A composition for reducing gastroesophageal reflux in an individual, and methods of making and using the same

Abstract

The present application relates to the field of food technology, and specifically discloses a composition for reducing gastroesophageal reflux in individuals, a preparation method and application thereof. The composition for reducing gastroesophageal reflux in individuals comprises beta-defensin 1, lysozyme and lactoperoxidase. The present application constructs a multi-target synergistic intervention mechanism combining physical protection, secretion regulation and tissue repair through the compounding of four core components, which can comprehensively block the occurrence and development of gastroesophageal reflux. Lactadherin constructs a physical barrier to reduce the risk of reflux dynamics from the source; MLCT precisely regulates gastric acid secretion to reduce chemical stimulation; HMO complex IgG strengthens the mucosal immune barrier and promotes damage repair. The above composition is applied to products for reducing gastroesophageal reflux in individuals, and multi-target synergistic effect is achieved, realizing a full-chain closed-loop intervention from prevention, relief to repair, and significantly improving the clinical improvement effect.

Description

Technical Field

[0001] This invention relates to the field of food technology, and in particular to a composition for reducing gastroesophageal reflux in individuals, its preparation method, and its application. Background Technology

[0002] Gastroesophageal reflux (GER) refers to a series of symptoms caused by the reflux of stomach and duodenal contents into the esophagus. Its incidence rate is as high as 50%–70% in infancy and 10%–30% in adults. The pathophysiological mechanism of GER is complex, involving multiple factors such as transient lower esophageal sphincter relaxation (TLESR), decreased esophageal clearance capacity, delayed gastric emptying, and impaired esophageal mucosal barrier function.

[0003] Currently, commonly used clinical interventions for gastroesophageal reflux disease (GERD) include pharmacological intervention, dietary thickening, combined pharmacological and dietary intervention, and surgical intervention. Medications include proton pump inhibitors (PPIs) and H2 receptor antagonists, which suppress acidity. While these drugs can alleviate GERD symptoms by reducing the acidity of refluxed material, long-term use may lead to adverse reactions such as malabsorption, gut microbiota dysbiosis, and increased risk of osteoporosis. Furthermore, these acid-suppressing drugs are ineffective against non-acidic reflux. Dietary thickening involves adding thickeners such as lactobacillus and rice starch to the diet to physically reduce reflux by increasing the viscosity of gastric contents. Thickened formula foods are currently the standard nutritional intervention for GERD management, but their mechanism of action is singular, relying solely on physical thickening and failing to address fundamental issues such as esophageal mucosal repair, inflammation control, and improved gastric emptying. While combining acid-suppressing drugs with thickened formula foods (i.e., combined pharmacological and dietary intervention) can partially synergistically improve symptoms, it suffers from poor adherence to multiple medications and high costs. In surgical interventions, fundoplication is suitable for severe gastroesophageal reflux, but it is highly invasive, expensive, and carries the risk of postoperative complications.

[0004] Existing thickening-type formula foods only target the single physical barrier mechanism, failing to address multiple aspects simultaneously, such as esophageal mucosal repair, immune inflammation regulation, and improved gastric emptying. Therefore, there is an urgent need to develop an intervention composition that is synergistically effective through multiple mechanisms, safe, efficient, and suitable for long-term use. Summary of the Invention

[0005] To address the aforementioned problems, this invention provides a composition for reducing individual gastroesophageal reflux, its preparation method, and its application. Through the design of the composition, a triple synergistic mechanism is employed, involving β-defensin 1 (hBD1) regulating gene expression, lysozyme clearing the inducing factors, and lactoperoxidase repairing tissue structure, effectively reducing the frequency and amount of individual gastroesophageal reflux.

[0006] To achieve the above-mentioned objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a composition for reducing individual gastroesophageal reflux, comprising β-defensin 1, lysozyme, and lactoperoxidase.

