Application of lycopene in preparation of product for relieving body injury caused by chronic cold exposure

Abstract

The invention relates to the technical field of medicines, in particular to application of lycopene in preparation of a product for relieving body injury caused by chronic cold exposure. The lycopene is found to be capable of relieving inflammatory response of chronic cold exposure mice, reducing oxidative stress level and improving body immune function of the mice. A new thought is provided for treating and preventing chronic cold stress of livestock in the cold region and improving health and welfare of the livestock, and technical support is provided for improving the product quality of livestock products, reducing antibiotic dependence, widening the utilization channel of local agricultural and sideline products and developing functional feed additives for the cold region.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the application of lycopene in the preparation of products that alleviate damage to the body caused by chronic cold exposure. Background Technology

[0002] Chronic cold exposure leads to cold stress (CS). The pathogenesis of cold stress is complex, involving almost all tissues and organs throughout the body. Based on the intensity and duration of cold stress, it can be divided into acute cold stress (10-15°C lower than normal temperature, lasting from several minutes to a day) and chronic cold stress (lasting from a day to a week). Chronic cold exposure forces the animal's body into a high-metabolic, high-energy-consumption state to combat the cold environment and maintain body temperature and homeostasis. During chronic cold exposure, the animal's body often undergoes adaptive responses to resist the cold environment, including changes in blood pressure, appetite, and food intake. Under chronic cold exposure, the animal's endocrine system also enhances metabolic heat production by upregulating the levels of catecholamines and thyroid hormones.

[0003] Chronic cold exposure reduces animal productivity and shortens their reproductive cycle, severely impacting the livestock industry. During chronic cold exposure, meat-producing animals like pigs primarily use feed for warmth, leading to decreased daily weight gain, prolonged time to market, and significantly reduced meat quality. Chronic cold exposure also affects reproductive performance. In male animals, sperm motility and production are affected, resulting in decreased semen quality. In female animals, it disrupts the hypothalamic-pituitary-gonadal axis, affecting the estrous cycle. Furthermore, the survival rate of young animals is drastically reduced. As a persistent stressor, chronic cold exposure damages the animal's immune function, inducing oxidative stress, suppressing immune function, and harming gut health. The impact of chronic cold exposure on livestock production is substantial; therefore, finding an inexpensive, safe, and readily available drug to combat chronic cold exposure is essential for the livestock industry.

[0004] Lycopene (LYC) is a carotenoid widely found in plants and is one of the most potent natural antioxidants. Lycopene is inexpensive and readily available; it can be extracted from tomato scraps, animal feed residues, and substandard tomato fruit, thus possessing strong potential for widespread application. Previous studies have shown that lycopene has a positive effect on infectious diseases, such as those caused by Staphylococcus aureus and influenza viruses. However, there are no reports on its ability to alleviate damage caused by chronic cold exposure. Summary of the Invention

[0005] To address the aforementioned problems, this invention provides the application of lycopene in the preparation of products that alleviate damage caused by chronic cold exposure. This invention discovers that lycopene can alleviate inflammatory responses, reduce oxidative stress levels, and enhance the immune function of mice chronically exposed to cold.

[0006] To achieve the above objectives, the present invention provides the following technical solution: This invention provides the application of lycopene in the preparation of products that alleviate damage to the body caused by chronic cold exposure.

[0007] Preferably, the relief of damage caused by chronic cold exposure includes relieving one or more of the following: inflammatory response, oxidative stress, and immune function impairment caused by chronic cold exposure.

[0008] Preferably, alleviating the inflammatory response caused by chronic cold exposure includes reducing the levels of pro-inflammatory factors in the body.

[0009] Preferably, the pro-inflammatory factors include tumor necrosis factor α and / or interleukin 1β.

[0010] Preferably, alleviating oxidative stress caused by chronic cold exposure includes one or more of the following: increasing the body's total antioxidant capacity, the content of superoxide dismutase, and reducing the content of malondialdehyde in the body.

[0011] Preferred methods for alleviating immune function damage caused by chronic cold exposure include: increasing the body's immunoglobulin levels.

[0012] Preferably, the immunoglobulins include one or more of immunoglobulin A, immunoglobulin G, and immunoglobulin M.

[0013] Preferably, the product includes a drug or a feed additive.

