A lotus root residue insoluble dietary fiber composition for lowering blood lipid and regulating intestinal flora structure, and a preparation method and application thereof

Abstract

The present application relates to the technical field of health-care medicine, and discloses a lotus root residue insoluble dietary fiber composition for lowering blood lipid and regulating intestinal flora structure, and a preparation method and application thereof. Insoluble dietary fiber in lotus root residue is separated by an enzymatic method, and then mixed with puerarin and xylo-oligosaccharide at a proper ratio to form the lotus root residue insoluble dietary fiber composition. The lotus root residue insoluble dietary fiber composition and lotus root residue insoluble dietary fiber are respectively combined with high-fat feed to feed rats. It is found that the lotus root residue insoluble dietary fiber composition and lotus root residue insoluble dietary fiber can both relieve the obesity symptoms of rats, regulate the intestinal flora structure of rats, maintain the intestinal health of rats, reduce the pathological changes of fat in rats, and have a significant effect of lowering blood lipid and regulating intestinal flora structure. The lotus root residue insoluble dietary fiber composition can be applied to the preparation of health-care products or medicines for regulating the intestinal flora structure of hyperlipidemia.

Description

Technical Field

[0001] This invention relates to the field of health care products technology, specifically to a lotus root residue insoluble dietary fiber composition that lowers blood lipids and regulates intestinal flora structure, as well as its preparation method and application. Background Technology

[0002] The statements in this section provide only background information relevant to the disclosure of this application and may not constitute prior art.

[0003] High blood lipids and fatty liver are caused by long-term intake of high-energy and high-fat diets, coupled with sedentary lifestyles and lack of physical activity. With the continuous improvement of people's living standards, the incidence rate is increasing year by year. Drug treatment often has many adverse reactions and side effects. Health supplements or dietary therapy use gentler methods to gradually alleviate disease symptoms, making them a healthier and more acceptable approach.

[0004] The balance of gut microbiota is closely related to hyperlipidemia and fatty liver. Normal gut microbiota is responsible for converting primary bile acids into secondary bile acids, expressing lipoprotein lipase inhibitors, and producing short-chain fatty acids. A high-fat diet and a high-lipidemia environment directly alter the gut microbiota structure, for example, reducing the number of *Prevotella* and increasing the ratio of Firmicutes to Bacteroides. When the microbiota structure is imbalanced, bile acid metabolism becomes disordered, thus affecting cholesterol synthesis and excretion. Simultaneously, the expression of lipoprotein lipase inhibitors decreases, weakening their inhibitory effect and leading to increased lipoprotein lipase activity. This, in turn, promotes triglyceride deposition in adipose tissue, increases the efficiency of energy absorption in the intestine, exacerbates obesity and metabolic burden, and causes elevated blood lipids and fatty liver. Increasing the dietary fiber content of food helps promote the growth of beneficial gut bacteria and maintain the gut microbiota structure. Dietary fiber refers to the general term for polysaccharides and lignin that are not digested by the human body. It can be divided into two main categories: water-soluble dietary fiber and water-insoluble dietary fiber. Water-soluble dietary fiber mainly consists of storage substances and secretions within plant cells, as well as some microbial polysaccharides and synthetic polysaccharides. Its main components are some gum substances and sugars. The main components of insoluble dietary fiber are cellulose, hemicellulose, lignin, protopectin, and chitosan.

[0005] The prior art disclosed in CN107348351A states that lotus root powder contains abundant mucoprotein and dietary fiber, which can reduce the absorption of fat in the intestines and promote its elimination, thus preventing symptoms of hyperlipidemia; that is, this prior art discloses that lotus root powder can play a role in preventing hyperlipidemia. However, its focus is on the finished lotus root powder. In the process of preparing lotus root powder, after grinding, washing, and filtering, lotus root residue waste and starch-rich lotus root juice are obtained. This part of the lotus root residue waste will be left idle or wasted. With the expansion of lotus root powder production scale, the amount of lotus root residue waste generated is also huge. The reasonable recycling and utilization of this part of lotus root residue waste will greatly increase the added value and economic benefits of lotus root, while also reducing waste and environmental pollution. Summary of the Invention

[0006] The purpose of this invention is to address the problems of large quantities of lotus root residue waste, serious waste, and environmental pollution. It provides a lotus root residue insoluble dietary fiber composition for lowering blood lipids and regulating intestinal flora structure, along with its preparation method and application. This invention utilizes waste resources to prepare a health product with good blood lipid-lowering and intestinal flora-regulating effects, making full use of lotus root resources. It also provides a new type of health product for lowering blood lipids and regulating intestinal flora structure, offering new ideas and methods for preventing, treating, or improving hyperlipidemia and fatty liver.