[0007] Compared to existing technologies, the composition provided by this invention for reducing individual gastroesophageal reflux utilizes β-defensin 1, which targets and silences key genes in inflammatory pathways (such as TRAF6 and IRAK1), blocking abnormal immune responses at the gene level, improving esophageal sphincter function, and restoring the serum pepsinogen I / II ratio (PG I / II ratio), reflecting the overall health status of the gastroesophageal axis. Lysozyme, acting as an enzymatic barrier, hydrolyzes pathogenic bacteria in the stomach, reducing lower esophageal sphincter dysfunction and chronic inflammation induced by infection, thereby reducing myeloperoxidase (MPO) activity and malondialdehyde (MDA) content, alleviating acute inflammatory infiltration and oxidative stress damage. Lactoperoxidase promotes esophageal epithelial cell migration and proliferation, accelerates the healing of damaged mucosa, and directly reduces the esophageal mucosal damage index. The combination of β-defensin 1, lysozyme, and lactoperoxidase, through a synergistic mechanism of source blocking, enzymatic clearance, and structural repair, comprehensively intervenes in multiple aspects of gastroesophageal reflux.

[0008] Preferably, the composition comprises the following components in parts by weight: β-defensin 1 1 × 10 6 1.5 x 10 portions 7 5×10 portions of lysozyme 4 5 x 10 5 0.01 to 0.2 parts of lactoperoxidase.

[0009] More preferably, the composition comprises the following components in parts by weight: β-defensin 1 3 × 10 6 1.2 x 10 portions 7 8×10 portions of lysozyme 4 4 x 10 servings 5 0.03 to 0.15 parts of lactoperoxidase.

[0010] More preferably, the composition comprises the following components in parts by weight: β-defensin 1 4 × 10 6 1 x 10 7 1 serving, lysozyme 1×10 5 3 x 10 servings 5 0.05 to 0.12 parts of lactoperoxidase.

[0011] More preferably, the composition comprises the following components in parts by weight: β-defensin 1.5 × 10 6 8 x 10 servings 6 Part, lysozyme 1.2×105 Servings ~ 2 x 10 5 0.06 to 0.1 parts of lactoperoxidase.

[0012] In a second aspect, the present invention provides a method for preparing the composition for reducing individual gastroesophageal reflux, comprising the following steps: β-defensin 1, lysozyme, and lactoperoxidase are mixed evenly to obtain a composition for reducing individual gastroesophageal reflux.

[0013] Thirdly, the present invention provides a product comprising the aforementioned composition for reducing individual gastroesophageal reflux.

[0014] Preferably, the product includes foods for special medical purposes.

[0015] The present invention has the following beneficial effects: This invention provides a composition for reducing gastroesophageal reflux disease (GERD) in individuals, comprising β-defensin 1, lysozyme, and lactoperoxidase. Compared to the local relief effects of single-component or simple compound formulations in existing technologies, this invention constructs a three-dimensional synergistic mechanism of source blocking, enzymatic clearance, and structural repair through specific ingredient combinations. The three active ingredients, through a precise cascade reaction, form a complete pathological intervention loop: β-defensin 1 regulates gene expression, lysozyme clears the inducing factor, and lactoperoxidase repairs tissue structure. This achieves multi-target synergistic effects and systematic intervention throughout the entire process of GERD occurrence and development.

[0016] Experiments have confirmed that the composition of this invention can significantly reduce the damage index of the esophageal mucosa, indicating that it effectively alleviates the physical and chemical damage to the esophageal epithelium caused by reflux at the macroscopic level. Simultaneously, the composition can significantly downregulate myeloperoxidase activity and malondialdehyde content, demonstrating that it effectively inhibits neutrophil infiltration and lipid peroxidation at the microscopic level, greatly alleviating local inflammatory stress. This composition not only repairs the morphological structure of the gastroesophageal tract but also significantly increases the serum pepsinogen I / II ratio. The PG I / II ratio is a key biomarker reflecting the secretory function of the chief cells of the gastric fundic gland mucosa; its significant increase confirms that this invention can substantially improve glandular atrophy and restore the normal secretory and defensive functions of the gastric mucosa, rather than merely providing temporary symptom relief. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0018] Unless otherwise specified, all materials used in the embodiments of this invention can be obtained through commercial channels or prepared by conventional methods in the art.

