An anti-inflammatory and fat-reducing ferment and a method for preparing the same

Abstract

This invention belongs to the field of microbial fermentation technology, specifically relating to an anti-inflammatory and fat-reducing fermented product and its preparation method. The anti-inflammatory and fat-reducing fermented product comprises, by weight, the following raw materials: 150-300 parts purslane, 20-80 parts perilla leaves, 10-40 parts sunflower flowers, 1-10 parts golden camellia, 0.15-3 parts fermentation starter culture powder, and 6-30 parts enzyme preparation. This patent studies the relationship between obesity and inflammation from an inflammatory perspective, providing a fermented product with anti-inflammatory and fat-reducing effects. This fermented product is rich in functions and has good long-term effects.

Description

Technical Field

[0001] This invention belongs to the field of microbial fermentation technology, specifically relating to an anti-inflammatory and fat-reducing ferment and its preparation method. Background Technology

[0002] Many inflammatory conditions in the body, such as chronic gastritis, cholecystitis, and skin inflammation, produce inflammatory factors. These factors circulate throughout the body via the bloodstream, leading to systemic chronic inflammation. If this systemic chronic inflammation is not treated thoroughly, it can cause metabolic disorders, such as insulin resistance, and consequently, obesity. Furthermore, inflammatory factors primarily attack fat cells, causing a sustained increase in inflammatory factor levels in fat cells, exacerbating inflammation and obesity. Therefore, obese people are more prone to weight gain because even without excessive energy intake, their fat cells are disrupted by the attack of inflammatory factors, resulting in long-term obesity that is difficult to improve. The WHO defines obesity as a body mass index (BMI) greater than or equal to 30. Obesity can induce hypertension, type 2 diabetes, cardiovascular disease, osteoarthritis, kidney failure, liver disease, asthma, depression, and various cancers.

[0003] Currently, there are four main approaches to treating obesity: physical exercise, dietary intervention, drug therapy, and surgery. Physical exercise has always been a popular and accepted method for weight loss. By systematically increasing energy expenditure and reducing fat content, it is considered safe and effective, but it's crucial to employ scientific exercise training. Dietary intervention, or energy intake control, requires strict control of the proportions of sugar, fat, and protein in the diet, as well as balancing macro- and micro-elements. It is generally believed that reducing daily intake by about 25% is sufficient to control energy intake and achieve weight loss. Drug therapy is essentially similar to energy intake control, and it is divided into appetite suppressants and digestive blockers. Orlistat has shown positive results in weight loss trials and is an over-the-counter drug in the United States, primarily used to reduce individual energy intake for weight loss. Surgery is more effective for severely obese patients. However, physical exercise is difficult to maintain long-term, and ordinary people cannot guarantee scientific exercise training, which can easily lead to physical injury. Dietary intervention goes against a person's appetite, and achieving adequate supplementation of various nutrients is also difficult. Drug therapy has significant side effects. Surgery, on the other hand, is more harmful to the body and is mainly for severely obese patients.

[0004] Functional foods refer to foods with specific nutritional and health benefits, meaning they are suitable for specific groups of people, regulate bodily functions, and are not intended for treatment. For example, Chinese patent document CN114949039A discloses a plant ferment and its use in preparing a weight-loss composition. The ferment is made by mixing mulberry (Mours alba), pomegranate, purslane, bitter melon, and fennel in a ratio of 0.5-4:4-8:0.5-4:0.1-2:0.5-4, obtaining plant extracts through solvent extraction, and then fermenting the mixture. However, this patent separates inflammation and weight loss, resulting in a single product. Summary of the Invention

[0005] Therefore, this patent studies the relationship between obesity and inflammation from the perspective of inflammation, and provides a fermented product with anti-inflammatory and fat-reducing effects. This fermented product has rich functions and good long-term effects.

[0006] The technical solution of this invention is as follows:

[0007] On the one hand, the present invention provides an anti-inflammatory and fat-reducing fermented product, wherein the fermentation raw materials include, by weight, 150-300 parts of purslane, 20-80 parts of perilla leaves, 10-40 parts of sunflower flowers, 1-10 parts of golden camellia, 0.15-3 parts of fermentation starter powder and 6-30 parts of enzyme preparation.

[0008] Preferably, the fermentation raw materials include, by weight: 150-200 parts purslane, 20-60 parts perilla leaves, 10-30 parts sunflower flowers, 1-5 parts golden camellia, 0.15-2 parts fermentation starter, and 6-20 parts enzyme preparation.

[0009] Preferably, the fermentation powder includes *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium adolescentis*, with the weight ratio of each strain being 0.05-0.9:0.09-0.5:0.2-0.6.