[0007] The technical solution of the present invention is as follows: This invention provides a lotus root residue insoluble dietary fiber composition that lowers blood lipids and regulates intestinal flora structure, comprising lotus root residue insoluble dietary fiber.

[0008] According to a preferred embodiment, it also includes puerarin and xylooligosaccharides.

[0009] According to a preferred embodiment, the mass ratio of insoluble dietary fiber, puerarin, and xylooligosaccharides in the lotus root residue is 5:5:2.

[0010] According to a preferred embodiment, the insoluble dietary fiber from lotus root residue is prepared by the following method: Step (1): Selection of raw materials: Use lotus root residue with normal color, no odor, and no mold as raw materials; Step (2): Raw material pretreatment: Lotus root residue is mixed evenly with purified water to obtain lotus root residue suspension; Step (3): Pass the lotus root residue suspension through a colloid mill; Step (4): Add cellulase and place in a constant temperature water bath for enzymatic hydrolysis; Step (5): Inactivate the enzymes in the enzymatically hydrolyzed lotus root residue suspension at high temperature. Step (6): Dry the enzyme-inactivated lotus root residue suspension to constant weight; Step (7): Pulverize the dried solid to obtain lotus root residue insoluble dietary fiber powder.

[0011] According to a preferred embodiment, the ratio of lotus root residue to purified water in step (2) is: lotus root residue: purified water = 1:4.

[0012] According to a preferred embodiment, in step (3), the grinding range of the colloid mill is 17-20.

[0013] According to a preferred embodiment, in step (4), the concentration of cellulase added is 0.5% (volume concentration) of the lotus root residue suspension.

[0014] According to a preferred embodiment, in step (5), the high-temperature enzyme inactivation specifically involves placing the enzymatically hydrolyzed lotus root residue suspension in a 100°C drying oven for high-temperature enzyme inactivation for 15 minutes.

[0015] Another aspect of the present invention provides a method for preparing a lotus root residue insoluble dietary fiber composition for lowering blood lipids and regulating intestinal flora structure as described above, comprising the following steps: Step (1): Selection of raw materials: Use lotus root residue with normal color, no odor, and no mold as raw materials; Step (2): Raw material pretreatment: Lotus root residue is mixed evenly with purified water to obtain lotus root residue suspension; Step (3): Pass the lotus root residue suspension through a colloid mill; Step (4): Add cellulase and place in a constant temperature water bath for enzymatic hydrolysis; Step (5): Inactivate the enzymes in the enzymatically hydrolyzed lotus root residue suspension at high temperature. Step (6): Dry the enzyme-inactivated lotus root residue suspension to constant weight; Step (7): Pulverize the dried solid to obtain lotus root residue insoluble dietary fiber powder.

[0016] Another aspect of the present invention provides the application of a lotus root residue insoluble dietary fiber composition for lowering blood lipids and regulating intestinal flora structure as described above in the preparation of health products and pharmaceuticals for regulating the intestinal flora structure of hyperlipidemia.

[0017] Compared with existing technologies, the advantages of this invention are: 1. A lotus root residue insoluble dietary fiber composition for lowering blood lipids and regulating intestinal flora structure. Experiments have shown that the insoluble dietary fiber composition prepared from lotus root residue in this application can significantly reduce the levels of TC, TG, and LDL-C in the serum of HLP rats, and increase the level of HDL-C; reduce liver pathological changes, decrease the levels of TNF-α and IL-6 in serum and liver, increase the level of IL-10, and alleviate inflammation in HLP rats; increase the level of sIgA in the colon and decrease the level of LPS in the colon and serum. 2. A composition of insoluble dietary fiber from lotus root residue that lowers blood lipids and regulates intestinal flora structure, regulating the Firmicutes / Bacteroidetes ratio of intestinal flora, thus modulating the intestinal microecology and reducing the degree of increase in Lactobacillus abundance caused by a high-fat diet. P <0.05) and the degree of reduction in abundance of Lactobacillus spp. and Lactobacillus reuteri ( P <0.05). Attached Figure Description

