Application of corn cob xylooligosaccharides in regulating glucose and lipid metabolism

Abstract

This invention discloses the application of corn cob xylooligosaccharides in regulating glucose and lipid metabolism, belonging to the field of pharmaceutical technology. The corn cob xylooligosaccharides (XO) of this invention can regulate glucose metabolism levels, improve dyslipidemia, inhibit abnormal expression of inflammatory factors, and reduce oxidative stress damage in the body. It is a natural intervention strategy that achieves multi-pathway and multi-target synergistic regulation of glucose and lipid metabolism disorders, comprehensively improving the pathological state caused by these disorders. This overcomes the problem that existing treatment strategies for glucose and lipid metabolism disorders often focus on single targets and are unable to simultaneously intervene in multiple pathological processes such as glucose metabolism disorders, dyslipidemia, inflammatory responses, and oxidative stress. This provides reliable experimental evidence and theoretical support for the further development (development as a natural active ingredient for adjuvant treatment of glucose and lipid metabolism disorders) and application of corn cob xylooligosaccharides in glucose and lipid metabolism disorders.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the application of corn cob xylooligosaccharides in regulating glucose and lipid metabolism. Background Technology

[0002] Glucose and lipid metabolism disorders are a group of metabolic syndromes characterized by hyperglycemia, dyslipidemia, insulin resistance, and obesity. They are important pathological bases for type 2 diabetes, non-alcoholic fatty liver disease, atherosclerosis, and cardiovascular and cerebrovascular diseases. With changes in dietary structure and lifestyle, the incidence of diseases related to glucose and lipid metabolism disorders continues to rise globally, becoming a significant public health burden. Currently, clinical interventions for glucose and lipid metabolism disorders mainly rely on chemical drugs, such as biguanides (metformin), thiazolidinediones, statins, and fibrates. However, long-term use often has limitations such as gastrointestinal adverse reactions, liver and kidney damage, hypoglycemia risk, and drug tolerance. In addition, functional polysaccharides based on natural products are becoming a research hotspot in the field of metabolic disease intervention due to their advantages of multi-target, low toxicity, and comprehensive regulation. Among them, β-glucan and arabinoxylan from cereals have been proven to have significant effects on improving blood glucose and blood lipids.

[0003] The current research focus in the field of metabolic disease prevention and control is to discover novel substances that are readily available, safe and reliable, and can act on multiple targets, thereby regulating glucose and lipid metabolism, fundamentally correcting metabolic disorders, and delaying the development of chronic diseases. Summary of the Invention

[0004] The purpose of this invention is to provide the application of corn cob xylooligosaccharides in regulating glycolipid metabolism, so as to solve the problems existing in the prior art.

[0005] To achieve the above objectives, the present invention provides the following solution: One of the technical solutions of this invention is the application of corn cob xylooligosaccharides in the preparation of drugs that regulate glucose and lipid metabolism disorders.

[0006] Preferably, the symptoms of the glucose and lipid metabolism disorder include one or more of the following: glucose metabolism disorder, lipid metabolism disorder, inflammatory response, and oxidative stress damage.

[0007] Preferably, the concentration of the corn cob xylooligosaccharide used is 125~500 μg / mL.

[0008] Corn cob xylooligosaccharides can regulate glucose metabolism, improve the disordered state of glucose metabolism in glucose and lipid metabolism disorder models, and reduce the metabolic burden of glucose toxicity on the body; they can regulate lipid metabolism disorders caused by glucose and lipid metabolism disorders, reduce the levels of blood lipid-related indicators such as triglycerides and free fatty acids, and reduce cell damage; they can effectively inhibit chronic inflammatory responses under glucose and lipid metabolism disorder conditions and reduce the levels of inflammatory factors; they can effectively reduce oxidative stress damage related to glucose and lipid metabolism disorders and enhance the body's antioxidant defense capabilities.