[0010] Preferably, the enzyme preparation includes cellulase, pectinase and acidic protease, with the weight ratio of each enzyme being 2-10:2-10:2-10.

[0011] More preferably, the viable count of *Lactobacillus plantarum* is 100-300 billion CFU / g, the viable count of *Lactobacillus acidophilus* is 100-500 billion CFU / g, and the viable count of *Bifidobacterium adolescentis* is 100-300 billion CFU / g.

[0012] More preferably, the cellulase has an activity of 10,000-30,000 U / g, the pectinase has an activity of 10,000-50,000 U / g, and the acidic protease has an activity of 50,000-100,000 U / g.

[0013] More preferably, the present invention provides an anti-inflammatory and fat-reducing fermented product, comprising 200 parts of purslane, 50 parts of perilla leaves, 20 parts of sunflower flowers, 5 parts of golden camellia, 0.3 parts of *Lactobacillus plantarum*, 0.3 parts of *Lactobacillus acidophilus*, 0.4 parts of *Bifidobacterium adolescentis*, 5 parts of cellulase, 5 parts of pectinase, and 5 parts of acidic protease.

[0014] Specifically, the cellulase activity is 20,000 U / g, the pectinase activity is 30,000 U / g, and the acidic protease activity is 100,000 U / g; the viable counts of *Lactobacillus plantarum* are 200 billion CFU / g, *Lactobacillus acidophilus* are 200 billion CFU / g, and *Bifidobacterium adolescentis* are 200 billion CFU / g.

[0015] On the other hand, the present invention provides a method for preparing an anti-inflammatory and fat-reducing ferment, comprising the following steps: (1) pulverizing purslane, perilla leaves, sunflower flowers and golden camellia into powder and sieving to a mesh size of 100-600 mesh, adding water and mixing evenly, and adjusting the pH to 4.0-5.0 to obtain a mixed solution; (2) adding an enzyme preparation to the mixed solution for enzymatic hydrolysis, and adjusting the pH to 5.0-6.0 after the enzymatic hydrolysis is completed to obtain an enzymatic hydrolysate; (3) extracting the enzymatic hydrolysate at a temperature of 80-102°C for 10-90 min to obtain an extract; (4) adding fermentation bacteria powder to the extract for fermentation to obtain a fermentation broth, and separating the solid and liquid of the fermentation broth to obtain a fermentation clear liquid, which is the fermented product.

[0016] Preferably, the enzymatic hydrolysis conditions in step (2) are a temperature of 40-70°C and a time of 90-240 min; the fermentation conditions in step (4) are a temperature of 15-45°C and a time of 36-96 h.

[0017] More preferably, anhydrous citric acid is used to adjust the pH in step (1); sodium bicarbonate is used to adjust the pH in step (2).

[0018] The technical solution of this invention has the following advantages:

[0019] 1. The anti-inflammatory and fat-reducing fermented raw material provided by this invention contains purslane, whose main chemical components include flavonoids, coumarins, terpenes, steroids, alkaloids, amino acids, various pigments, and minerals. Among them, alkaloids, flavonoids, and steroids all have good anti-inflammatory activity.

[0020] The significant anti-inflammatory activity of perilla leaves has made them a new focus of research. Their abundant volatile oil components, flavonoids, and phenolic acids are the main material basis for this anti-inflammatory activity. Identified active ingredients include perillaldehyde, isoperillone, luteolin, and rosmarinic acid.

[0021] Tea polyphenols and flavonoids are considered the main active ingredients in tea, while Camellia species contain relatively few triterpenoid saponins. Golden camellia contains a large amount of triterpenoids, which are the main functional substances responsible for its anti-inflammatory activity.

[0022] The four components—purslane, perilla leaves, golden camellia, and sunflower flowers—work together through enzymatic fermentation to achieve excellent anti-inflammatory and fat-reducing effects. They reduce fat from an anti-inflammatory perspective, truly adjusting metabolic function from within the body, restoring normal fat metabolism, and providing long-lasting results. Once the body's metabolism is regulated, obesity may even remain permanent.

[0023] 2. The anti-inflammatory and fat-reducing fermented composition provided by the present invention, by selecting appropriate proportions of raw materials and fermentation powder, can further reduce the content of inflammatory factors in rat serum, weight gain and body fat percentage, thereby achieving better anti-inflammatory and fat-reducing effects.

[0024] 3. The anti-inflammatory and fat-reducing fermentation composition provided by the present invention can adjust the pH value during the fermentation process in a timely manner, making it more suitable for the production of the strain and improving the fermentation yield.

