Composition for improving fatty liver and application thereof

Abstract

The invention discloses a composition for improving fatty liver. The composition is prepared from 0.9-2.7 parts by mass of quinoa and 0.03-0.1 part by mass of lycium barbarum polysaccharide. The invention further discloses application of the composition for improving the fatty liver in preparation of health care products and diets for improving the fatty liver. Fatty liver is non-alcoholic fatty liver. Experiments find that the quinoa and lycium barbarum polysaccharide diet significantly reduces lipid deposition in liver cells, improves lipid metabolism disorder, effectively relieves the burden of the liver in high-fat diet, improves the liver health in high-fat diet, can increase the content of short-chain fatty acids in a human body in a mode of simultaneous supplement of medicine and food, and improves the immunity of the human body. The fatty liver symptom is improved. Meanwhile, the product is high in safety to human bodies, does not generate side effects to the human bodies, and can be eaten trustingly.

Description

Technical Field

[0001] The present invention relates to the technical field of fatty liver, and in particular to the use of quinoa combined with wolfberry polysaccharide diet in the preparation of a method for promoting the production of short-chain fatty acids in the intestine and improving fatty liver. Background Art

[0002] Nonalcoholic fatty liver disease refers to a clinical pathological syndrome characterized by excessive fat deposition in hepatocytes caused by factors other than alcohol and other clear liver-damaging factors. It is a multisystem disease associated with obesity, type 2 diabetes, and cardiovascular disease. The global prevalence of nonalcoholic fatty liver disease is as high as 20% to 33%, and it has become an important public health issue that seriously threatens human health. Dietary intervention and regulation of intestinal microecology play a key role in the prevention and treatment of nonalcoholic fatty liver disease.

[0003] Lycium barbarum polysaccharides, as one of the main active ingredients of wolfberry, exhibit a wide range of biological properties. Studies have shown that Lycium barbarum polysaccharides can promote the growth of probiotics in vitro and regulate intestinal microbiota, showing the potential of probiotics. Some studies have found that combining Lycium barbarum polysaccharides with aerobic exercise to treat non-alcoholic fatty liver disease is more effective in improving liver damage and reducing lipid and inflammation levels than administering Lycium barbarum polysaccharides alone. However, aerobic exercise is a lifestyle intervention that has a good weight loss effect on patients with non-alcoholic fatty liver disease, but it is usually difficult to follow it for a long time. Therefore, it is necessary to find natural foods that are compatible with Lycium barbarum polysaccharides to improve fatty liver. For this reason, we proposed the use of quinoa combined with Lycium barbarum polysaccharides in the preparation of a diet to improve fatty liver. Summary of the invention

[0004] The present application provides a composition for improving fatty liver and its application, which improves and alleviates the problem of non-alcoholic fatty liver.

[0005] The present application provides a composition for improving fatty liver, comprising: 0.9-2.7 parts by mass of quinoa and 0.03-0.1 parts by mass of wolfberry polysaccharide.

[0006] Preferably, it comprises: 1.8 parts by mass of quinoa and 0.05 parts by mass of wolfberry polysaccharide.

[0007] Preferably, the quinoa is 1.8 g and the wolfberry polysaccharide is 50 mg.

[0008] Preferably, the quinoa is combined with wolfberry polysaccharide to promote the production of short-chain fatty acids.

[0009] Preferably, the quinoa preparation method comprises the following steps:

[0010] Roast the washed and dried quinoa at 52°C;

[0011] The fried quinoa is crushed by low-temperature vibration in a vibrating superfine grinder;

[0012] The crushed quinoa was filtered through a 300-mesh sieve.

[0013] Preferably, the composition comprises capsules, pills, emulsions, tablets, powders.

[0014] The invention also includes the use of a composition for improving fatty liver in preparing health products and diets for improving fatty liver.

[0015] Preferably, the fatty liver is non-alcoholic fatty liver.

[0016] It can be seen from the above technical solutions that the present application provides a composition for improving fatty liver and its application.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] 1. This application can be used as a way of combining medicine and food to relieve the pathological changes of fatty liver under a high-fat diet. At the same time, this application is highly safe for the human body, has no side effects on the body, and can be consumed with confidence;

[0019] 2. This application can be taken for a long time, which can not only effectively prevent fatty liver, but also increase the content of short-chain fatty acids in the intestine and improve the health of the body;

[0020] 3. This application maximizes the combined effect of quinoa and wolfberry polysaccharides by processing quinoa in the optimal ratio, significantly reduces the fat content in the liver during a high-fat diet, and effectively alleviates the occurrence of fatty liver.