[0009] The corn cob xylooligosaccharide used in this invention is a natural active ingredient from corn cob, an agricultural byproduct. Using it to treat disorders of glucose and lipid metabolism can realize the resource utilization of agricultural waste.

[0010] The second technical solution of the present invention: the application of corn cob xylooligosaccharides in the preparation of drugs that improve glucose metabolism disorders.

[0011] The third technical solution of the present invention: the application of corn cob xylooligosaccharides in the preparation of drugs that reduce the content of triglycerides and / or free fatty acids.

[0012] The fourth technical solution of the present invention: the application of corn cob xylooligosaccharides in the preparation of drugs that inhibit inflammatory responses caused by disorders of glycolipid metabolism.

[0013] The fifth technical solution of the present invention: the application of corn cob xylooligosaccharides in the preparation of drugs that alleviate oxidative stress damage caused by glucose and lipid metabolism disorders.

[0014] The sixth technical solution of the present invention: a pharmaceutical composition for treating disorders of glucose and lipid metabolism, comprising corn cob xylooligosaccharides and a pharmaceutically acceptable carrier.

[0015] Preferably, the dosage form of the pharmaceutical composition includes an oral formulation or an injectable formulation.

[0016] The advantages of this invention are as follows: (1) Pioneering a new application of xylooligosaccharides from corn cobs in regulating glucose and lipid metabolism disorders This invention utilizes corn planting byproducts as raw materials, belonging to the category of natural non-grain bio-based raw materials. It transforms discarded corn cobs after harvest into high-value resources, turning agricultural waste into high-end functional sugars, with no secondary pollution during production. The extracted byproducts can also be used in organic fertilizers, edible mushroom substrates, and biomass fuels, achieving a "zero-waste" cycle and significant carbon emission reduction benefits. This invention overcomes the limitations of traditional nutritional or basic activity studies of corn cob xylooligosaccharides. Starting from the cellular level, it is the first to systematically demonstrate the significant role of corn cob xylooligosaccharides in improving glucose and lipid metabolism disorders. It provides a new natural polysaccharide candidate intervention for the prevention and treatment of glucose and lipid metabolism disorders, expanding the application prospects of corn cob xylooligosaccharides in regulating glucose and lipid metabolism disorders.

[0017] (2) Provide experimental evidence for multiple targets This invention constructs a cellular-level evaluation system, demonstrating from the perspectives of metabolic state and cell damage that corn cob xylooligosaccharides can improve glucose metabolism abnormalities, lipid metabolism disorders, chronic inflammatory responses, and oxidative stress related to glucose and lipid metabolism disorders, providing multi-level and reliable experimental evidence for their improvement of the pathological state of glucose and lipid metabolism disorders.

[0018] (3) Clarify the dose-response relationship Through multi-dose gradient design, the effective range of xylooligosaccharides under in vitro conditions (125~500 μg / mL) was clarified, and it was confirmed that xylooligosaccharides have obvious dose-dependent characteristics in improving glucose and lipid metabolism disorders, providing experimental basis for subsequent functional formulation development and application dosage design.

[0019] (4) Achieving the advantages of multi-target synergistic regulation Corn cob xylooligosaccharides can simultaneously regulate multiple key pathological processes caused by glucose and lipid metabolism disorders, such as glucose metabolism, lipid metabolism, inflammatory response, and oxidative stress. Through multi-target and multi-pathway synergistic regulation, they can improve the metabolic microenvironment as a whole, which is more conducive to the comprehensive management of complex metabolic diseases than single-target intervention strategies.

[0020] (5) It combines safety and long-term intervention potential Corn cob xylooligosaccharides, derived from corn cobs, a byproduct of corn cultivation, exhibit good biocompatibility and safety, making them suitable for long-term intervention and management of glucose and lipid metabolism disorders. They function by improving metabolic imbalances and alleviating upstream pathological processes such as chronic inflammation and oxidative stress, demonstrating potential value in improving glucose and lipid metabolism disorders from a pathological perspective.