[0025] 4. The medicinal materials used in this invention are all safe food ingredients that can be used for both medicinal and edible purposes. They are made through fermentation and processing of traditional Chinese medicine and are safe and reliable. Attached Figure Description

[0026] Figure 1 Box plot of initial body weight distribution in rats

[0027] Figure 2 Normal PP plot of initial body weight of rats Detailed Implementation

[0028] To make the objectives, technical solutions, and advantages of this invention clearer, the embodiments of this invention will be described in further detail below. All other embodiments obtained by those skilled in the art based on the embodiments disclosed in this invention without inventive effort are within the scope of protection of this disclosure.

[0029] For any experimental steps or conditions not specified in the examples, the procedures or conditions described in the literature in this field can be followed. All reagents without specified manufacturers are commercially available standard reagent products.

[0030] In the following examples, the source information for some products is as follows:

[0031] Purslane is the dried aerial part of the purslane plant, belonging to the Portulacaceae family;

[0032] Perilla leaves are the dried leaves of the perilla plant, belonging to the Lamiaceae family.

[0033] Sunflower pollen is made from sunflower flowers, specifically from bee pollen or broken-cell wall pollen.

[0034] Golden camellia is the edible part of the camellia plant, belonging to the Theaceae family and the Camellia genus.

[0035] Lactobacillus plantarum is Lactobacillus plantarum LP-A3 from Shandong Zhongke Jiayi Bioengineering Co., Ltd.;

[0036] Lactobacillus acidophilus is Lactobacillus acidophilus LA-10A from Sichuan Gaofuji Biotechnology Co., Ltd.;

[0037] Bifidobacterium adolescentis was purchased from Jiangsu Xinshengao Biotechnology Co., Ltd.

[0038] Example 1

[0039] This embodiment provides an anti-inflammatory and fat-reducing fermented product, comprising 200g of purslane, 50g of perilla leaves, 20g of sunflower flowers, 10g of golden camellia, 0.3g of *Lactobacillus plantarum*, 0.3g of *Lactobacillus acidophilus*, 0.4g of *Bifidobacterium adolescentis*, 5g of cellulase, 5g of pectinase, 5g of acidic protease, and 3720g of water.

[0040] Its preparation method is as follows:

[0041] (1) 200g of purslane, 50g of perilla leaves, 20g of sunflower flowers and 10g of golden camellia are crushed and passed through a 300-mesh sieve. They are then mixed with 3720g of water and 0.35g of anhydrous citric acid is added to adjust the pH to 4.8 to obtain a mixed solution.

[0042] (2) Add 5g of cellulase, 5g of pectinase and 5g of acidic protease to the mixture for enzymatic hydrolysis. Hydrolyze at 58°C for 150min. After the enzymatic hydrolysis is completed, add 3g of sodium bicarbonate to adjust the pH to 5.6 to obtain the enzymatic hydrolysate.

[0043] The cellulase activity was 20,000 U / g, the pectinase activity was 30,000 U / g, and the acidic protease activity was 100,000 U / g.

[0044] (3) Heat the enzymatic hydrolysate to 95°C and stir for 30 min to obtain the extract;

[0045] (4) When the temperature drops to 35°C, add 0.3g of Lactobacillus plantarum, 0.3g of Lactobacillus acidophilus and 0.4g of Bifidobacterium adolescentis to the extract, and ferment at 35°C for 72 hours to obtain the fermentation liquid;

[0046] The viable counts of *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium adolescentis* were 200 billion CFU / g.

[0047] (5) Separate the solid and liquid of the above fermentation broth to obtain the fermentation clear broth, i.e., the fermentation product. The pH of the fermentation clear broth is 3.7, the soluble solids content is 10.2%, and the total acid is 1.76% (calculated as lactic acid).

[0048] Example 2

[0049] This embodiment provides an anti-inflammatory and fat-reducing fermented product, comprising 150g of purslane, 80g of perilla leaves, 30g of sunflower flowers, 8g of golden camellia, 0.05g of *Lactobacillus plantarum*, 0.35g of *Lactobacillus acidophilus*, 0.6g of *Bifidobacterium adolescentis*, 2g of cellulase, 10g of pectinase, 2g of acidic protease, and 4500g of water.

[0050] Its preparation method is as follows.

[0051] (1) 150g of purslane, 80g of perilla leaves, 30g of sunflower flowers and 8g of golden camellia are crushed and passed through a 300-mesh sieve. They are then mixed with 4500g of water and 0.48g of anhydrous citric acid is added to adjust the pH to 4.8 to obtain a mixed solution.