[0018] Figure 1 The effects of lotus root residue IDF on body weight (A), food intake (B), liver size (C), liver-to-body ratio (D), TC, TG, LLDLD-C, and HDL-C in hyperlipidemic rats (EH). Figure 2 The results of HE staining of (A) colon, (B) liver and (C) liver oil red O staining of hyperlipidemic rats in each group are observed; NC is the normal group, HFD is the high-fat diet group, IDF is the lotus root insoluble dietary fiber group, IDF-PN is the lotus root insoluble dietary fiber and puerarin compound group, PN is the puerarin group and SIM is the simvastatin group. Figure 3 The effect of lotus root residue IDF on inflammation levels in hyperlipidemic rats: (A) ELISA detection of LPS and sIgA levels in the intestines of rats in each group; (B) Q-PCR detection of mRNA expression levels of liver inflammatory factors TNF-α, IL-6, and IL-10 in rats in each group; (C) ELISA detection of serum levels of LPS, IL-6, TNF-α, and IL-10 in rats in each group. Figure 4 The study investigated the effect of lotus root residue IDF on the diversity of gut microbiota in hyperlipidemic rats. Among them, (A), (B), and (C) represent Alpha diversity analysis: Ace, Chao, and Shannon indices, respectively; (D), (E), (F), and (G) represent Beta diversity analysis: PCA, PCoA, NMDS, and sample microbiota typing analysis, respectively. Figure 5The effect of lotus root residue IDF on the taxonomic composition of gut microbiota in hyperlipidemic rats; (A) and (B) are the dysbiosis index and intergroup difference analysis; (C) and (D) are Venn diagrams of species at the genus and species levels; (E), (F), and (G) are Circos diagrams of communities at the phylum, genus, and species levels, respectively. Figure 6 The effect of lotus root residue IDF on the composition of characteristic bacterial strains in the intestine of hyperlipidemic mice; (A) phylum-level community Bar diagram; (B) genus-level community Bar diagram; (C) species-level community Bar diagram; (D) linear discriminant analysis (LDA) bar chart; Figure 7 (A) is a column chart comparing multiple groups at the genus level; (B) is a column chart comparing multiple groups at the species level; (C) is a statistical table of differences between groups of Romboutsia ilealis; (D) is a heatmap of the community at the genus level. Detailed Implementation

[0019] The specific embodiments listed in this invention are merely examples, and the invention is not limited to the specific embodiments described below. For those skilled in the art, any equivalent modifications and substitutions to the embodiments described below are also within the scope of this invention. Therefore, all equivalent transformations and modifications made without departing from the spirit and scope of this invention should be covered within its scope. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. All reagents or instruments whose manufacturers are not specified are commercially available conventional products. To better illustrate this invention, numerous specific details are provided in the following detailed embodiments. Those skilled in the art should understand that this invention can be practiced even without certain specific details. In other embodiments, methods, means, equipment, and steps well known to those skilled in the art are not described in detail in order to highlight the main points of this invention.

[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. Unless otherwise specified, all units used in this specification are International Standard Units (SI), and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors unavoidable in industrial production.

[0021] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0022] I. Experimental Materials: 1. Animals and reagents: 60 SPF-grade male Sprague Dawley (SD) rats, 6 - 8 weeks old, weighing 180 ± 20 g, production license number: SCXK (Beijing) 2024 - 0001, Spiber (Beijing) Technology Co., Ltd.; animal control, high-fat feed, Jiangsu Medison Biomedical Co., Ltd. In this experiment, the animal experiment part follows the principles of the Animal Ethics Committee of Yunnan University of Traditional Chinese Medicine (ethics number R - 062023121). Lotus root residue was purchased from Yunnan Chengjiang Lotus Root Powder Factory in China. Cellulase (100 U / mg) was purchased from Solarbio Co., Ltd., Beijing, China. Cholesterol, 3,5 - dinitrosalicylic acid, salicylic acid, and DPPH were purchased from Sangon Biotech Co., Ltd., Shanghai, China. Kits for total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL - C), and low density lipoprotein cholesterol (LDL - C) were purchased from Nanjing Jiancheng Bioengineering Institute.