[0021] (6) The experimental system is standardized and reliable, and has good prospects for transformation. The cell models, evaluation indicators, and detection methods used in this invention are all widely used or recognized reliable methods in the field. The experimental design is scientific and rigorous, and the data system is complete and logically clear, which provides a strong guarantee for the reliability and reproducibility of the research conclusions. It also lays a solid technical foundation for the further development and application of corn cob xylooligosaccharides in the field of regulating glucose and lipid metabolism disorders.

[0022] The present invention discloses the following technical effects: (1) The corn cob xylooligosaccharide (XO) of the present invention can regulate blood glucose metabolism, improve blood lipid abnormalities, inhibit the abnormal expression of inflammatory factors, and reduce oxidative stress damage in the body, thereby alleviating glucose and lipid metabolism disorders. It is a natural intervention solution that realizes multi-pathway and multi-target synergistic regulation of glucose and lipid metabolism disorders, and improves the pathological state of glucose and lipid metabolism disorders as a whole. It overcomes the problem that existing glucose and lipid metabolism disorder treatment strategies are mostly based on single targets and are difficult to intervene in multiple pathological links such as glucose metabolism disorders, blood lipid abnormalities, chronic inflammation and oxidative stress at the same time. It provides reliable experimental basis and theoretical support for the further development (development as a natural active ingredient for the adjuvant treatment of glucose and lipid metabolism disorders) and application of corn cob xylooligosaccharide in the prevention and treatment of glucose and lipid metabolism disorders.

[0023] (2) This invention provides a novel pharmaceutical use for corn cob xylooligosaccharide (XO) derived from corn cob. Using this corn cob xylooligosaccharide (XO) can effectively improve or treat disorders of glucose and lipid metabolism, filling the research gap in the prior art that lacks systematic evaluation of the role of corn cob xylooligosaccharide in improving and regulating abnormal glucose and lipid metabolism, and expanding the application scenarios of natural active ingredients derived from agricultural by-products in the prevention and treatment of disorders of glucose and lipid metabolism. Attached Figure Description

[0024] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0025] Figure 1 This refers to the intracellular glycogen content after the intervention in Example 1. Figure 2 This represents the amount of glucose consumed by cells after the intervention in Example 1. Figure 3 The FFA content in the cell culture supernatant after the intervention in Example 1; Figure 4 The FFA content in the cell homogenate after the intervention in Example 1; Figure 5 The TC content in the cell culture supernatant after the intervention in Example 1; Figure 6 The TC content in the cell homogenate after the intervention in Example 1; Figure 7 The TG content in the cell culture supernatant after the intervention in Example 1; Figure 8 The TG content in the cell homogenate after the intervention in Example 1; Figure 9 This refers to the IL-1β concentration in the cell homogenate after the intervention in Example 1. Figure 10 This refers to the IL-6 concentration in the cell homogenate after the intervention in Example 1. Figure 11 This refers to the TNF-α concentration in the cell homogenate after the intervention in Example 1. Figure 12 The content of GSH-PX in the cell culture supernatant after the intervention in Example 1; Figure 13 The content of GSH-PX in the cell homogenate after the intervention in Example 1; Figure 14 The MDA content in the cell culture supernatant after the intervention in Example 1; Figure 15 The content of MDA in the cell homogenate after the intervention in Example 1; Figure 16 The SOD content in the cell culture supernatant after the intervention in Example 1; Figure 17 The SOD content in the cell homogenate after the intervention in Example 1. Detailed Implementation

[0026] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0027] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0028] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0029] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0030] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0031] It should be noted that any aspects not described in detail in this invention are conventional practices in the field and are not the focus of this invention.

[0032] The corn cob xylooligosaccharides used in the specific embodiments of this invention were purchased from Lanzhou Waterles Biotechnology Co., Ltd.

[0033] Example 1 Application of corn cob xylooligosaccharides in regulating glucose and lipid metabolism disorders: (I) Establishment of cell model and drug administration Experimental cells: HepG2 cells.