[0052] (2) Add 2g of cellulase, 10g of pectinase and 2g of acidic protease to the mixture for enzymatic hydrolysis. Hydrolyze at 58°C for 150min. After hydrolysis, add 3.7g of sodium bicarbonate to adjust the pH to 5.6 to obtain the hydrolysate.

[0053] The cellulase activity was 20,000 U / g, the pectinase activity was 30,000 U / g, and the acidic protease activity was 100,000 U / g.

[0054] (3) Heat the enzymatic hydrolysate to 95°C and stir for 30 min to obtain the extract;

[0055] (4) When the temperature drops to 35°C, add 0.05g of Lactobacillus plantarum, 0.35g of Lactobacillus acidophilus and 0.6g of Bifidobacterium adolescentis to the extract, and ferment at 35°C for 72 hours to obtain the fermentation liquid;

[0056] The viable counts of Lactobacillus plantarum were 200 billion CFU / g, Lactobacillus acidophilus were 200 billion CFU / g, and Bifidobacterium adolescentis were 200 billion CFU / g.

[0057] (5) Separate the solid and liquid of the above fermentation broth to obtain the fermentation clear broth, i.e., the fermentation product. The pH of the fermentation clear broth is 3.9, the soluble solids content is 8.1%, and the total acid is 1.52% (calculated as lactic acid).

[0058] Example 3

[0059] This embodiment provides an anti-inflammatory and fat-reducing fermented product, comprising 220g of purslane, 57g of perilla leaves, 23g of sunflower flowers, 3g of golden camellia, 0.6g of *Lactobacillus plantarum*, 0.09g of *Lactobacillus acidophilus*, 0.31g of *Bifidobacterium adolescentis*, 4g of cellulase, 2g of pectinase, 10g of acidic protease, and 3950g of water.

[0060] Its preparation method is as follows

[0061] (1) 220g of purslane, 57g of perilla leaves, 23g of sunflower flowers and 3g of golden camellia are crushed and passed through a 300-mesh sieve. They are then mixed with 3950g of water and 0.4g of anhydrous citric acid is added to adjust the pH to 4.8 to obtain a mixed solution.

[0062] (2) Add 4g of cellulase, 2g of pectinase and 10g of acidic protease to the mixture for enzymatic hydrolysis. Hydrolyze at 58°C for 150min. After the hydrolysis is completed, add 4.2g of sodium bicarbonate to adjust the pH to 5.6 to obtain the hydrolysate.

[0063] The cellulase activity was 20,000 U / g, the pectinase activity was 30,000 U / g, and the acidic protease activity was 100,000 U / g.

[0064] (3) Heat the enzymatic hydrolysate to 95°C and stir for 30 min to obtain the extract.

[0065] (4) When the temperature drops to 35°C, add 0.6g of Lactobacillus plantarum, 0.09g of Lactobacillus acidophilus and 0.31g of Bifidobacterium adolescentis to the extract, and ferment at 35°C for 72 hours to obtain the fermentation liquid;

[0066] The viable counts of *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium adolescentis* were 200 billion CFU / g.

[0067] (5) Separate the solid and liquid of the above fermentation broth to obtain the fermentation clear broth, i.e., the fermentation product. The pH of the fermentation clear broth is 4.2, the soluble solids content is 10.3%, and the total acid is 1.77% (calculated as lactic acid).

[0068] Example 4

[0069] This embodiment provides an anti-inflammatory and fat-reducing fermented product, comprising 170g of purslane, 31g of perilla leaves, 35g of sunflower flowers, 6g of golden camellia, 0.4g of *Lactobacillus plantarum*, 0.1g of *Lactobacillus acidophilus*, 0.5g of *Bifidobacterium adolescentis*, 7g of cellulase, 6g of pectinase, 3g of acidic protease, and 4160g of water.

[0070] Its preparation method is as follows:

[0071] (1) 170g of purslane, 31g of perilla leaves, 35g of sunflower flowers and 6g of golden camellia are crushed and passed through a 300-mesh sieve. They are then mixed with 4160g of water and 0.54g of anhydrous citric acid is added to adjust the pH to 4.8 to obtain a mixed solution.

[0072] (2) Add 7g of cellulase, 6g of pectinase and 3g of acidic protease to the mixture for enzymatic hydrolysis. Hydrolyze at 58°C for 150min. After hydrolysis, add 4.68g of sodium bicarbonate to adjust the pH to 5.6 to obtain the hydrolysate.

[0073] The cellulase activity was 20,000 U / g, the pectinase activity was 30,000 U / g, and the acidic protease activity was 100,000 U / g.