[0019] Example 1 This embodiment provides a composition for reducing individual gastroesophageal reflux, comprising the following components in parts by weight: β-defensin 1 6.4 g, lysozyme 150 mg and lactoperoxidase 0.075 μg.

[0020] The method for preparing the above-mentioned composition for reducing individual gastroesophageal reflux includes the following steps: Weigh each component according to the design ratio, and mix the components evenly in a dry mixer to obtain a composition for reducing individual gastroesophageal reflux.

[0021] Example 2 This embodiment provides a composition for reducing individual gastroesophageal reflux, comprising the following components in parts by weight: β-defensin 1 3g, lysozyme 300mg and lactoperoxidase 0.05μg.

[0022] The preparation method of the above-mentioned composition for reducing individual gastroesophageal reflux is the same as that in Example 1, and will not be repeated here.

[0023] Example 3 This embodiment provides a composition for reducing individual gastroesophageal reflux, comprising the following components in parts by weight: 10g of β-defensin 1, 80mg of lysozyme, and 0.12μg of lactoperoxidase.

[0024] The preparation method of the above-mentioned composition for reducing individual gastroesophageal reflux is the same as that in Example 1, and will not be repeated here.

[0025] Comparative Example 1 This comparative example provides a composition for reducing individual gastroesophageal reflux, with components and proportions similar to those in Example 1, except that β-defensin 1 is replaced with an equal mass of α-defensin 1. Specifically, the composition comprises the following components in parts by mass: 6.4 g of α-defensin 1, 150 mg of lysozyme, and 0.075 μg of lactoperoxidase.

[0026] The preparation method of the above-mentioned composition for reducing individual gastroesophageal reflux is the same as that in Example 1, and will not be repeated here.

[0027] Animal experiments verified 1. Laboratory animals and grouping Healthy adult SD rats were selected for the experiment. They were provided by Spf (Suzhou) Biotechnology Co., Ltd. (China), with the certificate number SCXK (Su) 2022-0006, and their body weights were 200 g to 300 g. The rats were required to be adaptively fed in an environment with constant temperature (22°C to 25°C) and constant humidity (50% to 60%) for 1 week, with free access to food and water, to ensure good physical condition. A relaxant (such as a vasodilator like nitroglycerin or a specific neurotransmitter regulator) was locally injected near the lower esophageal sphincter, and the sphincter relaxation was observed. When the LES pressure dropped to ≤5 mmHg, it indicated that the sphincter had been fully relaxed and reached the relaxed state required for the experiment.

[0028] The rats were fasted for 24 hours before surgery without water restriction to reduce the interference of gastric contents on the surgery. After anesthetizing the rats with isoflurane inhalation anesthesia or intraperitoneal injection of sodium pentobarbital (the dose was adjusted according to the rat body weight), the rats were fixed in the supine position on the operating table, and their four limbs were fixed with clips to keep the airway unobstructed. The skin and subcutaneous tissue were incised along the midline of the xiphoid process in the upper abdomen, and the abdominal cavity was entered layer by layer. The incision length was about 1 cm to 2 cm to expose the stomach. The hepatogastric ligament was transected, and the esophagus was dissected, taking care to preserve the vagus nerve. Incisions of 1 cm to 1.5 cm were made along the long axis at the esophagogastric junction and the free edge of the duodenum 1 cm from the pylorus respectively for esophago-duodenal side anastomosis. A 3 / 0 suture needle with thread was used for suturing, and the entire stomach was retained. Before closing the abdomen, 10 mL to 20 mL of 0.9% sodium chloride injection was poured into the abdominal cavity to wash the abdominal cavity, and then the abdominal wall was sutured layer by layer.