[0034] HepG2 cells were cultured in complete medium (containing 5.5 mmol / L glucose, 10% fetal bovine serum and 1% penicillin-streptomycin solution) and routinely cultured in a 37°C, 5% CO2 incubator.

[0035] A cell model related to glucose and lipid metabolism disorders was established using high-glucose and high-fat stimulation. Cell groups were as follows: Normal control group (NC): complete culture medium; Model group (MC): 25 mM glucose (HG) + 200 μM palmitic acid (PA) high glucose and high fat medium; Low-dose corn cob xylooligosaccharide group (XL): 25 mM glucose + 200 μM palmitic acid high-sugar high-fat medium + 125 μg / mL corn cob xylooligosaccharide; Medium-dose group of corn cob xylooligosaccharide (XM): 25 mM glucose + 200 μM palmitic acid high sugar and high fat medium + 250 μg / mL corn cob xylooligosaccharide; High-dose corn cob xylooligosaccharide group (XH): 25 mM glucose + 200 μM palmitic acid high-sugar high-fat medium + 500 μg / mL corn cob xylooligosaccharide.

[0036] (ii) Corn cob xylooligosaccharides improve blood glucose metabolism By detecting the level of glycogen synthesis and glucose consumption capacity in HepG2 cells induced by high glucose and high lipid, the regulatory effect of corn cob xylooligosaccharide (XO) on cellular glucose metabolism disorders was evaluated, clarifying its biological effects of promoting glucose utilization and improving glycogen synthesis, and providing cellular experimental evidence for its intervention in abnormal blood glucose.

[0037] (1) The intervention experiment was divided into three phases: The 0-24 hour period is the logarithmic growth phase, during which cells (1×10⁻⁶) grow at a rate of 10⁻⁶. 4 (cells / mL) were inoculated into complete culture medium; The model induction period was 24-48 hours. The normal control group (NC) was given complete culture medium, while the model group (MC) and the intervention group (low-dose corn cob xylooligosaccharide group (XL), medium-dose corn cob xylooligosaccharide group (XM), and high-dose corn cob xylooligosaccharide group (XH)) were given high-sugar and high-fat culture medium (25 mM glucose + 200 μM palmitic acid). The intervention period was 48–72 hours. The normal control group (NC) was given complete culture medium; the model group (MC) was given high-sugar, high-fat culture medium (25 mM glucose + 200 μM palmitic acid); the low-dose corn cob xylooligosaccharide group (XL) was given high-sugar, high-fat culture medium containing 125 μg / mL corn cob xylooligosaccharide (25 mM glucose + 200 μM palmitic acid + 125 μg / mL corn cob xylooligosaccharide); the medium-dose corn cob xylooligosaccharide group (XM) was given high-sugar, high-fat culture medium containing 250 μg / mL corn cob xylooligosaccharide (25 mM glucose + 200 μM palmitic acid + 250 μg / mL corn cob xylooligosaccharide); and the high-dose corn cob xylooligosaccharide group (XH) was given high-sugar, high-fat culture medium containing 500 μg / mL corn cob xylooligosaccharide (25 mM glucose + 200 μM palmitic acid + 500 μg / mL corn cob xylooligosaccharide). μg / mL corn cob xylooligosaccharides).

[0038] (2) Measurement of intracellular glycogen content Discard the old culture medium from the culture plate, gently wash the cells twice with pre-cooled PBS, and aspirate any remaining liquid. Digest the cells with trypsin to prepare a cell suspension, centrifuge at 1000 rpm for 10 min, discard the supernatant, and retain the cell pellet. Add 0.5–1 mL of PBS to the cell pellet, mix gently, centrifuge at 1000 rpm for 10 min, discard the supernatant, and retain the cell pellet.