[0014] Preferably, the unit dose of lycopene in the product is 5-15 mg / kg based on the weight of mice.

[0015] Preferably, the unit dose of lycopene in the product is 5-10 mg / kg based on the weight of mice.

[0016] Beneficial effects: This invention provides the application of lycopene in the preparation of products that alleviate damage caused by chronic cold exposure. This invention discovers that lycopene can alleviate inflammatory responses, reduce oxidative stress levels, and improve immune function in mice chronically cold-exposed. This provides new ideas for the treatment and prevention of chronic cold stress in livestock in cold regions and for improving livestock health and welfare. It also provides technical support for improving the quality of livestock products, reducing antibiotic dependence, expanding the utilization channels of local agricultural products, and developing functional feed additives for cold regions. Detailed Implementation

[0017] This invention provides the application of lycopene in the preparation of products that alleviate damage to the body caused by chronic cold exposure.

[0018] As one implementation method, the mitigation of damage to the body caused by chronic cold exposure includes mitigating one or more of the following: inflammatory response, oxidative stress, and immune function impairment caused by chronic cold exposure.

[0019] As one approach, alleviating the inflammatory response caused by chronic cold exposure includes reducing the levels of pro-inflammatory factors in the body.

[0020] In one implementation, the pro-inflammatory factors include tumor necrosis factor α and / or interleukin 1β.

[0021] As one implementation method, alleviating oxidative stress in the body caused by chronic cold exposure includes one or more of the following: increasing the body's total antioxidant capacity, the content of superoxide dismutase, and reducing the content of malondialdehyde in the body.

[0022] As one approach, alleviating immune function damage caused by chronic cold exposure includes increasing the body's immunoglobulin levels.

[0023] In one embodiment, the immunoglobulins include one or more of immunoglobulin A, immunoglobulin G, and immunoglobulin M.

[0024] In one implementation, the product includes a drug or feed additive.

[0025] In one embodiment, the unit dose of lycopene in the product is 5-15 mg / kg based on the body weight of mice. In another embodiment, the unit dose of lycopene in the product is 5-10 mg / kg based on the body weight of mice.

[0026] This invention investigates the adaptive changes in mice under chronic cold stress by detecting changes in food intake, water consumption, and body weight. It also uses ELISA to detect the levels of serum inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in mice after chronic cold exposure and after lycopene intervention, exploring the effects of chronic cold exposure on the inflammatory response in mice and the therapeutic effect of lycopene on the inflammatory response induced by chronic cold exposure. Furthermore, it uses ELISA to detect serum antioxidant-related indicators such as total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice after chronic cold exposure and after lycopene intervention. This study investigated the effects of chronic cold exposure on oxidative stress in mice and the therapeutic effect of lycopene on oxidative stress induced by chronic cold exposure in mice. Furthermore, the study used ELISA to detect the levels of serum immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in mice after chronic cold exposure and after lycopene intervention, exploring the effects of chronic cold exposure and lycopene intervention on mouse immunity.

[0027] Experimental results showed that chronic cold exposure induced adaptive changes in mice, leading to increased food intake and body weight. Furthermore, chronic cold exposure enhanced the inflammatory response in mice by increasing the levels of pro-inflammatory factors TNF-α and IL-1β in their serum, while lycopene intervention alleviated the inflammatory response by decreasing the levels of TNF-α and IL-1β in their serum. Chronic cold exposure also induced oxidative stress in mice by decreasing the levels of antioxidant markers T-AOC and SOD in their serum and increasing the levels of MDA in their serum. Lycopene intervention, by increasing the levels of T-AOC and SOD in their serum and decreasing the levels of MDA in their serum, treated the oxidative stress induced by chronic cold exposure in mice. In addition, this invention also found that chronic cold exposure affects the immune system of mice by decreasing the levels of immune markers IgA, IgG, and IgM in their serum, thus reducing their immunity. Lycopene intervention, by increasing the levels of IgA, IgG, and IgM in their serum, counteracted and treated the damage to the immune system caused by chronic cold exposure, thereby enhancing the immunity of mice. This invention provides new ideas for treating and preventing chronic cold stress in livestock in cold regions and improving livestock health and welfare. It also provides technical support for improving the quality of livestock products, reducing antibiotic dependence, expanding the utilization channels of local agricultural and sideline products, and developing functional feed additives for cold regions.

