CN111227233B

CN111227233B - Application of luffa powder in improving hyperchain-branched chain amino acid blood disease and inhibiting obesity

Info

Publication number: CN111227233B
Application number: CN202010135111.9A
Authority: CN
Inventors: 陈芳; 张璐; 胡小松; 廖小军
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2020-03-02
Filing date: 2020-03-02
Publication date: 2022-07-08
Anticipated expiration: 2040-03-02
Also published as: CN111227233A

Abstract

The invention relates to the technical field of food nutrition, in particular to application of towel gourd powder in improving hyper-branched chain amino acidemia and inhibiting obesity. In order to achieve the purpose, the invention provides the following technical scheme: application of Luffa cylindrica powder for improving hyperchain-branched-chain-amino-acid blood disease and inhibiting obesity is provided. In the invention, the towel gourd powder is proved to inhibit the occurrence of obesity by improving the hyper-branched chain amino acidemia. In the invention, the preparation method of the towel gourd powder comprises the following steps: cleaning and slicing fructus Luffae, freeze drying at low temperature, and micronizing. The method can maximally retain nutritional components of fructus Luffae. The results show that the luffa powder can up-regulate the expression of branched chain amino acid catabolism enzyme gene transcription level in fat tissues and livers of fat mice, promote in vivo branched chain amino acid catabolism, improve hyper-branched chain amino acidemia, inhibit obesity and insulin resistance, and can be used as a dietary nutrient component for inhibiting obesity generation by improving the hyper-branched chain amino acidemia.

Description

Application of luffa powder in improving hyper-branched chain amino acidemia and inhibiting obesity

Technical Field

The invention relates to the technical field of food nutrition, in particular to application of towel gourd powder in improving hyper-branched chain amino acidemia and inhibiting obesity. Many clinical studies have shown that hyper-branched aminoacidemia may be a diagnostic index for the prevention or treatment of obesity and its metabolic syndrome. Animal experiments prove that the towel gourd powder prepared by low-temperature freeze drying can promote the catabolism of redundant branched-chain amino acids in vivo by up-regulating the expression of the branched-chain amino acid catabolic enzyme, improve the hyper-branched-chain amino acid blood disease, further relieve metabolic diseases such as obesity and the like, and can be used as a dietary additive or a dietary structural component for improving the metabolic disorder of the branched-chain amino acids and inhibiting the occurrence of obesity.

Background

Epidemiology has shown that the incidence of obesity is rapidly rising in developed and developing countries, and the disease is in a state of youthfulness, and complications caused by obesity are diverse. Obesity caused by modern dietary habits (high fat and low dietary fiber) can cause imbalance of human body physiological homeostasis (lipid metabolism, sugar metabolism and amino acid metabolism) and is accompanied with diseases such as hyperlipidemia, hyperglycemia, type II diabetes, cardiovascular and cerebrovascular diseases, certain cancers and the like.

Many studies have demonstrated that high or low levels of branched chain amino acids in the body can be used to diagnose the course of obesity and its metabolic diseases. Branched-chain amino acids (BCAAs) include valine (Val), leucine (Leu), and isoleucine (Ile), which are essential amino acids for human body. Food is the main source of BCAA in human body, and BCAA catabolism in the body keeps dynamic balance under normal physiological conditions. BCAAs catabolism mainly occurs in mitochondria of adipose tissues and liver tissues, the first step is the branched-chain alpha-keto acid catalyzed by branched-chain amino acid transaminase (BCATm), the second step is the branched-chain alpha-keto acid dehydrogenase kinase (BCKDK) complex which is further oxidized and decarboxylated and enters the tricarboxylic acid cycle to generate NADH, CO₂And the like in the final product. A large number of researches indicate that long-term high-fat diet intake causes in-vivo BCAA metabolic disorder, and the specific manifestations are that BCAA catabolic enzyme expression is reduced, branched chain amino acid in blood is excessively accumulated, and then diseases such as obesity and insulin resistance are induced or aggravated. In recent years, people pay attention to diet therapy to prevent and relieve obesity and metabolic complications thereof through various ways, and it is important to find a diet or a diet composition which is effective in improving related diseases.