[0021] In summary, the present application discovered through experiments that a quinoa combined with wolfberry polysaccharide diet significantly reduced lipid accumulation in liver cells and improved lipid metabolism disorders, effectively alleviated the burden on the liver under a high-fat diet, and improved liver health under a high-fat diet. It can promote the production of short-chain fatty acids and improve fatty liver symptoms through the combination of medicine and food, and also provide important nutrients for the human body. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solution of the present application, the following is a brief introduction to the drawings required for use in the implementation cases. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0023] Figure 1 To investigate the effect of quinoa combined with wolfberry polysaccharide diet on energy intake in rats with nonalcoholic fatty liver disease.

[0024] Figure 2The changes in body weight of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0025] Figure 3 The changes of Lee's index in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0026] Figure 4 To investigate the changes of liver indexes in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0027] Figure 5 To investigate the changes in visceral fat index in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides.

[0028] Figure 6 Pathological staining of liver in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa and wolfberry polysaccharides (200×);

[0029] Figure 7 To investigate the changes in TC levels in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0030] Figure 8 To investigate the changes in TG levels in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0031] Fig. 9 To investigate the changes in AST levels in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0032] Fig.10 To investigate the changes of ALT levels in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0033] Fig.11 To investigate the changes in LDL-C levels in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0034] Fig.12 To investigate the changes in HDL-C levels in rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0035] Fig.13 To investigate the changes of TC levels in liver tissue of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0036] Fig.14 To investigate the changes of TG levels in liver tissue of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0037] Fig.15 To study the changes of FFA levels in liver tissue of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides;

[0038] Fig.16 To study the changes of acetic acid content in the colon of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides.

[0039] Fig.17 To study the changes of propionic acid content in colon of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides.

[0040] Fig.18 To study the changes of butyrate content in the colon of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides.

[0041] Fig.19 To study the changes of isobutyric acid content in colon of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa and wolfberry polysaccharides.

[0042] Fig. 20 To study the changes of isovaleric acid content in the colon of rats with non-alcoholic fatty liver disease after dietary intervention with quinoa and wolfberry polysaccharides.

[0043] Fig.21 To investigate the changes in valeric acid content in the colon of rats with nonalcoholic fatty liver disease after dietary intervention with quinoa combined with wolfberry polysaccharides. DETAILED DESCRIPTION

[0044] In order to enable those skilled in the art to better understand the technical solutions in the present application, the technical solutions in the embodiments of the present application will be clearly and completely described below in conjunction with the accompanying drawings;

[0045] In this application, the composition includes capsules, pills, emulsions, tablets, and powders.

[0046] In the present application, the composition for improving fatty liver is used in the preparation of health products and diets for improving fatty liver.

[0047] In the application, fatty liver refers to non-alcoholic fatty liver.

[0048] In a specific embodiment, the present application conducts a study on the effect of quinoa combined with wolfberry polysaccharide diet in alleviating fatty liver disease, and through the therapeutic effect of quinoa combined with wolfberry polysaccharide diet on rats with non-alcoholic fatty liver disease induced by a high-fat diet and the effect on regulating short-chain fatty acids involved in anti-non-alcoholic fatty liver disease, it is used to confirm that quinoa combined with wolfberry polysaccharide diet significantly reduces lipid accumulation in hepatocytes and improves lipid metabolism disorders, thereby achieving the purpose of improving fatty liver.

[0049] It is worth noting that the dosage in this application is the dosage for experimental rats, and the specific dosage for humans can be calculated by referring to the existing technology (refer to the table of "equivalent dose ratios between humans and animals calculated by body surface area" in "Pharmacological Experimental Methodology" edited by Professor Xu Shuyun to calculate the dosage of the drug used).

[0050] Sources and preparation of Lycium barbarum polysaccharides and quinoa in this application

[0051] Lycium barbarum polysaccharides were provided by Ningxia Bairuiyuan Lycium barbarum Co., Ltd.;

[0052] Quinoa was provided by Songwa Village, Ningxia. The preparation method comprises the following steps:

[0053] Fry the washed and dried quinoa at 52°C for more than 10 minutes;

[0054] The fried quinoa is crushed by low-temperature vibration in a vibrating superfine grinder;

[0055] Filter the crushed quinoa through a 300-mesh sieve.

[0056] In this application;

[0057] Quinoa nutritional composition analysis

[0058] The crude protein, fat, ash and dietary fiber contents were determined by the Kjeldahl method (GB5009.5-2016), Soxhlet extraction method (GB5009.6-2016), dry ash method (GB5009.4-2016) and gravimetric method (GB5009.88-2014) of the Chinese Food Safety National Standard (GB). The starch and amylose contents were determined by kits (A148-1-1, A152-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The resistant starch and non-resistant starch contents were determined by kits (ADS-W-DF009-48, Jiangsu Aidi Biotechnology Co., Ltd., Jiangsu, China).