[0023] 2. Instruments and equipment: Synergy H1 multi-functional full-wave microplate reader, American Berten Instrumen Company; DHP - 9012 electrothermal constant temperature incubator, Haisenxin Company; HH.S11 - 2 water bath, Shanghai Boxun Medical Biotechnology Co., Ltd.; KH23A high-speed centrifuge, Eppendorf Company; digital slide scanning system (Axioscan 7), ZEISS Company; electrothermal constant temperature forced air drying oven, Shanghai Yuejin Medical Instrument Co., Ltd.

[0024] Example 1 Preparation of insoluble dietary fiber composition from lotus root residue Prepared according to the following method steps: Step (1): Selection of raw materials: Use lotus root residue with normal color, no peculiar smell, and no mildew as raw materials; Step (2): Pretreatment of raw materials: Mix lotus root residue and pure water evenly at a mass ratio of 1:4 to obtain a lotus root residue suspension; Step (3): Pass the lotus root residue suspension through a colloid mill 3 times, and the grinding degree range of the colloid mill is 17 - 20; Step (4): Pour the lotus root residue suspension into an enzymatic hydrolysis tank and add cellulase, set the temperature to a constant 30°C, stir well with a glass rod for enzymatic hydrolysis for 50 minutes, and the enzyme concentration is 1.8% (volume concentration) of the lotus root residue suspension; Step (5): Heat - inactivate the enzyme in the enzymatic hydrolysis tank at 100°C for 15 minutes for the enzymatic hydrolysis - treated lotus root residue suspension; Step (6): Dry the enzyme-inactivated lotus root residue suspension to constant weight; use a vacuum dryer, dry at 80℃ for 100g / 2 hours; Step (7): Pulverize the dried solid to obtain lotus root residue insoluble dietary fiber powder; Step (8): Mix lotus root residue insoluble dietary fiber, puerarin, and xylooligosaccharide in a ratio of 5:5:2 to prepare a lotus root insoluble dietary fiber composition.

[0025] Example 2: Preparation of a high-fat feed formulation with added lotus root residue insoluble dietary fiber. The feed formulations (by mass fraction) for the normal diet group (NC), high-fat diet group (HFD), lotus root insoluble dietary fiber group (IDF), lotus root insoluble dietary fiber and puerarin compound group (IDF-PN), puerarin group (PN), and simvastatin group (SIM) are as follows: Normal group (NC): 77% basal feed, 10% lard, 5% white sugar, 5% egg yolk powder, 2.5% cholesterol, and 0.5% sodium deoxycholate; High-fat diet group (HFD): 85% basal feed, 0.2% propylthiouracil, 7.5% lard, 5% protein powder, 2% cholesterol, and 0.3% sodium deoxycholate; Lotus root insoluble dietary fiber group (IDF): 79.6% basic feed, 0.2% propylthiouracil, 7.5% lard, 5% protein powder, 2% cholesterol, 0.3% sodium deoxycholate, and 5.4% lotus root insoluble dietary fiber powder prepared in step (7) of Example 1.

[0026] Lotus root insoluble dietary fiber puerarin complex (IDF-PN): 79.6% basic feed, 0.2% propylthiouracil, 7.5% lard, 5% protein powder, 2% cholesterol, 0.3% sodium deoxycholate, and 5.4% of the lotus root insoluble dietary fiber composition prepared in step (8) of Example 1.

[0027] Puerarin group (PN): 79.6% basal feed, 0.2% propylthiouracil, 7.5% lard, 5% protein powder, 2% cholesterol, 0.3% sodium deoxycholate, and 5.4% puerarin.

[0028] Xylooligosaccharide group: 79.6% basal feed, 0.2% propylthiouracil, 7.5% lard, 5% protein powder, 2% cholesterol, 0.3% sodium deoxycholate, and 5.4% xylooligosaccharides.