[0039] Add 0.2–0.5 mL of PBS to the cell pellet, sonicate or homogenize manually under ice-water bath conditions, without centrifugation, and directly take a sample (take a portion to measure the total protein concentration). Add 0.05 mL of the sample to 0.15 mL of alkaline solution, boil in a water bath for 20 min, and cool with running water to obtain the glycogen detection solution.

[0040] The reaction system was prepared according to the instructions using a glycogen assay kit (anthrone colorimetric method), and the absorbance value at a specific wavelength (620 nm) was measured using an ELISA reader.

[0041] Total cellular protein concentration was simultaneously determined using the BCA method. Glycogen (GY) content was standardized to "mg / mgprot" to eliminate differences in cell number. Results are shown below. Figure 1 .

[0042] (3) Detection of cellular glucose consumption Before the intervention began, freshly prepared culture media were collected from each group as “initial glucose concentration control”; Twenty-four hours after the intervention, the cell culture supernatant of each group was accurately collected and centrifuged at 4°C and 3000 rpm for 5 min to remove cell debris. The glucose (GLU) concentration in the initial culture medium and the supernatant after intervention was determined using a glucose oxidase assay kit. Calculate glucose consumption: Glucose consumption (GC, mmol / L) = Initial glucose concentration - Post-intervention glucose concentration, reflecting the cell's ability to utilize glucose. Results are shown in [link to results]. Figure 2 .

[0043] from Figures 1-2 As can be seen from the results of the blood glucose metabolism-related index detection, compared with the normal control group (NC), the HepG2 cells in the high glucose and high fat model group (MC) induced by high glucose (HG) and palmitic acid (PA) showed a significant decrease in intracellular glycogen (GY) synthesis and a significant reduction in glucose consumption (GC). This indicates that the high glucose and high fat environment successfully caused cellular blood glucose metabolism disorder, impaired glucose utilization, inhibited glycogen synthesis, and intracellular glucose accumulation.

[0044] After intervention with different concentrations of corn cob xylooligosaccharides (125 μg / mL, 250 μg / mL, and 500 μg / mL), the above-mentioned blood glucose metabolism indicators of XL, XM, and XH all showed a significant dose-dependent improvement trend. Among them, compared with the model group (MC), the medium-dose group (XM) and the high-dose group (XH) showed significantly increased intracellular glycogen (GY) content (P<0.01) and significantly increased glucose consumption (GC) in the culture medium (P<0.01), with the high-dose group showing the most significant improvement.

[0045] The above results indicate that xylooligosaccharides from corn cobs can promote glycogen synthesis in HepG2 cells induced by high glucose and high lipids in a dose-dependent manner, enhance the cells' ability to take up and utilize glucose, reduce intracellular glucose accumulation, and effectively improve cellular glucose metabolism disorders, providing reliable cellular experimental evidence for its potential role as a glucose regulation intervention substance.

[0046] (III) Corn cob xylooligosaccharides improve three blood lipid parameters By detecting cholesterol (TC), triglycerides (TG), and free fatty acids (FFA) in HepG2 insulin-resistant cells induced by high glucose and high lipid levels, this study evaluated the effect of corn cob xylooligosaccharide (XO) on improving glucose and lipid metabolism disorders in insulin-resistant cells, providing cellular experimental evidence for its ability to improve abnormal glucose and lipid metabolism by regulating hepatocyte lipid metabolism.

[0047] Using the same intervention experiment as described above, the following glucose metabolism-related indicators were measured after the intervention period ended: (1) After the intervention, cell culture supernatant and cell samples were collected separately, as follows: Cell culture supernatant collection: Carefully aspirate the culture supernatant, centrifuge at 3000 rpm for 15 min at 4℃ to remove cell debris, collect the clear supernatant, aliquot and store at -80℃ for later use in detecting cholesterol (TC), triglycerides (TG) and free fatty acids (FFA).