Luffa cylindrica, also called Mian melon, Bu melon and Man melon, is an annual climbing herb plant of Luffa genus of Cucurbitaceae family, and is widely planted in the world. The towel gourd can be used as both medicine and food, and fresh and tender fleshy fruits of the towel gourd belong to bottle gourd fruits, are rich in dietary fibers, vitamins, polyphenol, flavonoid, saponin, triterpenes, oleanolic acid, carotene, chlorophyll and other nutrient elements, can be used as a Chinese herbal medicine for clearing heat and removing toxicity, relieving cough and fever, and treating dyspnea and subcutaneous swelling, and is a vegetable food with good health efficacy. The loofah powder prepared in the patent contains 51.47% of carbohydrate, 21.75% of protein, 0.7% of fat, 5.34% of ash and 20.74% of dietary fiber according to the measurement of nutrient components, has the energy density of 3.04Kal/g, and has good characteristics of low fat and low calorie. The seeds of the aged and dried luffa fruits are luffa seeds, also called black cowberry seeds, which are flat and oval, and have grey black or black seed coats and slightly bitter taste. The luffa seeds are rich in various natural lipids and other nutrient substances, and are reported to contain about 24-39% of fatty oil, mainly linoleic acid, palmitic acid, stearic acid, oleic acid and the like, and have the effects of clearing damp-heat, tonifying qi, benefiting middle-jiao, relaxing bowels and expelling parasites. Chinese patent 2012106544160.3 discloses the use of luffa seeds for preventing and alleviating diseases related to blood lipid elevation. Chinese patent 201510577827.3 provides the use of luffa extract for the prevention and treatment of obesity. However, at present, no report is found on the application of the towel gourd in inhibiting the occurrence of obesity by improving the hyperchain aminoacemia.

Disclosure of Invention

The invention aims to provide application of towel gourd powder in improving hyper-branched chain amino acidemia and inhibiting obesity.

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

application of Luffa cylindrica powder for improving hyperchain-branched-chain-amino-acid blood disease and inhibiting obesity is provided.

The preparation method of the towel gourd powder comprises the following steps:

1) cleaning and slicing loofah: picking fresh towel gourd, cleaning and cutting into slices;

2) and (3) low-temperature freeze drying: pre-freezing the towel gourd slices in the step 1 at the temperature of between 20 ℃ below zero and 80 ℃ below zero for 24 to 48 hours, and then freezing and drying the towel gourd slices at the temperature of between 60 ℃ below zero and 80 ℃ below zero for 48 to 72 hours;

3) superfine grinding: and 2, carrying out superfine grinding on the freeze-dried towel gourd slices in the step 2, and then sieving with a 60-mesh sieve to obtain towel gourd powder.

Wherein, in the step 1, the picked fresh towel gourd is cleaned and cut into slices with the thickness of 2-3 mm.

Wherein, in the step 2, the towel gourd slices in the step 1 are pre-frozen at-80 ℃ for 48 hours, and then are freeze-dried at-80 ℃ for 72 hours.

Wherein, the preparation method of the towel gourd powder further comprises the following steps:

4) and (4) storing the towel gourd powder prepared in the step (3) at the temperature of minus 20 ℃ in a dark place.

Wherein, the towel gourd powder obtained in the step 3 is mixed with sterile normal saline to be 0.2g/mL before use to prepare a towel gourd suspension.

Compared with the prior art, the invention has the beneficial effects that: in the invention, the towel gourd powder is proved to inhibit the occurrence of obesity by improving the hyper-branched chain amino acidemia.

In the invention, the preparation method of the towel gourd powder comprises the following steps: cleaning and slicing fructus Luffae, freeze drying at low temperature, and micronizing. The method can maximally retain nutritional components of fructus Luffae.

The results show that the luffa powder can up-regulate the expression of branched chain amino acid catabolism enzyme gene transcription level in fat tissues and livers of fat mice, promote in vivo branched chain amino acid catabolism, improve hyper-branched chain amino acidemia, inhibit obesity and insulin resistance, and can be used as a dietary nutrient component for inhibiting obesity generation by improving the hyper-branched chain amino acidemia.