[0074] (3) Heat the enzymatic hydrolysate to 95°C and stir for 30 min to obtain the extract;

[0075] (4) When the temperature drops to 35°C, add 0.4g of Lactobacillus plantarum, 0.1g of Lactobacillus acidophilus and 0.5g of Bifidobacterium adolescentis to the extract, and ferment at 35°C for 72 hours to obtain the fermentation liquid;

[0076] The viable counts of *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium adolescentis* were 200 billion CFU / g.

[0077] (5) Separate the solid and liquid of the above fermentation broth to obtain the fermentation clear broth, i.e., the fermentation product. The pH of the fermentation clear broth is 3.94, the soluble solids content is 11.2%, and the total acid is 1.83% (calculated as lactic acid).

[0078] Example 5

[0079] This embodiment provides an anti-inflammatory and fat-reducing fermented product, comprising 300g of purslane, 60g of perilla leaves, 10g of sunflower flowers, 5g of golden camellia, 0.3g of *Lactobacillus plantarum*, 0.5g of *Lactobacillus acidophilus*, 0.2g of *Bifidobacterium adolescentis*, 10g of cellulase, 2g of pectinase, 4g of acidic protease, and 4300g of water.

[0080] Its preparation method is as follows:

[0081] (1) 170g of purslane, 31g of perilla leaves, 35g of sunflower flowers and 6g of golden camellia are crushed and passed through a 300-mesh sieve. They are then mixed with 4300g of water and 0.54g of anhydrous citric acid is added to adjust the pH to 4.8 to obtain a mixed solution.

[0082] (2) Add 10g of cellulase, 2g of pectinase and 4g of acidic protease to the mixture for enzymatic hydrolysis. Hydrolyze at 58°C for 150min. After hydrolysis, add 4.68g of sodium bicarbonate to adjust the pH to 5.6 to obtain the hydrolysate.

[0083] The cellulase activity was 20,000 U / g, the pectinase activity was 30,000 U / g, and the acidic protease activity was 100,000 U / g.

[0084] (3) Heat the enzymatic hydrolysate to 95°C and stir for 30 min to obtain the extract;

[0085] (4) When the temperature drops to 35°C, add 0.3g of Lactobacillus plantarum, 0.5g of Lactobacillus acidophilus and 0.2g of Bifidobacterium adolescentis to the extract, and ferment at 35°C for 72 hours to obtain the fermentation liquid;

[0086] The viable counts of *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bifidobacterium adolescentis* were 200 billion CFU / g.

[0087] (5) Separate the solid and liquid of the above fermentation broth to obtain the fermentation clear broth, i.e., the fermentation product. The pH of the fermentation clear broth is 3.94, the soluble solids content is 11.2%, and the total acid is 1.83% (calculated as lactic acid).

[0088] Comparative Example 1

[0089] This comparative example provides an anti-inflammatory and fat-reducing fermented product, comprising 200g of purslane, 50g of perilla leaves, 20g of sunflower flowers, 10g of golden camellia, 0.35g of anhydrous citric acid, 3g of sodium bicarbonate, and 3720g of water.

[0090] Its preparation method is as follows:

[0091] (1) 200g of purslane, 50g of perilla leaves, 20g of sunflower flowers and 4g of golden camellia are crushed and passed through a 300-mesh sieve. They are then mixed with 3720g of water and 0.35g of anhydrous citric acid is added to adjust the pH to 4.8 to obtain a mixed solution.

[0092] (2) After keeping the above mixture at 58°C for 150 min, add 3g of sodium bicarbonate to adjust the pH to 5.6; (3) Heat the mixture obtained in step (3) to 95°C and stir for 30 min to obtain the extract;

[0093] (4) After cooling the extract to 35°C, keep it warm for 72 hours; then separate the solid and liquid to obtain the clear liquid.

[0094] The difference between this comparative example and Example 1 is that it did not undergo enzymatic hydrolysis and fermentation.

[0095] Comparative Example 2

[0096] This comparative example provides a fermented product that differs from Example 1 in that it does not include purslane. The weight ratio of sunflower flowers, perilla leaves, and golden camellia remains unchanged, and the total weight remains unchanged from the total weight of purslane, sunflower flowers, perilla leaves, and golden camellia in Example 1. Specifically, the amount of perilla leaves is 175g, sunflower flowers are 70g, and golden camellia is 35g.

[0097] The rest is the same as in Example 1.

[0098] Comparative Example 3

[0099] This comparative example provides a fermented product that differs from Example 1 in that it does not include perilla leaves. The weight ratio of sunflower flowers, purslane, and golden camellia remains unchanged, and the total weight is the same as that of purslane, sunflower flowers, perilla leaves, and golden camellia in Example 1. Specifically, the amount of purslane is 243.5g, sunflower flowers are 24.3g, and golden camellia is 12.2g.