[0048] Cell sample preparation: Take the prepared cell suspension, centrifuge at 1000 rpm for 10 min, discard the supernatant, and retain the cell pellet; wash twice with PBS (pH 7.4), centrifuge at 1000 rpm for 10 min again, discard the supernatant, and retain the cell pellet; Cell disruption: add 0.2–0.3 mL of PBS (pH 7.4) for homogenization, and sonicate under ice-water bath conditions (300 W power, 3–5 seconds / batch, 30-second interval, repeated 3–5 times). The prepared homogenate is directly measured without centrifugation. Aliquot the cell homogenate and store at -80℃.

[0049] (2) The contents of cholesterol (TC), triglycerides (TG), and free fatty acids (FFA) in cell culture supernatant and cell homogenate were detected using a microplate reader method, strictly following the instructions of the corresponding biochemical assay kits. The absorbance values ​​were read, and the concentrations of each index were calculated based on the standard curve to evaluate the ameliorative effect of corn cob xylooligosaccharides on lipid metabolism disorders induced by high sugar and high lipid levels in HepG2 cells. The results are shown in […]. Figures 3-8 , Figure 3 The content of FFA in cell culture supernatant. Figure 4 This refers to the FFA content in cell homogenate. Figure 5 The TC content in the cell culture supernatant. Figure 6 The TC content in cell homogenate. Figure 7 This refers to the TG content in the cell culture supernatant. Figure 8 This represents the TG content in the cell homogenate.

[0050] from Figures 3-8The biochemical test results showed that, compared with the normal control group (NC), the intracellular free fatty acid (FFA), total cholesterol (TC), and triglyceride (TG) content of HepG2 cells in the high-sugar and high-fat induced model group (MC) was significantly increased (P<0.01). This indicates that the high-sugar and high-fat environment successfully induced significant lipid accumulation in HepG2 cells, and at the same time, the cells showed a damaging response, simulating the pathological state of glucose and lipid metabolism disorder and cell damage.

[0051] After intervention with different concentrations of corn cob xylooligosaccharides (125 μg / mL, 250 μg / mL, and 500 μg / mL), the intracellular FFA, TC, and TG contents in the low-dose group (XL), medium-dose group (XM), and high-dose group (XH) all showed a dose-dependent decreasing trend. Among them, the intracellular FFA and TC contents in the medium-dose group (XM) and high-dose group (XH) were significantly lower than those in the model group (PA) (P<0.05). Although the TG content showed a significant decreasing trend compared with the low-dose group (XL), the differences in some indicators did not reach a statistically significant level.

[0052] The above results indicate that xylooligosaccharides from corn cobs can effectively improve lipid metabolism abnormalities induced by high sugar and high lipid levels in HepG2 cells, while also alleviating cell damage caused by high sugar and high lipid levels. They can reduce the accumulation of intracellular FFA, TC, and TG in a dose-dependent manner, thereby playing a regulatory role in glucose and lipid metabolism disorders and a protective role in cells.

[0053] (iv) Effects of corn cob xylooligosaccharides on inflammatory factors By detecting the levels of inflammatory factors interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in HepG2 cells induced by high glucose and high lipid levels, this study aimed to evaluate the regulatory effect of xylooligosaccharide (XO) from corn cobs on chronic inflammatory responses under conditions of disordered glucose and lipid metabolism, and to provide experimental evidence for its role in improving the inflammatory imbalance caused by disordered glucose and lipid metabolism.

[0054] Using the same intervention experiment as described above, the following tests were performed after the intervention period ended: After the intervention period ended, cells were collected, and the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were measured. The results are shown below. Figures 9-11 , Figure 9 This refers to the concentration of IL-1β in the cell lysate. Figure 10 This refers to the concentration of IL-6 in the cell lysate. Figure 11 This represents the concentration of TNF-α in the cell lysate.