[0028] To further illustrate the present invention, the following detailed description, in conjunction with embodiments, illustrates the application of lycopene provided by the present invention in the preparation of products that alleviate damage to the body caused by chronic cold exposure. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0029] Example 1 Twenty mice were randomly divided into groups of 10 each, as follows: Control group (normal room temperature group): C57 / BL male mice (purchased from Charles River Lab), weighing 22-24g, 5 weeks old, were fed normal feed daily and had free access to sterile water. They were housed for 1, 2 and 3 weeks under controlled environmental conditions (temperature 24℃±2℃, humidity 40% and 12h light / dark cycle).

[0030] Chronic cold exposure group: C57 / BL male mice (purchased from Charles River Lab), weighing 22-24g, 5 weeks old, were fed normal feed daily, with free access to sterile water, and kept under controlled environmental conditions (40% humidity, 12h light / dark cycle). After 7 days, they were transferred to a 4℃ artificial climate chamber, maintained for 4h daily, and then returned to room temperature for 1, 2, and 3 weeks.

[0031] The food intake, water consumption, and body weight of the two groups of mice were measured and analyzed at different feeding times. The measurement methods are as follows: Feed and water normally at 8:00 AM daily, recording the total amount of feed (M1) and water (V1) given in the morning. Record the remaining feed (M2) and water consumption (V2) each evening. Calculate feed intake and water consumption using the following formula: Food intake = M1 - M2; Water intake = V1 - V2. The results are shown in Table 1-3.

[0032] Table 1. Food intake (g) of mice in different groups at different feeding times.

[0033] Note: In the table The difference between groups was significant. P <0.05), The difference between groups was highly significant. P <0.01) The difference between treatment groups was not statistically significant if the letters following the numerical values ​​were the same or there was no suffix after the numerical values. P (>0.05), the results for 1 week, 2 weeks and 3 weeks are the daily average ± standard error of feeding for 1 week, 2 weeks and 3 weeks, respectively, the same in the table below.

[0034] Table 2. Water and food intake (mL) of mice in different groups at different feeding times.

[0035] Table 3. Body weight (g) of mice in different groups at different feeding times.

[0036] As shown in Table 1, compared with the mice in the normal room temperature group, the food intake of the mice in the chronic cold exposure group increased significantly in the first and second weeks. P<0.05 The increase was extremely significant in the third week. P<0.01 This indicates that mice exposed to chronic cold increase their food intake to improve their body weight, thereby mitigating the effects of adverse environmental factors. Increased food intake is also an adaptive response undertaken by mice as homeothermic animals.

[0037] As shown in Table 2, compared with the mice in the normal room temperature group, the mice in the chronic cold exposure group did not have a significant change in water intake. This indicates that chronic cold exposure did not change the water metabolism balance of the mice, but maintained the body fluid homeostasis through other pathways.

[0038] As shown in Table 3, compared with the mice in the normal room temperature group, the body weight of the mice in the chronic cold exposure group was significantly increased in the second and third weeks after stimulation. P<0.05This indicates that mice exposed to chronic cold exhibited a compensatory response. In order to maintain homeostasis and cope with chronic cold exposure, the mice increased their body weight, which, through fat accumulation, served to keep them warm and protect them, thus achieving a compensatory balance.

[0039] Example 2 Forty male C57 / BL mice were randomly divided into groups of 10 each, as follows: The control group mice were fed normally without any treatment; Mice in the chronic cold exposure group were placed in a 4°C artificial climate chamber for 4 hours of low-temperature stimulation daily, and were fed normally. Chronic cold exposure + LYC1 group was placed in a 4℃ artificial climate chamber for 4 hours of low-temperature stimulation daily, fed normally, and gavaged with 5mg / kg / d of lycopene. The chronic cold exposure group + LYC2 was placed in a 4℃ artificial climate chamber for 4 hours of low-temperature stimulation daily, fed normally, and gavaged with 10mg / kg / d of lycopene.

[0040] Seven days later, blood was collected from the fundus venous plexus of mice. The blood was left to stand at room temperature for 30 minutes, then centrifuged at 3000 r / min for 5 minutes at 4°C. The supernatant was collected into a sterile ep tube, which is the mouse serum.