[0100] The rest is the same as in Example 1.

[0101] Comparative Example 4

[0102] This comparative example provides a fermented product that differs from Example 1 in that it does not include sunflower flowers. The mass ratio of sunflower flowers, perilla leaves, and golden camellia remains unchanged, and the total weight remains unchanged from the total weight of purslane, sunflower flowers, perilla leaves, and golden camellia in Example 1. Specifically, the amount of purslane is 215.4g, perilla leaves are 58.3g, and golden camellia is 10.8g.

[0103] The rest is the same as in Example 1.

[0104] Comparative Example 5

[0105] This comparative example provides a fermented product that differs from Example 1 in that it does not include Camellia chrysantha. The mass ratio of sunflower flowers, perilla leaves and Camellia chrysantha remains unchanged, and the total weight remains unchanged from the total weight of purslane, sunflower flowers, perilla leaves and Camellia chrysantha in Example 1. Specifically, the purslane is 207.4g, the perilla leaves are 51.9g and the sunflower is 20.7g.

[0106] The rest is the same as in Example 1.

[0107] Comparative Example 6

[0108] This comparative example provides a fermentation product that differs from Example 1 in that the total bacterial strain mass of the fermentation powder remains unchanged, and it only includes 1g of Lactobacillus acidophilus. Everything else is the same as in Example 1.

[0109] Comparative Example 7

[0110] This comparative example provides a fermentation product that differs from Example 1 in that the total bacterial strain mass of the fermentation powder remains unchanged, and it only includes 1g of Bifidobacterium adolescentis. Everything else is the same as in Example 1.

[0111] Comparative Example 8

[0112] This comparative example provides a fermentation product that differs from Example 1 in that the total bacterial strain mass of the fermentation powder remains unchanged, and it only includes 1g of *Lactobacillus plantarum*. Everything else is the same as in Example 1.

[0113] Experiment Example 1: Rat Experiment

[0114] Test samples: The fermentation products of Examples 1-5 and Comparative Examples 1-8 were spray-dried and used as test samples.

[0115] Experimental Design:

[0116] SD rats, weighing 90-100g, male.

[0117] High-fat feed formula: 52.2% ordinary feed, 20% sucrose, 15% lard, 1.2% cholesterol, 0.2% sodium cholate, 10% casein, 0.6% calcium bicarbonate, 0.4% limestone powder, and 0.4% premix. Both ordinary and high-fat feeds were purchased from Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.

[0118] After a week of acclimatization feeding with a standard diet, rats were randomly divided into a normal control group (n=10) and a high-fat model group (n=140). The normal control group was fed a standard diet with a total energy content of 16.7 kJ / g, while the high-fat model group was fed a high-fat diet with a total energy content of 25.17 kJ / g. Ten rats were kept in each cage at a room temperature of 21℃-24℃ with a 12-hour day-night cycle of lighting. Rats had free access to water and food during the modeling period. The rats' weight and body length were measured the day before the experiment and at the end of each week. Daily weighing and recording of feed intake and uneaten feed were also recorded. After 8 weeks of modeling, except for the normal control group, rats in other groups exhibited obesity, greasy fur, dull and yellowish coat, loose stools, sluggish movement, lethargy, and significantly reduced activity, indicating successful modeling.

[0119] After successful modeling, 140 rats in the high-fat model group were randomly divided into the following groups: high-fat control group (model group), high-fat + Example 1 (Example 1 group), high-fat + Example 2 (Example 2 group), high-fat + Example 3 (Example 3 group), high-fat + Example 4 (Example 4 group), high-fat + Example 5 (Example 5 group), high-fat + Comparative Example 1 (Comparative Example 1 group), high-fat + Comparative Example 2 (Comparative Example 2 group), high-fat + Comparative Example 3 (Comparative Example 3 group), high-fat + Comparative Example 4 (Comparative Example 4 group), high-fat + Comparative Example 5 (Comparative Example 5 group), high-fat + Comparative Example 6 (Comparative Example 6 group), high-fat + Comparative Example 7 (Comparative Example 7 group), and high-fat + Comparative Example 8 (Comparative Example 8 group). All groups were fed a high-fat diet, but the Example and Comparative examples were administered the corresponding test sample by gavage at 0.9 g / kg, while the model group was administered the same volume of deionized water by gavage. All groups were fed this diet for 6 consecutive weeks.