[0029] The rats with successful model establishment were randomly divided into 11 groups, with 12 rats in each group. The grouping is shown in Table 1. The rats in each group were continuously gavaged for 14 days, once a day. The concentration of the composition in the dosing groups G2 to G17 was 10 mg / kg, and the specific dosage for each gavage was 2.5 mL. 24 hours after the last gavage, the rats were anesthetized and sacrificed, and esophageal tissues were taken for mucosal injury index scoring, and the activities of myeloperoxidase (MPO) in esophageal tissues, the ratio of serum pepsinogen I / II (PG I / II), and the content of malondialdehyde (MDA) in esophageal tissues were detected.

[0030] Table 1 Experimental grouping design

[0031] 2. Experimental results (1) Esophageal mucosal injury index After the esophagus was incised, immediate scoring was performed according to the morphology, length, and scope of mucosal injury. The scoring criteria are shown in Table 2, and the scoring results are shown in Table 3. The higher the score, the more severe the injury.

[0032] Table 2 Scoring criteria for esophageal mucosal injury index

[0033] Table 3. Esophageal Mucosal Injury Index Scoring Results

[0034] In Table 3, # indicates a significant difference compared to G8, p < 0.05; ## indicates a significant difference compared to G8, p < 0.01; and ### indicates a significant difference compared to G8, p < 0.001.

[0035] As shown in the table above, the damage index scores of the single-use groups (G2-G4) and the paired-use groups (G5-G7) ranged from 2.3 to 3.9, indicating that although there was some improvement compared to the model group, significant inflammatory infiltration or epithelial hyperplasia still existed in the esophageal mucosa. The combined group of the three drugs reduced the index to 1.1, which was significantly lower than all other groups (p < 0.01). Furthermore, the esophageal mucosal damage index scores of G8-G10 were significantly lower than those of G11.

[0036] (2) Peroxidase activity in esophageal tissue MPO is a marker enzyme for neutrophil activation, and its activity directly reflects the degree of neutrophil infiltration in esophageal mucosa. In gastroesophageal reflux disease (GERD), refluxed material stimulates the esophageal mucosa, triggering neutrophil recruitment and activation, leading to tissue damage. Higher MPO activity indicates more severe inflammation. Esophageal tissue homogenates were analyzed using an MPO detection kit (spectrophotometry), and the results are expressed as U / g tissue, as shown in Table 4.

[0037] Table 4 Peroxidase activity in esophageal tissue

[0038] In Table 4, # indicates a significant difference compared to G8, p < 0.05; ## indicates a significant difference compared to G8, p < 0.01; and ### indicates a significant difference compared to G8, p < 0.001.

[0039] As shown in the table above, compared with the model control group, the esophageal MPO activity in the embodiment group of the present invention was significantly reduced, with a reduction rate of up to 74.1%. Moreover, the MPO activity using β-defensin 1 (G8~G10) was even lower than that using α-defensin 1 (G11). This indicates that the composition of the present invention can effectively inhibit the recruitment and activation of neutrophils to the esophageal mucosa, thereby alleviating reflux-induced acute inflammatory damage.

[0040] (3) Serum pepsinogen I / II ratio Pepsinogen I and II are secreted by the chief cells of the stomach. The PG I / II ratio changes when the gastric mucosa undergoes atrophy, intestinal metaplasia, or inflammation. In GERD patients, chronic reflux can lead to esophageal mucosal damage and gastric dysfunction. The PG I / II ratio is a non-invasive indicator for assessing the functional status of the gastric mucosa and the severity of esophageal damage. Serum was collected and PG I and PG II concentrations were measured using ELISA. The PG I / PG II ratio was calculated, and the results are shown in Table 5.

[0041] Table 5. Serum pepsinogen I / II ratio results

[0042] In Table 5, # indicates a significant difference compared to G8, p < 0.05; ## indicates a significant difference compared to G8, p < 0.01; and ### indicates a significant difference compared to G8, p < 0.001.

[0043] As can be seen from the table above, compared with the model control group, the serum PG I / II ratio of the embodiment group of the present invention increased significantly, from 4.5 to above 7.0, and returned to the normal range (≥7.0). Among them, the G8 group reached the best, indicating that the composition of the present invention can effectively improve the gastric mucosal function and reduce the overall damage of long-term reflux to the gastroesophageal system.