[0029] Simvastatin group (SIM): 77% basal diet, 10% lard, 5% white sugar, 5% egg yolk powder, 2.5% cholesterol, 0.5% sodium cholate, and 2% simvastatin; The following experiments were used to verify the experimental results: 1. Establishment and intervention of hyperlipidemia model The establishment of a high-fat rat model was based on the "Technical Specifications for Testing and Evaluation of Health Foods (2003 Edition)". Sixty male SD rats were acclimatized for one week. Ten rats were randomly selected to continue receiving the basal diet as a blank control group, while the remaining 50 rats were given a high-fat diet as the high-fat model group. The high-fat model group was randomly divided into five subgroups: lotus root insoluble fiber group (IDF), lotus root insoluble fiber combined with kudzu root group (IDF-PN), kudzu root group (PN), simvastatin group (SIM), and model control group (HFD), with 10 rats in each subgroup. Each subgroup was fed the corresponding diet for 7 weeks. Animals were housed in cages of 3-4 per cage, with free access to water and food. Food intake was recorded daily, and body weight was measured every three days. After 7 weeks, serum TC, TG, LDL-C, and HDL-C levels were measured. A significant increase in serum TC or TG in the high-fat rats compared to the blank control group (P<0.05) indicated a successful model.

[0030] After the experimental intervention, rats were fasted for 12 hours, then weighed, anesthetized, and had blood drawn from the abdominal aorta. Blood samples were centrifuged at 3000 r / min for 10 min at 4℃, and the serum was separated and stored at -20℃ for later analysis of various indicators. The liver and colon of the dissected rats were completely removed, and the intestinal contents were used for high-throughput 16S rRNA gene sequencing to detect the intestinal flora. The liver was washed three times with physiological saline, blotted dry with filter paper, photographed, and weighed. Part of the rat liver and colon were routinely fixed in 10% formalin for the preparation of pathological sections, and the remaining tissue was immediately frozen in liquid nitrogen and stored at -80℃ for later analysis of various indicators.

[0031] 2. General activity and liver-to-body ratio in hyperlipidemic rats After the experimental animals were weighed, anesthetized, and dissected, the livers were removed, dried with filter paper, and weighed. The organ-to-body ratio was calculated according to formula (1).

[0032] Visceral ratio (%) = m1 / m2 × 100 Equation (1) In the formula: m1 represents the weight of the rat liver, g; m2 represents the weight of the rat, g.

[0033] The results are as follows Figure 1As shown in the AD diagram, the mice maintained good growth and activity throughout the experimental period, with no injuries or deaths. Compared to the NC group, the HFD group showed a significant increase in both food intake and body weight (P<0.05), indicating that the increase in body weight was related to the high-fat diet. Compared to the HFD group, the IDF and IDF-PN groups had significantly lower body weights, with the IDF-PN group showing even better results (P<0.01), indicating that the intervention of lotus root insoluble dietary fiber effectively inhibited the increase in body weight in hyperlipidemic rats. Compared to the NC group, the HFD group had a significantly higher liver-to-body weight ratio (P<0.05), indicating increased liver weight and hepatomegaly in rats following a high-fat diet. Compared to the HFD group, the IDF and IDF-PN groups had significantly lower liver-to-body weight ratios (P<0.05), indicating that the insoluble dietary fiber from lotus root effectively reduced lipid accumulation and alleviated hepatomegaly.

[0034] 3. Determination of blood lipid levels in hyperlipidemic rats Serum total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured using the corresponding commercial kits (Nanjing Jiancheng Biotechnology Institute, Nanjing, China) according to the manufacturer's instructions.

[0035] Result: As Figure 1 As shown in EF, compared with the NC group, the serum levels of TC, TG, LDL-C, and HDL-C in the HFD group were significantly increased (P<0.05); compared with the HFD group, the serum levels of TC, TG, LDL-C, and HDL-C in the IDF group and IDF-PN group were significantly lower than those in the HFD group, with the IDF-PN group showing better results (P<0.05), indicating that the intervention of lotus root insoluble dietary fiber can effectively reduce the blood lipid levels of hyperlipidemic rats.