[0120] At the end of week 6, rats were fasted for 12 hours but allowed free access to water and were weighed. The following morning, rats were anesthetized by intraperitoneal injection of 2% sodium pentobarbital solution at a dose of 50 mg / kg body weight. 10 ml of blood was then collected from the abdominal aorta and incubated at 4°C for 1 hour before centrifugation at 3500 rpm for 15 minutes. The serum was collected and stored at -20°C for ELISA testing. Adipose tissue from the abdomen, epididymis, and kidneys of euthanized rats was weighed (after washing away bloodstains in physiological saline, blotting with filter paper, and then weighing), and stored at -80°C for later use.

[0121] Experimental results

[0122] Results of weight and body fat analysis:

[0123] The results of body weight and body fat ratio of mice in the model group, Examples 1-5 and Comparative Examples 1-8 before and after gavage testing are shown in Table 1. The initial body weight is the body weight of the rats after successful high-fat modeling, and the final body weight is the body weight of the rats after 6 weeks of feeding high-fat diet and testing samples.

[0124] Body fat percentage: During sampling, peritesticular and perirenal fat are removed, rinsed with physiological saline, blotted dry with filter paper, and weighed. Body fat percentage = (peritoneal fat + perirenal fat) / final body weight.

[0125] For comparisons of weight gain in Examples 1-5 with that in the model group, ** indicates P < 0.01, and * indicates P < 0.05; for comparisons of body fat percentage in Examples 1-5 with that in the model group, ## indicates P < 0.01, and # indicates P < 0.05; for comparisons of weight gain in Comparative Examples 1-8 with that in Example 1, AA indicates P < 0.01, and A indicates P < 0.05; for comparisons of body fat percentage in Comparative Examples 1-8 with that in Example 1, BB indicates P < 0.01, and B indicates P < 0.05.

[0126] Table 1

[0127]

[0128] Data Analysis:

[0129] Homogeneity of variance analysis was performed on the body weight of 140 mice that successfully underwent high-fat body fat modeling. The box plot of body weight distribution in rats is shown below. Figure 1 The normal PP plot of rat body weight is shown in the figure. Figure 2 The results showed that F=1.165, p=0.313>0.05, indicating no significant difference in initial body weight among the groups. The body weights of the rats in each group were generally distributed between 400g±30g. The PP plot of the initial body weights showed that the data mostly fell along the diagonal of the plot, indicating that the initial body weights of the rats in each group followed a basically normal distribution, and the data distribution was reasonable.

[0130] Based on this, the data from the subsequent high-fat control group (model group), high-fat + Example 1 (Example 1 group), high-fat + Example 2 (Example 2 group), high-fat + Example 3 (Example 3 group), high-fat + Example 4 (Example 4 group), high-fat + Example 5 (Example 5 group), high-fat + Comparative Example 1 (Comparative Example 1 group), high-fat + Comparative Example 2 (Comparative Example 2 group), high-fat + Comparative Example 3 (Comparative Example 3 group), high-fat + Comparative Example 4 (Comparative Example 4 group), high-fat + Comparative Example 5 (Comparative Example 5 group), high-fat + Comparative Example 6 (Comparative Example 6 group), high-fat + Comparative Example 7 (Comparative Example 7 group), and high-fat + Comparative Example 8 (Comparative Example 8 group) have practical reference value.

[0131] Compared with the same model group, the weight gain in Examples 1-5 was highly significant (P < 0.01) and the body fat percentage was significantly different (P < 0.05), indicating that the fermented product obtained by this invention has a good effect on weight loss and fat reduction.

[0132] Compared with Example 1, Comparative Examples 1-8 showed highly significant differences in weight gain (P < 0.01) and significant differences in body fat percentage (P < 0.05 or P < 0.01), indicating that the combined effects of the raw materials and strains in this invention achieved the effect of fat reduction and weight loss.

[0133] Results of inflammatory factor detection:

[0134] The levels of TNF-α, IL-6, and IL-1β inflammatory factors in the blood were detected by enzyme-linked immunosorbent assay (ELISA). The TNF-α ELISA kit (88-7324), IL-1β ELISA kit (88-7064), and IL-6 ELISA kit (88-7013) were all purchased from Ingenium Life Sciences, Inc., USA. The results are shown in Table 2.

[0135] For comparisons of TNF-α in Examples 1-5 with TNF-α in the model group, ** indicates P < 0.01, and * indicates P < 0.05; for comparisons of IL-6 in Examples 1-5 with IL-6 in the model group, ## indicates P < 0.01, and # indicates P < 0.05; for comparisons of IL-1β in Examples 1-5 with IL-1β in the model group, && indicates P < 0.01, and & indicates P < 0.05.