[0055] ① Sample preparation for inflammatory factor detection: Cell homogenate preparation: Adherent cells were gently washed with cold PBS, then digested with trypsin, and collected by centrifugation at 1000 rpm for 5 min. The collected cells were washed three times with cold PBS. Every 10... 6 Each cell was resuspended in 150-200 μL of PBS and lysed using reverse sonication. The extract was centrifuged at 1500 rpm for 10 min at 2-8°C. The supernatant was aliquoted and stored at -80°C for the detection of intracellular inflammatory factors.

[0056] ② Detection of inflammatory factors: The levels of IL-1β, IL-6, and TNF-α in cell lysates were detected using enzyme-linked immunosorbent assay (ELISA). The detection process was strictly performed according to the instructions of the corresponding ELISA kits (provided by Wuhan Elite Biotechnology Co., Ltd.). The absorbance values ​​were read at the specified wavelength (450 nm) using a microplate reader, and the concentrations of each inflammatory factor were calculated based on the standard curve provided by the kit.

[0057] from Figures 9-11 The biochemical results showed that, compared with the normal control group (NC), the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in the cell lysate of HepG2 cells induced by high glucose and high lipid were significantly increased (P<0.01). This indicates that high glucose and high lipid metabolic stress successfully activated the inflammatory response of cells, simulating the chronic inflammatory imbalance under the state of glucose and lipid metabolism disorder.

[0058] After intervention with different concentrations of corn cob xylooligosaccharides (125 μg / mL, 250 μg / mL, and 500 μg / mL), the levels of the aforementioned inflammatory factors inside and outside XL, XM, and XH cells all showed a significant dose-dependent decreasing trend. Among them, the expression and release of inflammatory factors in XL (250 μg / mL) and XH (500 μg / mL) were significantly lower than those in the model group (P<0.01), and the levels of inflammatory factors in the low-dose group also decreased compared with the model group.

[0059] The above results indicate that xylooligosaccharides from corn cobs can significantly inhibit the synthesis and secretion of inflammatory factors in HepG2 cells induced by high glucose and high lipid levels in a dose-dependent manner, effectively alleviating the inflammatory activation state at the cellular level, and providing reliable cellular experimental evidence for improving glucose and lipid metabolism disorders by regulating inflammatory responses.

[0060] (V) Effects of xylooligosaccharides from corn cobs on oxidative stress By detecting the levels of intracellular oxidative stress-related markers induced by high glucose and high lipid levels in HepG2 cells, this study assessed the regulatory effect of corn cob xylooligosaccharide (XO) on hepatocyte oxidative stress, providing experimental evidence for its ability to improve oxidative damage caused by glucose and lipid metabolism disorders.

[0061] Using the same intervention experiment as described above, after the intervention period ended, cell samples were collected for the detection of oxidative stress indicators: Cell culture supernatant collection: Carefully aspirate the culture supernatant, centrifuge at 3000 rpm for 15 min at 4℃ to remove cell debris, collect the clear supernatant, aliquot and store at -80℃ for later use in detecting the contents of glutathione peroxidase (GSH-PX), malondialdehyde (MDA) and superoxide dismutase (SOD).

[0062] Cell sample (cell homogenate) preparation: Digest cells with trypsin (wash with 0.5-1 mL isotonic PBS after digestion), then transfer the cell suspension to another centrifuge tube, centrifuge at 1000 rpm for 10 minutes, discard the supernatant, and keep the cell pellet; wash 1-2 more times with isotonic PBS, centrifuge at 1000 rpm for 10 minutes, discard the supernatant, and keep the cell pellet; add 0.2-0.3 mL of homogenization medium (physiological saline) to the cell pellet, and gently mix the cell solution to make it homogeneous. Take a small amount for cell counting. If the cell count is more than 1 million, lyse the cells using lysis buffer (Triton X-100, 1-2% concentration, 0.1 mL), lyse on ice for 30-40 minutes. The lysed liquid can be directly measured without centrifugation.