Drawings

FIG. 1 shows the effect of Luffa cylindrica Roem on the expression of branched-chain amino acid catabolic enzyme genes in adipose tissue and liver tissue of mice on high fat diet. Wherein, the lower case letters indicate that the difference comparison is carried out at a p-0.05 level, the letters are different to indicate that the difference between different treatment groups is significant (p <0.05), and the letters are the same to indicate that the difference between different treatment groups is not significant (p > 0.05);

FIG. 2 shows the effect of Luffa cylindrica powder on the level of branched chain amino acids in serum of mice on a high-fat diet. Wherein, the lower case letters indicate that the difference comparison is carried out at the level of p ═ 0.05, the letters are different to indicate that the difference between different treatment groups is significant (p <0.05), and the letters are the same to indicate that the difference between different treatment groups is not significant (p > 0.05);

fig. 3 and 4 show the effect of luffa powder on body weight and fat distribution in mice on high-fat diet. Wherein, the lower case letters indicate that the difference comparison is carried out at a p-0.05 level, the letters are different to indicate that the difference between different treatment groups is significant (p <0.05), and the letters are the same to indicate that the difference between different treatment groups is not significant (p > 0.05);

FIG. 5 shows the effect of Luffa cylindrica powder on lipid accumulation in adipose tissues of mice on high-fat diet;

FIG. 6 shows the effect of Luffa cylindrica powder on liver tissue lipid accumulation in mice with high-fat diet.

Detailed Description

The following further describes embodiments of the present invention with reference to the drawings.

The preparation method of the towel gourd powder comprises the following steps:

1. cleaning and slicing loofah: picking fresh towel gourd, cleaning and slicing;

preferably, in the step 1, the picked fresh towel gourd is cleaned and cut into slices with the thickness of 2-3 mm.

2. And (3) low-temperature freeze drying: pre-freezing the towel gourd slices in the step 1 at the temperature of between 20 ℃ below zero and 80 ℃ below zero for 24 to 48 hours, and then freezing and drying the towel gourd slices at the temperature of between 60 ℃ below zero and 80 ℃ below zero for 48 to 72 hours.

Preferably, in step 2, the luffa slices in step 1 are pre-frozen at-80 ℃ for 48h, and then freeze-dried at-80 ℃ for 72 h.

3. Ultra-fine crushing: and 2, carrying out superfine grinding on the freeze-dried towel gourd slices in the step 2, and then sieving the slices through a 60-mesh sieve to obtain the towel gourd powder.

Preferably, the preparation method of the luffa powder further comprises the following steps:

4. and (4) storing the towel gourd powder prepared in the step (3) at the temperature of minus 20 ℃ in a dark place.

Preferably, the loofah powder obtained in the step 3 is mixed with sterile normal saline to be 0.2g/mL to prepare a loofah suspension before use.

The loofah powder prepared by the method can retain the nutritional ingredients of the loofah to the maximum extent.

The influence of the luffa powder on the expression of branched-chain amino acid catabolic enzyme genes in adipose tissues and liver tissues of high-fat diet mice, the influence of the luffa powder on the level of branched-chain amino acids in serum of the high-fat diet mice, the influence of the luffa powder on body weight and fat distribution of the high-fat diet mice, the influence of the luffa powder on fat tissue lipid accumulation of the high-fat diet mice, and the influence of the luffa powder on liver tissue lipid accumulation of the high-fat diet mice:

1. preparation of towel gourd intervention substance

Picking fresh fructus Luffae, cleaning, cutting into slices with thickness of 2-3mm, pre-freezing at-80 deg.C for 48 hr, freeze-drying at-80 deg.C for 72 hr, micronizing, sieving with 60 mesh sieve to obtain fructus Luffae powder, and storing at-20 deg.C in dark place. Before use, sterile normal saline is mixed into 0.2g/mL luffa powder suspension, and the stomach filling dose is 10 mu L/g mouse weight.

