Composition with antioxidant function and application

Abstract

The invention discloses a composition with an antioxidant function and application, and belongs to the technical field of antioxidants. The composition with the antioxidant function comprises a propolis extract, curcumin and nervonic acid, the weight ratio of the propolis extract to the curcumin to the nervonic acid is (2-6): (2-5): (1-5). The three substances are compounded and cooperated, so that the content of total superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) can be increased, the content of malonaldehyde (MDA), protein carbonyl, glutamic-pyruvic transaminase (ALT) and glutamic oxalacetic transaminase (AST) can be reduced, the anti-oxidation effect is achieved, and the safety is good.

Description

Technical Field This invention relates to the field of antioxidant technology, and more specifically to a composition with antioxidant function and its application. Background Technology

[0002] Oxidative stress is a major factor contributing to material aging, food spoilage, and various chronic diseases in humans, such as cardiovascular disease, neurodegenerative diseases, and cancer. Antioxidants, as substances that can scavenge free radicals and inhibit oxidation reactions, are widely used in oils, foods, cosmetics, and pharmaceutical products to slow down the oxidation process, maintain product stability, extend shelf life, and exert their health-promoting effects.

[0003] Currently, commonly used antioxidants are mainly divided into two categories: synthetic antioxidants and natural antioxidants. Synthetic antioxidants, such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and tert-butylhydroquinone (TBHQ), while possessing clear free radical scavenging effects, suffer from potential toxicity and low bioavailability. Therefore, developing safe and efficient natural antioxidants has become a current research hotspot. Common natural antioxidants include vitamin C, vitamin E, polyphenols, and carotenoids. Although these natural ingredients have high safety profiles, their practical application still faces many technical bottlenecks, such as the limited effectiveness of single-ingredient antioxidants and the potential for adverse effects (vitamin E, as a classic antioxidant supplement, can cause side effects with long-term use).

[0004] Therefore, it is essential to develop a composition that is safe and has good antioxidant effects. Summary of the Invention

[0005] In order to overcome the above-mentioned shortcomings of the prior art, the purpose of this invention is to provide a composition and application with antioxidant function, so as to solve the problem that the antioxidant effect and safety of existing antioxidants cannot be satisfied at the same time.

[0006] The technical solution of the present invention to solve the above-mentioned technical problems is as follows: a composition with antioxidant function is provided, comprising propolis extract, curcumin and nervonic acid; the weight ratio of propolis extract, curcumin and nervonic acid is 2-6:2-5:1-5.

[0007] The beneficial effects of this invention are as follows: the propolis extract is rich in flavonoids, phenolic acids, and other components, which can provide hydrogen atoms or electrons to neutralize various reactive oxygen species and reactive nitrogen species in a broad-spectrum and highly efficient manner, such as hydroxyl radicals (·OH) and superoxide anions (·O2). - (e.g., peroxynitrite, etc.) act as the first line of defense.

[0008] Curcumin's enone structure enables it to effectively quench singlet oxygen and inhibit the propagation of lipid peroxidation chain reactions. More importantly, curcumin can downregulate the expression of prooxidases and activate the cell's own antioxidant defense system at its source.

[0009] Nervonic acid, a core component of the myelin sheath and cell membranes of brain nerve cells, is not itself a potent direct antioxidant. However, it can synergistically exert excellent antioxidant effects with propolis extract and curcumin, specifically: Propolis extract rapidly eliminates various free radicals present in the system, reducing overall oxidative stress. After propolis extract eliminates most free radicals, curcumin continues to block stubborn chain reactions, enhancing the long-term antioxidant capacity of the body's cells. Nerve acid propolis extract and curcumin provide a more stable and protected platform, which, by strengthening cell membranes and reducing the "attack surface" of oxidative damage, allows propolis extract and curcumin to act more efficiently on remaining oxidative threats, while simultaneously providing repair materials for damaged nerve membrane structures. The combined use of these three components achieves a comprehensive defense from direct elimination—blocking propagation and systemic enhancement—to structural protection and repair, with effects far exceeding the simple sum of the functions of individual components.

[0010] Based on the above technical solution, the present invention can be further improved as follows: Furthermore, the weight ratio of propolis extract, curcumin, and nervonic acid is 4:2.7:3.

[0011] Furthermore, the propolis extract was prepared by the following method: Propolis was extracted with ethanol, followed by ultrasonic-assisted extraction, and finally the extract was concentrated to obtain propolis extract.

[0012] Furthermore, the volume concentration of ethanol is 90-95%, and the extraction time is 10-15 hours; preferably, the volume concentration of ethanol is 95%, and the extraction time is 12 hours.

[0013] Furthermore, the volume ratio of propolis to ethanol is 1:8-15; preferably, the volume ratio of propolis to ethanol is 1:10.