[0059] Animal experiments

[0060] During the experiment, all animal operation protocols were approved by the Experimental Animal Ethics Committee of Ningxia Medical University (IACUC-NYLAC-2020-263).

[0061] Example 1

[0062] Male SD rats (180-220 g) were obtained from the Experimental Animal Center of Ningxia Medical University and were raised under specific conditions (temperature: 20-25°C, humidity: 45-55%, 12 h light / dark cycle). After the adaptation period, they were randomly divided into two groups: a normal group (NC group, n = 6, normal feed provided by Beijing Keao Xieli Feed Co., Ltd.) and a model group (n = 24, fed with a high-fat diet containing 60% fat, MD12033, Medison Biopharmaceutical Co., Ltd.).

[0063] Twelve weeks after modeling, the rats in the model group were randomly divided into the following four groups:

[0064] (1) Model group (MC group, n=6): fed with a high-fat diet (60 kcal% fat).

[0065] (2) Quinoa group (QC group, n=6): The mice were fed with a high-fat diet containing quinoa superfine powder.

[0066] (3) Lycium barbarum polysaccharide group (LC group, n=6): mice were fed a high-fat diet containing Lycium barbarum polysaccharide.

[0067] (4) Quinoa combined with Lycium barbarum polysaccharide group (QL group, n = 6): The mice were fed with a high-fat diet containing both quinoa superfine powder and Lycium barbarum polysaccharide.

[0068] All rats had free access to food and water during the experiment. Food intake and body weight were measured and recorded weekly. At 11 weeks of intervention, rats were fasted for 12 h and then euthanized under ether anesthesia. The collected blood samples were centrifuged at 2000 × g and 4 °C for 10 min to obtain serum. Liver, epididymal, and perirenal adipose tissue samples were collected, and the tissue samples were quickly frozen in liquid nitrogen and stored at −80 °C for further analysis. The dietary composition is summarized in Table 1.

[0069] Table 1. High-fat diet and intervention diet formula (mass percentage %)

[0070]

[0071] Histological analysis

[0072] Liver tissues were fixed in 10% neutral formalin and embedded in paraffin for histological analysis of pathological changes. Sections (thickness 4 μm) were stained with hematoxylin-eosin (H&E) and Oil Red O and then observed using an optical microscope (Nikon, Japan). The histology of liver damage was evaluated using the nonalcoholic fatty liver disease activity score (NAS) (Kleiner et al., 2005).

[0073] Biochemical and ELISA tests

[0074] Plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), liver total cholesterol (liver TC), liver triglyceride (liver TG), and free fatty acid (FFA) in liver tissue were detected using kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Liver tissue homogenate was prepared by homogenizing 0.1 g of liver tissue with 0.9 mL of phosphate buffer in an ultrasonic tissue grinder, and the supernatant was used for analysis.

[0075] Determination of short-chain fatty acids (SCFAs) content

[0076] Colon samples (approximately 50 mg) were homogenized with 100 mg of glass beads in 500 μL of water for 1 min and then centrifuged at 12,000 rpm / min for 10 min at 4°C. Subsequently, 200 μL of the supernatant and 20 μL of a 375 μL / mL 4-methylvaleric acid solution as an internal standard mixture were continuously stirred, acidified with 100 μL of 15% phosphoric acid, and the resulting solution was extracted with 280 μL of ether and stirred continuously for 1 min. Then, centrifuged at 4°C, 12,000 rpm for 10 min. The supernatant was transferred to a vial and analyzed using a gas chromatograph-mass spectrometer. SCFAs were identified by comparing their mass spectrometry results with the retention times of six standard SCFAs (acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid).

[0077] Statistical methods

[0078] SPSS24.0 software was used for statistical analysis of all data. Data were expressed as mean ± standard error. When variance was equal, variance analysis was used for comparison between multiple samples. Tukey-Kramer and LSD were used for post hoc comparison between groups. When variance was unequal, Kruskal-Wallis nonparametric test was used for comparison between multiple samples. Bonferroni method was used for post hoc comparison between two groups. The test level was α = 0.05. Graphpadprism 9.0 was used for drawing.

