that of ND-fed mice, while the expression level of mouse-related inflammatory genes is significantly reduced after the treatment of Asparagus tuber (P < 0.05);

determination of lipid metabolism gene expression in tissues using real-time fluorescent quantitative PCR (RT-PCR): sterol regulatory element binding protein-1 c (SREBP-1 c), cluster of differentiation 36 (CD 36), fatty Acid Synthase (FAS), carnitine palmitoyl transferase 1 (CPT-1 α), microsomal triglyceride transfer protein (MTP), peroxisome proliferator-activated receptor α (PPAR- α), peroxisome proliferator-activated receptor γ (PPAR- γ), acetyl-CoA carboxylase (ACC), stearoyl-CoA desaturase-1 (SCD-1); as shown in fig. 17, data are expressed as mean ± standard deviation, N =3, { dot over } <0.05; * P <0.01; * P <0.001. As is clear from FIG. 17, HFD significantly reduced the mRNA levels of the genes ATGL, HSL, PGC 1. Alpha., PPAR-gamma, etc. After treatment with Asparagus, the mRNA level of PGC 1. Alpha. In fat was significantly higher than that in the HFD group. These results indicate that asparagus treatment improves the fat metabolism disturbance in HFD fed mice.

Determination of lipid metabolism gene expression in tissues using real-time fluorescent quantitative PCR (RT-PCR): fatty triglyceride lipase (ATGL), hormone Sensitive Lipase (HSL), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1 alpha), carnitine palmitoyl transferase 1 (CPT-1 alpha), microsomal triglyceride transfer protein (MTP), peroxisome proliferator-activated receptor alpha (PPAR-alpha), peroxisome proliferator-activated receptor gamma (PPAR-gamma); results are shown in fig. 18, data are expressed as mean ± standard deviation, N =3, × P <0.05; * P <0.01; * P <0.001. Compared with the ND group, HFD significantly up-regulated the mRNA levels of the adipogenic genes SREBP-1c, MTP, ACC and SCD-1, while Asparagus down-regulated the mRNA levels of SREBP-1c, FAS, ACC and SCD-1. These results indicate that asparagus improves HFD-induced liver lipodystrophy.

In conclusion, the invention shows that the radix asparagi extracting solution has the anti-obesity effect on mice fed with high-fat diet. We have found that the extract of Asparagus cochinchinensis (lour.) Merr) can reduce weight gain, visceral adipose tissue weight and blood lipid level caused by high fat diet. This beneficial effect may be achieved by improving lipid metabolism in the tissue, inhibiting the inflammatory response. In general, our invention suggests the potential of the asparagus extract in preventing and treating obesity, and provides a new possibility for developing more effective and safer weight-reducing drugs.

Finally, the above embodiments are only intended to illustrate the technical solution of the present invention and not to limit the same, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention, which shall be covered by the claims of the present invention.

Claims

1. Application of radix asparagi water extractive solution in preparing preparation for preventing or treating obesity.

2. Use according to claim 1, wherein the prevention or treatment of obesity is by reducing adipose tissue accumulation.

3. The use according to claim 1, wherein the prevention or treatment of obesity is by improvement of blood lipids and liver dysfunction.

4. The use according to claim 1, wherein the prevention or treatment of obesity is by inhibiting inflammation of the liver.

5. Use according to claim 1, wherein the prevention or treatment of obesity is by inhibition of liver lipogenesis.

6. A preparation for preventing or treating obesity is characterized by comprising an asparagus cochinchinensis water extract.

7. The formulation of claim 6, wherein the formulation comprises a medicament or a food.

8. The formulation of claim 6 or 7, wherein the formulation is in a dosage form comprising a tablet, an emulsion, or an aqueous formulation.