[0014] Furthermore, the ultrasonic time for ultrasonic-assisted extraction is 15-25 min, the ultrasonic power is 180-220 W, and the ultrasonic temperature is 25-35℃; preferably, the ultrasonic time is 20 min, the ultrasonic power is 200 W, and the ultrasonic temperature is 30℃.

[0015] The above-mentioned antioxidant compositions are used in the preparation of drugs with antioxidant functions.

[0016] Furthermore, the aforementioned antioxidant functions include increasing the levels of superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH), while decreasing the levels of malondialdehyde (MDA), protein carbonyl groups, alanine aminotransferase (ALT), and aspartate aminotransferase (AST).

[0017] A drug with antioxidant function, comprising the above-described composition and pharmaceutically acceptable excipients.

[0018] The present invention has the following beneficial effects: The antioxidant composition provided by this invention comprises three traditional Chinese medicine ingredients: propolis extract, curcumin, and nervonic acid. Propolis is rich in flavonoids and phenolic acids, exhibiting excellent antioxidant activity. Curcumin, as a plant used in both medicine and food, possesses antioxidant, blood pressure-lowering, immunity-enhancing, and cardiovascular and digestive system-wide therapeutic effects. Nervonic acid, an unsaturated fatty acid, promotes nerve cell repair, improves cognitive function, regulates lipid metabolism, and has various physiological activities affecting the nervous and cardiovascular systems. The synergistic effect of the three substances can increase the levels of total superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH), while decreasing the levels of malondialdehyde (MDA), protein carbonyl groups, alanine aminotransferase (ALT), and aspartate aminotransferase (AST), thus achieving antioxidant effects with good safety. Attached Figure Description

[0019] Figure 1 The images show histopathological sections of mouse liver tissue from each treatment group; green arrows indicate watery degeneration; yellow arrows indicate hepatocellular necrosis; purple arrows indicate fatty degeneration; and orange arrows indicate disordered arrangement of hepatic cords. Detailed Implementation

[0020] The sources of the materials, reagents, and animals used in this invention are as follows: 1. Animals KM mice, all male, with a fasting weight of 20±2g, were provided by Chengdu Dashuo Experimental Animal Co., Ltd.

[0021] 2. Materials and Reagents Propolis was purchased from Sichuan Jianshengtang Agricultural Development Co., Ltd.; curcumin from Guosheng Biotechnology Co., Ltd.; nervonic acid from Yunnan Beimu Biotechnology Co., Ltd.; anhydrous ethanol from Chongqing Wansheng Chuandong Chemical Co., Ltd.; polyethylene glycol 400 from Chengdu Jinshan Chemical Reagent Co., Ltd.; catalase assay kit (CAT, catalog number A007-1-1) from Nanjing Jiancheng Technology Co., Ltd.; malondialdehyde assay kit (MDA, catalog number A003-1) from Nanjing Jiancheng Technology Co., Ltd.; total superoxide dismutase assay kit (T-SO4) was purchased from Nanjing Jiancheng Technology Co., Ltd. D (item number A001-1) was purchased from Nanjing Jiancheng Technology Co., Ltd.; Protein carbonyl content assay kit (item number A087-1) was purchased from Nanjing Jiancheng Technology Co., Ltd.; Micro-reduced glutathione assay kit (GSH, item number A006-2-1) was purchased from Nanjing Jiancheng Technology Co., Ltd.; Alanine aminotransferase (ALT, item number 105-000442-00) was purchased from Mindray Bio-Medical Electronics Co., Ltd.; Aspartate aminotransferase (AST, item number 105-000443-00) was purchased from Mindray Bio-Medical Electronics Co., Ltd.

[0022] The examples given below are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified, conditions in the examples are performed under standard conditions or as recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0023] Example 1: An antioxidant composition comprising propolis extract, curcumin, and nervonic acid; wherein the weight ratio of propolis extract, curcumin, and nervonic acid is 2:2:1.

[0024] The above-mentioned propolis extract was prepared by the following method: the propolis raw material was extracted with 10 times its volume of 95% ethanol for 12 hours, and then extracted with ultrasound-assisted extraction. After extraction three times, the supernatants were combined, concentrated and evaporated to constant weight to obtain propolis ethanol extract; wherein, the ultrasound time for ultrasound-assisted extraction was 15 minutes, the ultrasound power was 220W, and the ultrasound temperature was 35℃.

[0025] Example 2: An antioxidant composition comprising propolis extract, curcumin, and nervonic acid; wherein the weight ratio of propolis extract, curcumin, and nervonic acid is 6:5:5.

[0026] The above-mentioned propolis extract was prepared by the following method: the propolis raw material was extracted with 10 times its volume of 95% ethanol for 12 hours, and then extracted with ultrasound-assisted extraction. After extraction three times, the supernatants were combined, concentrated and evaporated to constant weight to obtain propolis ethanol extract; wherein, the ultrasound time for ultrasound-assisted extraction was 25 minutes, the ultrasound power was 180W, and the ultrasound temperature was 25℃.