[0036] 4. Pathological observation of colon and liver tissue sections of hyperlipidemic rats First, paraffin sections were dewaxed sequentially in xylene, anhydrous ethanol, and alcohol until water was obtained. Then, hematoxylin staining was performed. After hematoxylin staining, the sections were dehydrated and then stained with eosin for 5 minutes. Finally, they were dehydrated sequentially in anhydrous ethanol, n-butanol, and xylene. After drying and mounting, images were acquired and analyzed under a microscope. Frozen sections were thawed, fixed, and dried again. Then, they were immersed in oil red stain for 8-10 minutes (away from light), differentiated with 75% alcohol, rinsed with deionized water, stained with hematoxylin for 3-5 minutes, blued with ammonia, rinsed with deionized water, mounted with glycerol gelatin, and the results were photographed and analyzed.

[0037] The results are as follows Figure 2As shown: HE-stained pathological sections of rat livers in each group: Compared with the NC group, the HFD group showed vacuolar changes and inflammatory cell infiltration in the liver tissue, indicating that the model was successfully established; Compared with the HFD group, the IDF group and IDF-PN group showed a reduction in vacuolar lesions in the liver tissue, indicating that the intervention of lotus root insoluble dietary fiber can significantly reduce vacuolar changes and inflammatory cell infiltration in the liver tissue.

[0038] Oil Red O staining pathological sections of rat livers in each group: Compared with the NC group, the HFD group showed a significant increase in liver lipid droplet area (P<0.05), indicating successful model establishment; Compared with the HFD group, the IDF group and IDF-PN group showed significantly lower liver lipid droplet area, with the IDF-PN group showing better results (P<0.05), indicating that lotus root insoluble dietary fiber can significantly reduce lipid droplet deposition in liver tissue after intervention; HE staining pathological sections of rat colons in each group: In the NC group, the intestinal mucosa and mucus layer were intact, the number of goblet cells was normal, the intestinal mucosa was undamaged, and there was no obvious inflammatory cell infiltration; Compared with the NC group, the HFD group showed intestinal mucosal damage, fewer goblet cells, more intestinal mucosal defects, and large areas of inflammatory infiltration; Compared with the HFD group, the IDF group and IDF-PN group showed increased goblet cell number, better recovery of intestinal mucosal defects, and reduced inflammatory cell infiltration.

[0039] 5. Effects of lotus root residue IDF on LPS and sIgA levels in the intestines of hyperlipidemic rats, expression of hepatic inflammatory factors TNF-α, IL-6, and IL-10 mRNA, and serum levels of LPS, IL-6, TNF-α, and IL-10. The concentrations of TNF-α, IL-10, IL-6, and LPS in mouse serum were detected using an ELISA kit (Enzyme-Label Biotechnology Co., Ltd., Jiangsu, China). Strips were removed from the aluminum foil bag after equilibration at room temperature for 20 min. Standard wells and sample wells were set up. 50 μl of different concentrations of standard solution were added to each standard well. 10 μl of the test sample was added to each sample well, followed by 40 μl of sample diluent. Except for the blank wells, 100 μl of horseradish peroxidase (HRP)-labeled detection antibody was added to each standard and sample well. The plates were sealed and incubated at 37°C for 1 h. The liquid in the wells was discarded, and the plates were patted dry on absorbent paper. Washing buffer was added to each well, and the plates were allowed to stand for 1 minute. The wash buffer was then shaken off, and the plates were patted dry on absorbent paper. This process was repeated 5 times. 50 μl each of substrate A and B were added to each well, and the plates were incubated in the dark at 37°C for 15 min. Stop solution was added, and the OD value of each well was measured at 450 nm within 15 min.

[0040] The results are as follows Figure 3As shown: ELISA detection of LPS and sIgA levels in the intestines of rats in each group: Compared with the NC group, the LPS content in the intestines of rats in the HFD group was significantly increased (P<0.05), while the sIgA content was significantly decreased (P<0.05); Compared with the HFD group, the LPS content in the intestines of rats in the IDF group and IDF-PN group was significantly decreased (P<0.05), while the sIgA content was significantly increased (P<0.05), indicating that the intervention of lotus root insoluble dietary fiber can effectively reduce the level of intestinal inflammation in HLP rats and repair intestinal mucosal barrier damage. Q-PCR was used to detect the expression levels of hepatic inflammatory factors TNF-α, IL-6, and IL-10 mRNA in rats of different groups. Compared with the NC group, the expression levels of hepatic inflammatory factors TNF-α and IL-6 mRNA in the HFD group were significantly increased (P<0.05), while the expression level of IL-10 mRNA was significantly decreased (P<0.05). Compared with the HFD group, the expression levels of hepatic inflammatory factors TNF-α and IL-6 mRNA in the IDF group and IDF-PN group were significantly decreased (P<0.05), while the expression level of IL-10 mRNA was significantly increased (P<0.05). This indicates that the intervention of lotus root insoluble dietary fiber can effectively reduce the level of hepatic inflammation in HLP rats.