[0044] (4) Malondialdehyde content in esophageal tissue MDA is the end product of lipid peroxidation, reflecting the degree of attack of free radicals on the esophageal mucosal epithelial cell membrane. Gastric acid and bile acids in GERD reflux can induce oxidative stress, leading to cell membrane damage. MDA content is a key indicator for assessing GERD-related oxidative damage. Esophageal tissue homogenates were analyzed using the thiobarbituric acid (TBA) method, and the results are expressed as nmol / mg protein, as shown in Table 6.

[0045] Table 6. Malondialdehyde content in esophageal tissue

[0046] In Table 6, # indicates a significant difference compared to G8, p < 0.05; ## indicates a significant difference compared to G8, p < 0.01; and ### indicates a significant difference compared to G8, p < 0.001.

[0047] As shown in the table above, compared with the model control group, the MDA content in the esophageal tissue of the present invention embodiment group was significantly reduced, with a reduction rate as high as 70.6% (from 6.8 nmol / mg in the model group to 2.0 nmol / mg), returning to the normal reference range (≤2.0 nmol / mg). This indicates that the composition of the present invention can effectively scavenge free radicals and inhibit lipid peroxidation, thereby alleviating reflux-induced oxidative stress damage and protecting the integrity of the esophageal mucosal epithelial cell membrane.

[0048] In summary, the composition provided by this invention for reducing individual gastroesophageal reflux disease (GERD) comprises: β-defensin 1, which acts as an immune regulatory hub, deeply regulating epithelial cell gene expression and enhancing the innate mucosal immune barrier; lysozyme, which functions as a scavenger, reducing the pathogenic bacterial load and exogenous inflammatory factors by hydrolyzing the cell walls of Gram-positive bacteria; and lactoperoxidase, which acts as a repair engine, promoting the regeneration and remodeling of damaged tissue structures by utilizing the antibacterial substances and oxidative environment produced by catalysis. These four components produce a super-synergistic effect in reducing GERD, comprehensively intervening in the pathological processes of GERD from its source to its end, achieving a multi-target, full-chain therapeutic effect that cannot be achieved by a single component or any combination of components.

[0049] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A composition for reducing individual gastroesophageal reflux, characterized in that: It includes β-defensin 1, lysozyme, and lactoperoxidase.

2. The composition for reducing individual gastroesophageal reflux as described in claim 1, characterized in that: The composition comprises the following components in parts by weight: β-defensin 1 1 × 10 6 1.5 x 10 portions 7 5×10 portions of lysozyme 4 5 x 10 5 0.01 to 0.2 parts of lactoperoxidase.

3. The composition for reducing individual gastroesophageal reflux as described in claim 2, characterized in that: The composition comprises the following components in parts by weight: β-defensin 1 3 × 10 6 1.2 x 10 portions 7 8×10 portions of lysozyme 4 4 x 10 servings 5 0.03 to 0.15 parts of lactoperoxidase.

4. The composition for reducing individual gastroesophageal reflux as described in claim 3, characterized in that: The composition comprises the following components in parts by weight: β-defensin 1 4 × 10 6 1 x 10 7 1 serving, 1×10 lysozyme 5 3 x 10 servings 5 0.05 to 0.12 parts of lactoperoxidase.

5. The composition for reducing individual gastroesophageal reflux as described in claim 4, characterized in that: The composition comprises the following components in parts by weight: β-defensin 1.5 × 10 6 8 x 10 servings 6 Part, lysozyme 1.2×10 5 Servings ~ 2 x 10 5 0.06 to 0.1 parts of lactoperoxidase.

6. A method for preparing the composition for reducing individual gastroesophageal reflux according to any one of claims 1 to 5, characterized in that: Includes the following steps: β-defensin 1, lysozyme, and lactoperoxidase are mixed evenly to obtain a composition for reducing individual gastroesophageal reflux.

7. A product, characterized in that: Includes the composition for reducing individual gastroesophageal reflux as described in any one of claims 1 to 5.

8. The product as described in claim 7, characterized in that: The products include foods formulated for special medical purposes.

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