[0027] Example 3: An antioxidant composition comprising propolis extract, curcumin, and nervonic acid; wherein the weight ratio of propolis extract, curcumin, and nervonic acid is 4:2.7:3.

[0028] The above-mentioned propolis extract was prepared by the following method: the propolis raw material was extracted with 10 times its volume of 95% ethanol for 12 hours, and then extracted with ultrasound assistance. After extraction three times, the supernatants were combined and concentrated and evaporated to constant weight to obtain propolis ethanol extract; wherein, the ultrasound time during ultrasound-assisted extraction was 20 minutes, the ultrasound power was 200W, and the ultrasound temperature was 30℃.

[0029] Test case The composition of the present invention with antioxidant function was used for efficacy testing, and the specific process is as follows: 1. Animal grouping and treatment Seventy mice with an average weight of 20±2g were randomly divided into four groups according to the groupings in Table 1: a blank control group, a model control group, a positive control group, and treatment groups (including propolis + curcumin group, propolis + nervonic acid group, curcumin + nervonic acid group, and propolis + curcumin + nervonic acid group). Each group of animals was given distilled water and normal feed in the morning. The treatment groups were administered the respective combination of ingredients by gavage every morning (the weight ratio of propolis extract, curcumin, and nervonic acid in the treatment groups was 4:2.7:3; for example, the weight ratio of propolis extract to curcumin in the propolis + curcumin group was 4:2.7, in the propolis + nervonic acid group it was 4:3, in the curcumin + nervonic acid group it was 2.7:3, and in the propolis + curcumin + nervonic acid group it was 4:2.7:3). The dosages are shown in Table 1.

[0030] Table 1. Animal grouping and treatment 2. Indicator Testing Thirty days after treatment, mice in each group were fasted for 16 hours (overnight) after the last gavage. Then, they were given 12 mL / kg BW of 50% ethanol by gavage. Six hours later, liver tissue samples were collected (the blank control group was not treated and was collected without fasting). The contents of CAT, MDA, T-SOD, GSH, protein carbonyl, ALT, and AST in liver tissue were measured. After fixation with 10% formaldehyde solution, the liver tissue was routinely dehydrated, embedded in paraffin, sectioned, stained with hematoxylin and eosin (HE), and the pathological changes of mouse liver tissue were observed under an optical microscope.

[0031] The determination methods for CAT, MDA, T-SOD, GSH, protein carbonyl, ALT, and AST should be performed according to the kit instructions.

[0032] 3. Data Processing Experimental data are expressed as x±s, and statistical analysis of variance was performed using SPSS 10.0 statistical software.

[0033] 4. Results and Analysis (1) Results of biochemical index determination of mouse liver tissue The results of the biochemical index determination of mouse liver tissue are shown in Table 2: Table 2 Effects of experimental treatments on biochemical parameters of mouse liver tissue Note: Compared with the normal group, ####P<0.0001; compared with the model group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0034] Table 2 shows that, compared with the blank control group, the levels of CAT, T-SOD, and GSH in the model control group were significantly decreased (p<0.0001), while the levels of MDA and protein carbonyl groups were significantly increased (p<0.0001). Compared with the model control group, the CAT level in each treatment group was increased, with a significant increase in the propolis + curcumin group (p<0.01), a significant increase in the curcumin + nervonic acid group (p<0.001), and an extremely significant increase in the propolis + nervonic acid group and the propolis + curcumin + nervonic acid group (p<0.0001). The MDA level in the curcumin + nervonic acid group was significantly decreased (p<0.01), a significant decrease in the propolis + curcumin group and the propolis + nervonic acid group (p<0.001), and an extremely significant decrease in the propolis + curcumin + nervonic acid group (p<0.0001). The T-SOD level was significantly increased in the propolis + nervonic acid group (p<0.001), and extremely significantly increased in the propolis + curcumin + nervonic acid group (p<0.0001). The GSH level was significantly increased in the propolis + nervonic acid group (p<0.05), significantly increased in the propolis + curcumin group (p<0.01), and extremely significantly increased in the propolis + curcumin + nervonic acid group (p<0.001). The levels were significantly decreased in the propolis + curcumin group, curcumin + nervonic acid group, and propolis + nervonic acid group (p<0.001), and extremely significantly decreased in the propolis + curcumin + nervonic acid group (p<0.0001). Therefore, the antioxidant capacity of the propolis + curcumin + nervonic acid group was superior to that of the other treatment groups.