[0136] When comparing the TNF-α of Comparative Examples 1-8 with the TNF-α of Example 1, AA indicates P < 0.01, and A indicates P < 0.05; when comparing the IL-6 of Comparative Examples 1-8 with the IL-6 of Example 1, BB indicates P < 0.01, and B indicates P < 0.05; when comparing the IL-1β of Comparative Examples 1-8 with the IL-1β of Example 1, CC indicates P < 0.01, and C indicates P < 0.05.

[0137] Table 2

[0138]

[0139] Data Result Analysis:

[0140] Compared with the model group, Examples 1-5 showed significant differences in TNF-α, IL-6, and IL-1β, indicating that the fermentation product obtained in this invention can alleviate the body's inflammatory response. Comparative Examples 1-8 showed significant differences in TNF-α, IL-6, and IL-1β compared with Example 1, indicating that the combined action of the various raw materials and strains in this invention achieves an anti-inflammatory effect.

[0141] In summary, the fermented product obtained by this invention has excellent effects on fat reduction, weight loss, and anti-inflammation. It can achieve fat reduction from an anti-inflammatory perspective, truly adjusting metabolic function from within the body and restoring normal fat metabolism.

[0142] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. An anti-inflammatory and fat-reducing fermented product, characterized in that, The fermentation raw materials, by weight, consist of the following components: 150-300 parts of purslane, 20-80 parts of perilla leaves, 10-40 parts of sunflower flowers, 1-10 parts of golden camellia, 0.15-3 parts of fermentation starter powder, and 6-30 parts of enzyme preparation. The fermentation powder is composed of Lactobacillus plantarum, Lactobacillus acidophilus and Bifidobacterium adolescentis, with the weight ratio of each strain being 0.05-0.9:0.09-0.5:0.2-0.6; The enzyme preparation consists of cellulase, pectinase and acidic protease, with a weight ratio of 2-10:2-10:2-10 for each enzyme.

2. The anti-inflammatory and fat-reducing fermented product according to claim 1, characterized in that, The fermentation raw materials, by weight, consist of the following components: 150-200 parts purslane, 20-60 parts perilla leaves, 10-30 parts sunflower flowers, 1-5 parts golden camellia, 0.15-2 parts fermentation starter, and 6-20 parts enzyme preparation.

3. The anti-inflammatory and fat-reducing fermented product according to claim 1, characterized in that, The viable counts of *Lactobacillus plantarum* are 100-300 billion CFU / g, *Lactobacillus acidophilus* are 100-500 billion CFU / g, and *Bifidobacterium adolescentis* are 100-300 billion CFU / g.

4. The anti-inflammatory and fat-reducing fermented product according to claim 1, characterized in that, The cellulase has an activity of 10,000-30,000 U / g, the pectinase has an activity of 10,000-50,000 U / g, and the acidic protease has an activity of 50,000-100,000 U / g.

5. The anti-inflammatory and fat-reducing fermented product according to claim 1, characterized in that, By weight, it consists of the following components: 200 parts purslane, 50 parts perilla leaves, 20 parts sunflower flowers, 5 parts golden camellia, 0.3 parts Lactobacillus plantarum, 0.3 parts Lactobacillus acidophilus, 0.4 parts Bifidobacterium adolescentis, 5 parts cellulase, 5 parts pectinase, and 5 parts acidic protease.

6. The method for preparing the anti-inflammatory and fat-reducing fermented product according to any one of claims 1-5, characterized in that, Includes the following steps: (1) Grind purslane, perilla leaves, sunflower flowers and golden camellia into powder and sieve to a mesh size of 100-600 mesh. Add water and mix evenly, then adjust the pH to 4.0-5.0 to obtain a mixed solution. (2) Add an enzyme preparation to the mixture for enzymatic hydrolysis. After the enzymatic hydrolysis is completed, adjust the pH to 5.0-6.0 to obtain the enzymatic hydrolysate. (3) Extract the enzymatic hydrolysate at 80-102℃ for 10-90 min to obtain the extract; (4) Add fermentation bacteria powder to the extract to ferment and obtain fermentation liquid. Separate the fermentation liquid from solid to liquid to obtain fermentation liquid, which is the fermentation product.

7. The method for preparing the anti-inflammatory and fat-reducing fermented product according to claim 6, characterized in that, The enzymatic hydrolysis conditions in step (2) are a temperature of 40-70℃ and a time of 90-240 min; the fermentation conditions in step (4) are a temperature of 15-45℃ and a time of 36-96 h.

8. The method for preparing the anti-inflammatory and fat-reducing fermented product according to claim 6, characterized in that, In step (1), anhydrous citric acid is used to adjust the pH; in step (2), sodium bicarbonate is used to adjust the pH.

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