[0079] Example 2

[0080] Table 2 shows the nutritional content of 52℃ quinoa superfine powder

[0081] Table 2 Nutritional content of 52℃ quinoa superfine powder

[0082]

[0083] Example 3

[0084] Effect of quinoa combined with wolfberry polysaccharide diet on energy intake in rats with non-alcoholic fatty liver disease

[0085] This example studies the effect of quinoa combined with wolfberry polysaccharides as daily dietary sources on rats with non-alcoholic fatty liver disease, and conducts animal experiments. At the end of 12 weeks of modeling, the energy intake of each group was NC group (327.00±6.85kcal / w), MC group (294.83±6.85kcal / w), QC group (314.10±11.91kcal / w), LC group (295.75±6.24kcal / w), QL group (315.02±4.46kcal / w). There was no statistical difference in energy intake among the five groups, and P values ​​were all >0.05. At the end of 11 weeks of intervention, there was no statistical difference in energy intake among the five groups of rats, as shown in Figure 2. Figure 1 shown.

[0086] Quinoa combined with wolfberry polysaccharide diet reduces body weight in rats with non-alcoholic fatty liver disease

[0087] At the end of 12 weeks of modeling, the body weights of rats in each group were NC group (520.48±13.98g), MC group (650.98±19.62g), QC group (649.53±17.08g), LC group (632.95±13.64g), and QL group (630.48±17.27g). Compared with the NC group, the body weights of rats in the MC group, QC group, LC group, and QL group increased significantly, and there were statistical differences (all P values ​​were <0.01). Compared with the MC group, there were no statistically significant differences in the body weights of rats in the QC, LC, and QL groups (all P values ​​were >0.05).

[0088] At the end of the experiment,

[0089] In terms of body weight, compared with the NC group, the final body weight of rats in the MC group increased significantly, and the difference between the two groups was statistically significant (P < 0.001). Compared with the MC group, the final body weight values ​​of rats in the QC group, LC group, and QL group were all lower, with the most significant decrease in the QL group (P < 0.001). Compared with the final body weight values ​​of rats in the QL group, the body weight values ​​of rats in the QC group and LC group were higher, and the difference between the QC group and the QL group was statistically significant (P < 0.05). At the end of modeling, the body weight and Lee's index of rats in the MC, QC, LC, and QL groups were significantly higher than those in the NC group (P < 0.05). At the end of the intervention, the Lee's index of rats in the MC group was significantly higher than that in the NC group, QC group, and QL group, and the P values ​​were all < 0.05. There was no statistically significant difference between the LC group and the MC group (P > 0.05). Among all intervention groups, the QL intervention effect was better as shown in Table 3, Figure 2 and Figure 3 shown.

[0090] Table 3 Changes in body weight of rats with non-alcoholic fatty liver disease after intervention with quinoa combined with wolfberry polysaccharides (g)

[0091]

[0092] Note: *Compared with MC group, * P<0.05, ** P<0.01, *** P<0.001, ^Compared with the QL group, ^ P<0.05.

[0093] The liver index and visceral fat index of the MC group were higher than those of the other groups. Compared with the MC group, the liver index of the QC group and the QL group was significantly different (P QC <0.05, P QL <0.001)( Figure 4 In addition, the MC group had a higher visceral fat index than the other groups, and QL intervention reduced liver index and visceral fat index more significantly than LC or QC intervention alone ( Figure 5 ). The above results suggest that QL can reduce Lee's index, liver index and visceral fat index in rats with non-alcoholic fatty liver disease, and has a better fat-reducing effect than QC and LC, and is independent of calorie intake, such as Figure 4 and Figure 5 shown.

[0094] Example 4

[0095] Effects of quinoa combined with wolfberry polysaccharide intervention on liver pathology and lipid markers in rats with non-alcoholic fatty liver disease

[0096] Nonalcoholic fatty liver disease affects the morphological structure of liver tissue and lipid metabolism disorders. Histopathology showed that there were large areas of lipid droplets of different sizes in the liver tissue of the MC group. Compared with the MC group, the area of ​​lipid droplets in the liver tissue of the NC group, QC group, LC group and QL group after intervention was significantly reduced. Among them, the effect of QL intervention on improving lipid deposition was more obvious ( Figure 5 ). In addition, the results showed that compared with the MC group, the plasma TG, TC, LDL-C, AST and ALT concentrations in the NC group and the QL group were significantly lower. The plasma HDL-C concentration in the MC group was significantly lower than that in the other four groups ( Figure 6-Figure 15 ). Compared with the MC group, the levels of TG, TC, and FFA in liver tissue were significantly decreased in the NC group, QC group, LC group, and QL intervention group. These data indicate that QL intervention improved liver damage and liver function and reduced lipid deposition in the liver. Figure 5-14 shown.