2. Animal experiments

1)30 healthy C57BL/6J male mice (available from Beijing Wintolite, Inc.) aged 5 weeks were randomized into 3 groups of 10 mice each, one cage of 5, one week after acclimation. The first group was fed control diet (D12450B, 3.85Kcal/g, available from Research Diets, USA) and gavage of equal doses of saline, i.e., control group; the second group was fed with high fat diet (D12492, 5.24Kcal/g, available from Research Diets, USA) and gavage of equal doses of physiological saline, i.e., high fat group; and feeding high-fat feed and intragastrically feeding 0.2g/mL loofah powder suspension, namely the dried silk melon pre-group. Experimental time 12 weeks, 8 am each day: 30-9: 30 gavage, during the experiment the mice had free access to feed and water.

2) And (3) weighing the weight of each group of mice after the experiment is finished, collecting the serum, the liver tissue and the epididymis adipose tissue of each group of mice, detecting the serum sample by total triglyceride TG, total cholesterol TC, high-density lipoprotein HDL, low-density lipoprotein LDL, fasting blood glucose Glu and fasting insulin INS as blood biochemical indexes, and detecting the content of free fatty acid FFA in the serum by using an ELISA kit. The insulin resistance index HOMA-IR (HOMA-IR ═ (Glu (mmol/L) × INS (mIU/L))/22.5) was calculated for each group of mice. Part of liver and adipose tissue are used for pathological examination and histological analysis, and part of liver and adipose tissue are used for transcription level expression analysis of branched chain amino acid catabolic enzyme genes.

The results obtained were as follows:

FIG. 1 shows the effect of Luffa cylindrica Roem on the expression of branched-chain amino acid catabolic enzyme genes in adipose tissue and liver tissue of mice on high fat diet. Wherein, the lower case letters indicate that the difference comparison is carried out at the level of p ═ 0.05, the letters are different to indicate that the difference between different treatment groups is significant (p <0.05), and the letters are the same to indicate that the difference between different treatment groups is not significant (p > 0.05).

Fig. 1 shows that high fat diet leads to significant reduction of more than 50% of mRNA level expression of BCAA catabolism key enzymes BCATm and BCKDK in mouse adipose tissues compared with control group, and mRNA level expression of BCATm and BCKDK in mice of towel gourd intervention group can be restored to control group level; the mRNA level expression of BCATm in the liver tissue of the high-fat diet mouse is also obviously reduced by about 50 percent, and the mRNA level expression of the BCATm in the towel gourd powder dry-treatment group is obviously recovered and is basically the same as that in the control group; although the expression level of the mRNA of the BCKDK in the liver of the mouse with high fat diet is approximately two times higher than that of the control group, the expression level of the BCKDK in the liver of the mouse with the towel gourd intervention group has no significant difference compared with the control group and the high fat group; therefore, the intervention of the luffa powder can obviously improve the gene expression of branched chain amino acid catabolic enzymes in adipose tissues and liver tissues of high-fat diet mice.

FIG. 2 shows the effect of Luffa cylindrica powder on the level of branched chain amino acids in serum of mice on a high-fat diet. Wherein, the lower case letters indicate that the difference comparison is carried out at the level of p ═ 0.05, the letters are different to indicate that the difference between different treatment groups is significant (p <0.05), and the letters are the same to indicate that the difference between different treatment groups is not significant (p > 0.05).

Fig. 2 shows that the contents of valine, leucine and isoleucine in the serum of the high-fat diet mouse are respectively and remarkably increased by about 41%, 38% and 65% compared with the control group, and the indexes of the towel gourd intervention group mouse are only respectively increased by about 22%, 9% and 20%; in general, the serum total branched chain amino acid level of the control group mouse is 46.32 +/-3.38 mu g/mL, the index in the serum of the high-fat group mouse is obviously increased by about 44-66.61 +/-3.93 mu g/mL, and the index in the serum of the towel gourd intervention group mouse is only increased by about 16-53.83 +/-1.98 mu g/mL; therefore, the mouse with high fat diet has hyper-branched chain amino acidemia, the BCAA level in vivo is obviously increased, and the intervention of the towel gourd powder can obviously relieve the hyper-branched chain amino acidemia.