[0041] ELISA was used to detect the levels of LPS, IL-6, TNF-α, and IL-10 in the serum of rats in each group. Compared with the NC group, the levels of LPS, IL-6, and TNF-α in the serum of the HFD group increased (P<0.05), while the level of IL-10 decreased (P<0.05). Compared with the HFD group, the levels of LPS, IL-6, and TNF-α in the serum of the IDF group and the IDF-PN group decreased (P<0.05), while the level of IL-10 increased (P<0.05), indicating that the intervention of lotus root insoluble dietary fiber can regulate inflammation in HLP rats.

[0042] 6. Data Statistical Analysis All experimental data are expressed as mean ± standard deviation (x ± s) and statistical analysis was performed using GraphPad software.

[0043] Analysis of variance was used for comparisons between groups, and a p-value < 0.05 was considered statistically significant.

[0044] II. Lotus root insoluble dietary fiber composition regulates the structure and abundance of intestinal flora. Alpha diversity analysis reflects the richness and diversity of the microbial community; ACE and Chao1 indices reflect changes in the relative abundance of the microbial community; the shanoon index reflects changes in microbial community diversity; Beta analysis reflects the magnitude of differences between microbial groups; Principal Component Analysis (PCA) and Non-metric Multidimensional Scaling (NMDS) represent the magnitude of differences between sample groups based on the distance between them—closer distances indicate greater similarity, and greater distances indicate greater differences; the Muscle Dysbiosis Index (MDI) is an index that determines the degree of microbial ecological dysbiosis. A higher value indicates a greater degree of microbial dysbiosis.

[0045] The results are as follows Figure 4 and Figure 5 As shown, IDF and IDF-PN can increase the abundance of gut microbiota and reduce the diversity of gut microbiota. After IDF and IDF-PN intervention, the composition of gut microbiota tends to be similar to that of the control group, with little difference in composition, and the dysbiosis index and inter-group differences are reduced. At the genus level, there are 20 common genera among the groups, and at the species level, there are 24 common species among the groups. The composition of dominant microbiota in gut contents changes at the phylum, genus and species levels.

[0046] like Figure 6 and Figure 7 As shown, compared with the blank group, the abundance of Lactobacillus and other bacteria in the model group was significantly increased. P <0.05), the abundance of bacteria such as *Lactobacillus reuteri* and *Lactobacillus reuteri* was significantly reduced ( P <0.05); compared with the model group, the abundance of Lactobacillus species was significantly reduced after IDF and IDF-PN sample intervention ( P <0.05), the abundance of *Lactobacillus reuteri* and *Lactobacillus reuteri* significantly increased ( P <0.05).

[0047] Changes in genus-level bacterial community structure: NC Group: Lactobacillus 28%, Romboutsia 27% Limosilactobacillus 19%; HFD group : Lactobacillus 44%, Limosilactobacillus 12% Ligilactobacillus 9%; PN group: Lactobacillus 30% Romboutsia 25%, Limosilactobacillus 16%; IDF Group: Limosilactobacillus 20% Lactobacillus 19%Romboutsia 15%; IDF-PN group: Lactobacillus 34%, Romboutsia twenty one%, Limosilactobacillus 16%; SIM group: Romboutsia 35%, Lactobacillus 28%, Limosilactobacillus 11%.