[0097] Example 5

[0098] Quinoa combined with Lycium barbarum polysaccharides promotes the production of short-chain fatty acids

[0099] Compared with the NC group, the acetic acid level in the MC group decreased, and the difference was statistically significant (P<0.05). Compared with the MC group, the acetic acid level in the QC group, LC group, and QL group increased. There was no statistical difference in acetic acid level between the LC group and the MC group (P>0.05), and the differences between the QC group and the QL group and the MC group were statistically significant (P<0.05). Fig.16 ).

[0100] Compared with the NC group, the propionic acid level in the MC group decreased, and the difference was statistically significant (P<0.05). Compared with the MC group, the propionic acid levels in the QC group, LC group, and QL group increased. There was no statistical difference in propionic acid between the LC group and the MC group (P>0.05), and there were statistically significant differences between the QC group and the QL group and the MC group (P<0.01). Fig.17 ).

[0101] Compared with the NC group, the butyric acid level in the MC group was lower, and the difference was statistically significant (P<0.05). Compared with the MC group, the butyric acid level in the QC group, LC group, and QL group increased, and the difference in butyric acid concentration between the QL group and the MC group was statistically significant (P<0.05). Fig.18 ).

[0102] Compared with the MC group, the isobutyric acid levels in the NC group, QC group, LC group, and QL group increased, but the differences were not statistically significant (P<0.05). Fig.19 ).

[0103] Compared with the NC group, the isovaleric acid level in the MC group decreased, with no significant difference (P>0.05). Compared with the MC group, the isovaleric acid level in the QC and QL groups increased, while the isovaleric acid level in the LC group decreased, with no significant difference (P>0.05).

[0104] ( Fig. 20 ).

[0105] Compared with the NC group, the valeric acid level in the MC group decreased, and the difference was statistically significant (P < 0.05). Compared with the MC group, the valeric acid level in the QC group, LC group, and QL group increased, and the difference was not statistically significant (P > 0.05). Fig.21 ).

[0106] In this example, a rat model of non-alcoholic fatty liver disease was established using a high-fat diet, and dietary intervention with quinoa and wolfberry polysaccharides protected rats from the effects of a high-fat diet on non-alcoholic fatty liver disease, further improved intestinal and hepatic lipid metabolism disorders, promoted the production of short-chain fatty acids in the colon, and effectively protected the intestinal liver. These findings suggest that QL has potential value as a beneficial dietary therapeutic agent in non-alcoholic fatty liver disease.

[0107] The research findings of this application show that the combination of 1.8g quinoa and 50mg Lycium barbarum polysaccharides increases the level of short-chain fatty acids in the intestine, especially butyrate production, improves lipid metabolism disorders, reduces visceral fat, and effectively alleviates the progression of non-alcoholic fatty liver disease. Therefore, the combination of 1.8g quinoa and 50mg Lycium barbarum polysaccharides becomes a promising new dietary pattern for improving non-alcoholic fatty liver disease.

[0108] Those skilled in the art will readily appreciate other embodiments of the present application after considering the specification and practicing the application disclosed herein. The present application is intended to cover any variations, uses or adaptations of the present application, which follow the general principles of the present application and include common knowledge or customary techniques in the art that are not disclosed in the present application. The specification and examples are intended to be exemplary only, and the true scope of the present application is indicated by the claims.

[0109] It should be understood that the present application is not limited to the precise structure described above and shown in the drawings, and various modifications and changes can be made without departing from its scope. The above-described embodiments of the present application do not constitute a limitation on the scope of protection of the present application.

Claims

1. A composition for improving fatty liver, characterized in that: include: 0.9-2.7 parts by mass of quinoa and 0.03-0.1 parts by mass of wolfberry polysaccharide.

2. A composition for improving fatty liver according to claim 1, characterized in that: include: 1.8 parts by mass of quinoa and 0.05 parts by mass of wolfberry polysaccharides.

3. A composition for improving fatty liver according to claim 1, characterized in that: The quinoa combined with the wolfberry polysaccharide diet promotes the generation of short-chain fatty acids.

4. A composition for improving fatty liver according to claim 1, characterized in that: The quinoa preparation method comprises the following steps: Roast the washed and dried quinoa at 52℃-53℃; The fried quinoa is crushed by low-temperature vibration in a vibrating superfine grinder; The crushed quinoa was filtered through a 300-mesh sieve.

5. The composition for improving fatty liver according to claim 1, characterized in that: The compositions include capsules, pills, emulsions, tablets, and powders.

6. Use of the composition for improving fatty liver according to any one of claims 1 to 5 in preparing health products and diets for improving fatty liver.

7. The use according to claim 6, characterized in that: The fatty liver is non-alcoholic fatty liver.

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