Fig. 3 and 4 show the effect of luffa powder on body weight and fat distribution in mice on high-fat diet. Wherein, the lower case letters indicate that the difference comparison is carried out at the level of p ═ 0.05, the letters are different to indicate that the difference between different treatment groups is significant (p <0.05), and the letters are the same to indicate that the difference between different treatment groups is not significant (p > 0.05).

Fig. 3 and 4 show that compared with the control group, the weight of the high-fat diet mice is obviously increased, the whole body has obvious fat distribution, and the intervention of the towel gourd powder can obviously reduce the weight increase of the high-fat diet mice and relieve the fat distribution of the whole body.

FIG. 5 shows the effect of Luffa cylindrica powder on lipid accumulation in adipose tissue of mice on high-fat diet.

Fig. 5 shows that compared with the control group, the cell volume in the adipose tissue of the high-fat diet mouse is obviously increased, the cell size is unequal, the lipid accumulation is serious, and the lipid accumulation in the adipose cells of the high-fat diet mouse can be obviously reduced by the intervention of the luffa powder.

FIG. 6 shows the effect of Luffa cylindrica Roem on lipid accumulation in liver tissue of mice on high-fat diet.

Fig. 6 shows that compared with the control group, the liver tissues of the mice with high-fat diet have different lipid droplet sizes and serious lipid accumulation, and the intervention of the towel gourd powder can obviously reduce the lipid accumulation in the liver cells of the mice with high-fat diet.

Table 1 shows the effect of luffa on biochemical indicators of blood in mice fed with high fat:

	control group	High fat group	Dried cucumber pre-treating group
				FFA(mmol/L)	0.35±0.02^b	0.46±0.01^a	0.37±0.01^b
TG(mmol/L)	0.39±0.01^b	0.45±0.24^a	0.39±0.05^b
				TC(mmol/L)	3.19±0.19^b	4.63±0.24^a	3.65±0.06^b
HDL(mmol/L)	2.54±0.17^b	3.44±0.12^a	2.88±0.09^b
				LDL(mmol/L)	0.56±0.001^b	0.92±0.13^a	0.62±0.05^b
Glu(mmol/L)	5.20±0.08^c	8.40±0.40^a	6.06±0.36^b
				INS(mIU/L)	11.45±1.31^b	18.38±1.66^a	13.02±0.67^b
HOMA-IR	2.65±0.34^b	6.92±0.91^a	3.54±0.38^b

Table 1 shows that the towel gourd powder can obviously reduce the lipid level (FFA, TG and TC) in the serum of the high-fat diet mouse, reduce the LDL level in the serum, reduce the blood sugar content and the insulin content and improve the insulin resistance of the high-fat diet mouse.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and their concepts should be equivalent or changed within the technical scope of the present invention.

Claims

1. Application of fructus Luffae powder in preparing non-therapeutic nutritional product for improving hyperchain-branched chain amino acid blood disease is provided.

2. The use of claim 1, wherein: the preparation method of the towel gourd powder comprises the following steps:

3) superfine grinding: and 2, carrying out superfine grinding on the freeze-dried towel gourd slices in the step 2, and then sieving the slices through a 60-mesh sieve to obtain the towel gourd powder.

3. Use according to claim 2, characterized in that:

in the step 1, picked fresh towel gourds are cleaned and cut into slices with the thickness of 2-3 mm.

4. Use according to claim 2, characterized in that:

in the step 2, the towel gourd slices in the step 1 are pre-frozen at-80 ℃ for 48 hours, and then are frozen and dried at-80 ℃ for 72 hours.

5. The use of claim 2, wherein:

the preparation method of the towel gourd powder further comprises the following steps:

6. The use of claim 2, wherein:

and (4) mixing the towel gourd powder obtained in the step (3) with sterile normal saline to prepare a towel gourd suspension at a concentration of 0.2g/mL before use.