[0063] Glutathione peroxidase (GSH-PX) content determination: A kit from Nanjing Jiancheng Bioengineering Institute was used to determine the content using a colorimetric method. The absorbance was measured at a wavelength of 412 nm, and the GSH-PX content was calculated. Results are shown below. Figure 12 and Figure 13 , Figure 12 The content of GSH-PX in cell culture supernatant. Figure 13 This represents the GSH-PX content in cell homogenate.

[0064] Malondialdehyde (MDA) content determination: The MDA content was determined using a reagent kit from Nanjing Jiancheng Bioengineering Institute according to the thiobarbituric acid (TBA) method. The absorbance was measured at 532 nm, and the MDA content, reflecting the degree of lipid peroxidation, was calculated. Results are shown below. Figure 14 and Figure 15 , Figure 14 This refers to the MDA content in the cell culture supernatant. Figure 15 This represents the MDA content in the cell homogenate.

[0065] Superoxide dismutase (SOD) activity assay: The assay was performed using a kit from Nanjing Jiancheng Bioengineering Institute according to the WST-1 method. Absorbance was measured at 450 nm, and SOD activity, reflecting the tissue's antioxidant capacity, was calculated. Results are shown below. Figure 16 and Figure 17 , Figure 16 The SOD content in the cell culture supernatant. Figure 17 This represents the SOD content in the cell homogenate.

[0066] from Figures 12-17 The results of oxidative stress index detection showed that, compared with the normal control group (NC), the malondialdehyde (MDA) content in HepG2 cells induced by high glucose and high lipid was significantly increased (P<0.05), and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were significantly decreased (P<0.01), indicating that the cells were in a state of obvious oxidative stress, lipid peroxidation damage was aggravated, and the function of the endogenous antioxidant defense system was impaired.

[0067] After intervention with different concentrations of corn cob xylooligosaccharides (125 μg / mL, 250 μg / mL, and 500 μg / mL), compared with the model group, the intracellular MDA content of XL, XM, and XH cells decreased in a dose-dependent manner, while the activities of SOD and GSH-PX increased in a dose-dependent manner. Among them, the medium-dose group (XM) and the high-dose group (XH) showed particularly significant effects in improving oxidative stress imbalance (P<0.05), with the high-dose group exhibiting the most prominent reversal effect.

[0068] The above results indicate that corn cob xylooligosaccharides can dose-dependently inhibit the production of MDA, a lipid peroxidation product, in HepG2 cells induced by high sugar and high lipid levels. At the same time, they can effectively enhance the antioxidant activity of SOD and GSH-PX, significantly reduce cellular oxidative damage, repair the function of the endogenous antioxidant defense system, improve cellular redox homeostasis, and exert a protective effect against high sugar and high lipid-related oxidative stress.

[0069] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. Application of corn cob xylooligosaccharides in the preparation of drugs that regulate glucose and lipid metabolism disorders.

2. The application according to claim 1, characterized in that, The symptoms caused by the glucose and lipid metabolism disorders include one or more of the following: glucose metabolism disorders, lipid metabolism disorders, inflammatory responses, and oxidative stress damage.

3. The application according to claim 1, characterized in that, The concentration of the corn cob xylooligosaccharide used is 125~500 μg / mL.

4. Application of corn cob xylooligosaccharides in the preparation of drugs that improve glucose metabolism disorders.

5. Application of corn cob xylooligosaccharides in the preparation of drugs that reduce triglyceride and / or free fatty acid content.

6. Application of corn cob xylooligosaccharides in the preparation of drugs that inhibit inflammatory responses caused by glucose and lipid metabolism disorders.

7. Application of corn cob xylooligosaccharides in the preparation of drugs that alleviate oxidative stress damage caused by glucose and lipid metabolism disorders.

8. A pharmaceutical composition for treating disorders of glucose and lipid metabolism, characterized in that, This includes xylooligosaccharides from corn cobs and pharmaceutically acceptable carriers.

9. The pharmaceutical composition according to claim 8, characterized in that, The dosage form of the pharmaceutical composition includes oral or injectable formulations.

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