[0048] Changes in microbial community structure at the species level: NC Group: Romboutsia ilealis 27% Limosilactobacillus reuteri 14% Lactobacillus johnsonii 13%; HFD group: Lactobacillus_acidophilus twenty four%, Lactobacillus hominis 8% Lactobacillus johnsonii 8%; PN group: Romboutsia ilealis 25%, Limosilactobacillus reuteri 12% Lactobacillus intestinalis 11%; IDF Group: Romboutsia ilealis 15% Limosilactobacillus reuteri 16% Lactobacillus intestinalis 9%; IDF-PN group: Romboutsia ilealis twenty one%, Lactobacillus intestinalis 15% Lactobacillus johnsonii 14%; SIM group: Romboutsia ilealis 34%, Lactobacillus johnsonii 11% Lactobacillus johnsonii 11%.

[0049] Differential bacterial genera were screened as Romboutsia The strain is Romboutsia ilealis.

[0050] The embodiments described above merely illustrate specific implementation methods of this application, and while the descriptions are detailed and specific, they should not be construed as limiting the scope of protection of this application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the technical solution of this application, and these modifications and improvements all fall within the scope of protection of this application.

Claims

1. A lotus root residue insoluble dietary fiber composition for lowering blood lipids and regulating intestinal flora structure, characterized in that, This includes insoluble dietary fiber from lotus root residue.

2. A lotus root residue insoluble dietary fiber composition according to claim 1 for lowering blood lipid and regulating intestinal flora structure, characterized in that, It also includes puerarin and xylooligosaccharides.

3. A lotus root residue insoluble dietary fiber composition according to claim 2 for lowering blood lipid and regulating intestinal flora structure, characterized in that, The mass ratio of insoluble dietary fiber, puerarin, and xylooligosaccharides in the lotus root residue is 5:5:

2.

4. The lotus root residue insoluble dietary fiber composition according to claim 1, characterized in that, The insoluble dietary fiber from lotus root residue was prepared according to the following method: Step (1): Selection of raw materials: Use lotus root residue with normal color, no odor, and no mold as raw materials; Step (2): Raw material pretreatment: Lotus root residue is mixed evenly with purified water to obtain lotus root residue suspension; Step (3): Pass the lotus root residue suspension through a colloid mill; Step (4): Add cellulase and place in a constant temperature water bath for enzymatic hydrolysis; Step (5): Inactivate the enzymes in the enzymatically hydrolyzed lotus root residue suspension at high temperature. Step (6): Dry the enzyme-inactivated lotus root residue suspension to constant weight; Step (7): Pulverize the dried solid to obtain lotus root residue insoluble dietary fiber powder.

5. A lotus root residue insoluble dietary fiber composition according to claim 4, characterized in that, In step (2), the ratio of lotus root residue to purified water is 1:

4.

6. A dietary fiber composition of lotus root residue insoluble for lowering blood lipid and regulating intestinal flora structure according to claim 4, characterized in that, In step (3), the grinding range of the colloid mill is 17-20.

7. A lotus root residue insoluble dietary fiber composition according to claim 4, characterized in that, In step (4), the concentration of cellulase added is 0.5% of the lotus root residue suspension.

8. A dietary fiber composition of lotus root residue insoluble for lowering blood lipid and regulating intestinal flora structure according to claim 4, characterized in that, In step (5), the high-temperature enzyme inactivation is specifically performed by placing the enzymatically hydrolyzed lotus root residue suspension in a 100°C drying oven for 15 minutes to inactivate the enzyme.

9. The method for preparing a lotus root residue insoluble dietary fiber composition for lowering blood lipids and regulating intestinal flora structure as described in claim 1, characterized in that, Includes the following steps: Step (1): Selection of raw materials: Use lotus root residue with normal color, no odor, and no mold as raw materials; Step (2): Raw material pretreatment: Lotus root residue is mixed evenly with purified water to obtain lotus root residue suspension; Step (3): Pass the lotus root residue suspension through a colloid mill; Step (4): Add cellulase and place in a constant temperature water bath for enzymatic hydrolysis; Step (5): Inactivate the enzymes in the enzymatically hydrolyzed lotus root residue suspension at high temperature. Step (6): Dry the enzyme-inactivated lotus root residue suspension to constant weight; Step (7): Pulverize the dried solid to obtain lotus root residue insoluble dietary fiber powder.

10. The application of the lotus root residue insoluble dietary fiber composition according to any one of claims 1-8 in the preparation of health products and pharmaceuticals for regulating the intestinal flora structure of hyperlipidemia.

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