[0041] The levels of inflammatory factors TNF-α, IL-1β, and IL-6 in the collected mouse serum were detected using an ELISA kit (purchased from Thermo Fisher Scientific). The steps are as follows: The 96-well plates pre-coated with TNF-α, IL-1β, and IL-6 were equilibrated at room temperature for 30 min, and then samples were added according to the kit instructions. After sample addition, the plates were covered with a membrane and incubated at 37°C for 90 min. The plates were then washed five times with 300 μL / well of PBS-0.05% Tween 20 and patted dry. Antibody detection was then performed by adding antibodies against TNF-α, IL-1β, and IL-6 and incubating at 37°C for 60 min, followed by washing as described above. Enzyme-labeled secondary antibody was then added, and the plates were incubated at 37°C in the dark for 30 min, followed by washing as described above. Finally, TMB substrate was added, and the reaction was allowed to proceed at room temperature in the dark for 15 min for color development. The reaction was terminated by adding 2M H₂SO₄ and gently shaking. The absorbance was read at 450 nm within 15 min after the reaction was terminated, and the data were analyzed. The results are shown in Table 4.

[0042] Table 4. Effects of lycopene on serum inflammatory markers in mice under chronic cold exposure conditions (pg / mL)

[0043] As shown in Table 4, compared with the control group, the serum TNF-α level in the chronic cold exposure group was significantly increased.P< 0.01 Lycopene intervention reduced TNF-α levels in the serum of chronically cold-exposed mice; compared with the control group, the serum IL-1β levels in the chronically cold-exposed group were significantly increased. P<0.01 The intervention of lycopene reduced the serum IL-1β level in mice with chronic cold exposure; compared with the control group, the serum IL-6 level in mice with chronic cold exposure did not change significantly. This indicates that chronic cold exposure can induce an inflammatory response in the body, while the intervention of lycopene reversed the inflammation induced by chronic cold exposure, suggesting that lycopene has an anti-inflammatory effect and a therapeutic effect on chronic cold exposure in mice.

[0044] Example 3 The grouping and processing methods are the same as in Example 2.

[0045] Seven days later, blood was collected from the fundus venous plexus of mice. The blood was left to stand at room temperature for 30 minutes, then centrifuged at 3000 r / min for 5 minutes at 4°C. The supernatant was collected into a sterile ep tube, which is the mouse serum.

[0046] The total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and malondialdehyde (MDA) levels in the collected mouse serum were measured using an ELISA kit (purchased from Thermo Fisher Scientific). The steps are as follows: The 96-well plates pre-coated with T-AOC, SOD, and MDA were equilibrated at room temperature for 30 min, and then samples were added according to the kit instructions. After sample addition, the plates were covered with a membrane and incubated at 37°C for 90 min. The plates were then washed five times with 300 μL / well of PBS-0.05% Tween 20 and patted dry. Antibody detection was then performed by adding antibodies against T-AOC, SOD, and MDA and incubating at 37°C for 60 min, followed by washing as described above. Enzyme-labeled secondary antibody was then added, and the plates were incubated at 37°C in the dark for 30 min, followed by washing as described above. Finally, TMB substrate was added, and the reaction was allowed to proceed at room temperature in the dark for 15 min for color development. The reaction was terminated by adding 2M H₂SO₄ and gently shaking. The absorbance was read at 450 nm within 15 min after the reaction was terminated, and the data were analyzed. The results are shown in Table 5.

[0047] Table 5. Effects of lycopene on serum antioxidant indices in mice under chronic cold exposure conditions.

[0048] As shown in Table 5, compared with the control group, the T-AOC of mice in the chronic cold exposure group was significantly reduced ( P<0.01 Lycopene intervention reversed the effect of chronic cold exposure on T-AOC in mice; compared with the control group, the SOD content of mice in the chronic cold exposure group was significantly reduced (P<0.01 Lycopene intervention increased serum SOD levels in chronically cold-exposed mice; compared with the control group, serum MDA levels in the chronically cold-exposed group were significantly increased. P<0.01 Lycopene intervention reduced serum MDA levels in chronically cold-exposed mice. This indicates that chronic cold exposure can lead to a decrease in the antioxidant capacity of mice and promote oxidative stress. Lycopene, on the other hand, can alleviate and treat oxidative stress caused by chronic cold exposure by increasing the levels of T-AOC and SOD in mouse serum and decreasing the level of MDA, thereby protecting the bodies of chronically cold-exposed mice.