[0035] (2) Results of mouse liver injury assay For details of the mouse liver injury assay results, please refer to [link / reference]. Figure 1 And Table 3: Table 3 Effects of experimental treatments on liver injury in mice Note: Compared with the normal group, ####P<0.0001; compared with the model group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0036] As shown in Table 3, compared with the blank control group, the ALT and AST levels in the model control group were significantly increased (P<0.0001). Compared with the model control group, the ALT and AST levels in mice in each treatment group were decreased to varying degrees. Among them, the ALT levels in the curcumin + nervonic acid group and the propolis + nervonic acid group were significantly decreased (P<0.01), the propolis + curcumin group was extremely significantly decreased (P<0.001), and the propolis + curcumin + nervonic acid group was extremely significantly decreased (P<0.001). The AST levels in the propolis + curcumin group were significantly decreased (P<0.05), and the levels in the curcumin + nervonic acid group, the propolis + nervonic acid group, and the propolis + curcumin + nervonic acid compound group were significantly decreased (P<0.01).

[0037] Depend on Figure 1 It was found that in the blank control group, the liver lobule structure of mice was clear, with no obvious lesions. Hepatocytes were arranged radially around the central vein, and the sinusoidal structure was regular. In the model control group, hepatocytes showed fatty degeneration, fat vacuoles, inflammatory cell infiltration, enlarged intercellular spaces, and disordered arrangement. Compared with the model control group, hepatocyte necrosis and inflammatory cell infiltration were improved in all drug-treated groups. The arrangement of hepatocytes was more complete and orderly, the number of necrotic hepatocytes was reduced, and the number of cell nuclei was increased. Among them, the propolis + curcumin + nervonic acid group showed more significant improvement in liver tissue than the other drug-treated groups.

[0038] (3) Results of mouse inflammatory factor assay The results of the mouse inflammatory factor assay are shown in Table 4: Table 4. Effects of experimental treatments on the results of inflammatory factors in mice. Note: Compared with the normal group, ####P<0.0001; compared with the model group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0039] As shown in Table 4, compared with the blank control group, the levels of TNF, IL-1β, and IL-6 in the model control group were significantly increased (P<0.0001). Compared with the model control group, the levels of TNF, IL-1β, and IL-6 in mice in each treatment group were decreased to varying degrees. Among them, the TNF levels in the propolis + curcumin group, curcumin + nervonic acid group, propolis + nervonic acid group, and propolis + curcumin + nervonic acid group were extremely significantly decreased (P<0.001). The IL-1β levels in the propolis + curcumin group, curcumin + nervonic acid group, and propolis + nervonic acid group were significantly decreased (P<0.05), and the IL-1β levels in the propolis + curcumin + nervonic acid group were extremely significantly decreased (P<0.001). The IL-6 levels in the propolis + nervonic acid group were significantly decreased (P<0.01), the IL-6 levels in the curcumin + nervonic acid group were significantly decreased (P<0.001), and the IL-6 levels in the propolis + curcumin group and the propolis + curcumin + nervonic acid group were extremely significantly decreased (P<0.001). Therefore, the anti-inflammatory effect of the propolis + curcumin + nervonic acid group was better than that of other drug administration groups.

[0040] 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, improvements, etc., 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 having antioxidant function, characterized in that, It includes propolis extract, curcumin and nervonic acid; the weight ratio of the propolis extract, curcumin and nervonic acid is 2-6:2-5:1-5.

2. The composition with antioxidant function according to claim 1, characterized in that, The weight ratio of the propolis extract, curcumin, and nervonic acid is 4:2.7:

3.

3. The composition with antioxidant function according to claim 1, characterized in that, Propolis extract is prepared by the following method: Propolis was extracted with ethanol, followed by ultrasonic-assisted extraction, and finally the extract was concentrated to obtain propolis extract.

4. The composition with antioxidant function according to claim 3, characterized in that, The volume concentration of ethanol is 90-95%, and the extraction time is 10-15 hours.

5. The composition with antioxidant function according to claim 3 or 4, characterized in that, The volume ratio of propolis to ethanol is 1:8-15.

6. The composition with antioxidant function according to claim 5, characterized in that, The volume ratio of propolis to ethanol is 1:

10.

7. The composition with antioxidant function according to claim 3, characterized in that, The ultrasonic extraction time is 15-25 min, the ultrasonic power is 180-220 W, and the ultrasonic temperature is 25-35℃.

8. Use of the composition with antioxidant function according to any one of claims 1-7 in the preparation of a medicament with antioxidant function.

9. The application according to claim 8, characterized in that, Its antioxidant function is to increase the levels of superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH), while decreasing the levels of malondialdehyde (MDA), protein carbonyl groups, alanine aminotransferase (ALT), and aspartate aminotransferase (AST).

10. A drug with antioxidant function, characterized in that, This includes the composition with antioxidant function as described in any one of claims 1-7, as well as pharmaceutically acceptable excipients.

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