[0049] Example 4 The grouping and processing methods are the same as in Example 2.

[0050] Seven days later, blood was collected from the fundus venous plexus of mice. The blood was left to stand at room temperature for 30 minutes, then centrifuged at 3000 r / min for 5 minutes at 4°C. The supernatant was collected into a sterile ep tube, which is the mouse serum.

[0051] The levels of serum antibodies IgA, IgM, and IgG in the collected mouse serum were detected using an ELISA kit (purchased from Thermo Fisher Scientific). The steps are as follows: The 96-well plates pre-coated with IgA, IgM, and IgG were equilibrated at room temperature for 30 min, and then samples were added according to the kit instructions. After sample addition, the plates were covered with a membrane and incubated at 37°C for 90 min. The plates were then washed five times with 300 μL / well of PBS-0.05% Tween 20 and patted dry. Antibody detection was then performed by adding IgA, IgM, and IgG antibodies and incubating at 37°C for 60 min, followed by washing as described above. Enzyme-labeled secondary antibody was then added, and the plates were incubated at 37°C in the dark for 30 min, followed by washing as described above. Finally, TMB substrate was added, and the reaction was allowed to proceed at room temperature in the dark for 15 min for color development. The reaction was terminated by adding 2M H₂SO₄ and gently shaking. The absorbance was read at 450 nm within 15 min after the reaction was terminated, and the data were analyzed. The results are shown in Table 6.

[0052] Table 6. Effects of lycopene on serum immune parameters in mice under chronic cold exposure conditions (g / L)

[0053] As shown in Table 6, compared with the control group, the serum IgA content of mice in the chronic cold exposure group was significantly reduced ( P<0.01 Lycopene intervention increased the serum IgA content in chronically cold-exposed mice; compared with the control group, the serum IgG content in the chronically cold-exposed group was significantly reduced. P<0.01Lycopene intervention increased the serum IgG content in chronically cold-exposed mice; compared with the control group, the serum IgM content in the chronically cold-exposed group was significantly reduced. P<0.01 The intervention with lycopene increased the level of IgM in the serum of mice with chronic cold exposure. This indicates that chronic cold exposure can affect the immune function of mice and reduce their immunity, while lycopene can improve the immune function of mice by increasing the levels of IgA, IgG, and IgM antibodies in the serum, thereby playing a role in treating CCS in mice.

[0054] In conclusion, lycopene can alleviate the inflammatory response, reduce oxidative stress levels, and improve the immune function of mice chronically exposed to cold.

[0055] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of lycopene in the preparation of products that alleviate damage to the body caused by chronic cold exposure.

2. The application according to claim 1, characterized in that, The relief of damage caused by chronic cold exposure includes alleviating one or more of the following: inflammatory response, oxidative stress, and immune function impairment caused by chronic cold exposure.

3. The application according to claim 2, characterized in that, Alleviating the inflammatory response caused by chronic cold exposure includes reducing the levels of pro-inflammatory factors in the body.

4. The application according to claim 3, characterized in that, The pro-inflammatory factors include tumor necrosis factor α and / or interleukin 1β.

5. The application according to claim 2, characterized in that, Relieving oxidative stress caused by chronic cold exposure includes one or more of the following: increasing the body's total antioxidant capacity, superoxide dismutase levels, and reducing malondialdehyde levels.

6. The application according to claim 2, characterized in that, Relieving immune function damage caused by chronic cold exposure includes increasing the body's immunoglobulin levels.

7. The application according to claim 6, characterized in that, The immunoglobulins include one or more of immunoglobulin A, immunoglobulin G, and immunoglobulin M.

8. The application according to claim 1, characterized in that, The products include pharmaceuticals or feed additives.

9. The application according to claim 1 or 8, characterized in that, Based on the body weight of mice, the unit dose of lycopene in the product is 5~15mg / kg.

10. The application according to claim 9, characterized in that, Based on the body weight of mice, the unit dose of lycopene in the product is 5~10 mg / kg.

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