Application of lychee fermentation product metabiotics in product for relieving muscle aging

Abstract

The invention specifically discloses an application of lychee fermentation product metabiotics in a product for relieving muscle aging. The lychee leavening plays a role as a common food raw material component, can shorten the period of relieving the sarcopenia symptom, and can significantly relieve the sarcopenia symptom.

Description

Technical Field

[0001] The invention relates to the field of food technology, and in particular to application of litchi fermentation postbiotics in products for alleviating muscle aging. Background Art

[0002] Sarcopenia refers to a syndrome in which muscle mass and strength gradually decrease with age, and the elderly population worldwide is affected. With the acceleration of the global aging process, the incidence of sarcopenia is on the rise. Sarcopenia not only affects the physical function and quality of life of the elderly, but may also bring a series of health problems and complications. For example, the reduction of muscle mass and strength can lead to slow movement, slower walking speed, and decreased balance ability, which significantly increases the risk of falls and leads to fractures. At the same time, sarcopenia often coexists with other geriatric syndromes (such as malnutrition, weakness, and cognitive dysfunction), forming a vicious circle and accelerating the health decline of the elderly. Therefore, the prevention and treatment of sarcopenia has become an important public health issue.

[0003] At present, muscle attenuation symptoms are generally relieved through exercise therapy, nutritional support therapy, muscle strengthening training and other methods, but the above methods have a long period of time and slow effect in relieving muscle attenuation. In recent years, searching for ingredients in plants that can relieve muscle attenuation symptoms has gradually become a research hotspot. my country has rich plant resources and a long history of use. There is a broad application prospect for extracting ingredients from plants that can relieve muscle attenuation symptoms. Summary of the invention

[0004] In order to shorten the period of alleviating muscle wasting symptoms and improve the effect of alleviating muscle wasting symptoms, the present invention provides an application of litchi fermentation postbiotics in a product for alleviating muscle aging.

[0005] In a first aspect, the present invention provides an application of litchi fermentation postbiotics in a product for alleviating muscle aging, using the following technical solution:

[0006] Application of a litchi fermentation product postbiotic in a product for alleviating muscle aging.

[0007] As a typical subtropical fruit, litchi has important economic significance. It is rich in flavonoids and phenolic acid substances. The Compendium of Materia Medica records that litchi has the effects of nourishing the liver and spleen, benefiting the liver and regulating qi, nourishing blood, warming the middle and relieving pain, and nourishing the heart and calming the mind. It can regulate qi, disperse knots, relieve pain, and prevent diarrhea. Pharmacological studies have shown that litchi kernel has the effects of lowering blood sugar, lowering blood lipids, anti-virus, and anti-liver damage. Traditional Chinese medicine theory believes that litchi kernel has the effects of promoting qi and dispersing knots, dispelling cold and relieving pain.

[0008] However, there are no reports on the effects of litchi fermentation on relieving skeletal muscle aging. On the one hand, based on the pharmacological effects disclosed in existing studies, litchi is widely used in lowering blood sugar, preventing liver damage and antiviral effects. Therefore, most of the existing studies on litchi focus on these related aspects. On the other hand, there are currently no plant-derived ingredients that are well targeted to effectively relieve the symptoms of muscle aging.

[0009] The present invention has found through a series of experimental studies that lychee fermented product can effectively and significantly alleviate the symptoms of muscle aging. Specifically, the lychee fermented product used in the present invention is used as a common food raw material ingredient to alleviate the symptoms of muscle aging, which helps to increase muscle weight and skeletal muscle index. It takes effect after 8 weeks of intervention at an appropriate dosage concentration, with a short intervention time and a significant effect in alleviating muscle attenuation. Based on this, the present invention proposes a new use of lychee fermented product in delaying muscle wasting disease, provides a new method for preventing muscle wasting disease, and provides good application prospects for clinically alleviating muscle wasting disease.

[0010] Preferably, the subject includes humans or animals.

[0011] Preferably, the present invention uses any one of D-gal model mice, naturally aging mice, and SAM rapid aging mice.

[0012] Preferably, the present invention uses D-gal model mice. D-gal (D-galactose) promotes the aging process of skeletal muscle by simulating some biochemical changes in the natural aging process, such as oxidative stress and cell apoptosis.

[0013] Preferably, alleviating muscle aging comprises increasing the muscle weight and / or skeletal muscle index of the gastrocnemius, extensor tibialis anterior, extensor digitorum longus, and soleus muscles.

[0014] Preferably, alleviating muscle aging includes improving at least one of liver index, kidney index, spleen index and lung health.

[0015] Preferably, alleviating muscle aging includes reducing at least one of the degree of atrophy, the number of vacuoles, and the diameter of vacuoles of muscle fibers.

[0016] The D-gal muscle aging model mice used in the present invention illustrate the improvement effect of litchi fermentation products on muscle attenuation after intervention from the perspectives of muscle strength, quality and endurance, liver coefficient, degree of muscle atrophy, etc., which has a great reference significance for the results of human clinical trials.

[0017] In a second aspect, the present invention provides a product for alleviating muscle aging, which adopts the following technical solution:

[0018] A product for alleviating muscle aging, comprising lychee fermentate.

[0019] Preferably, the dosage of the litchi fermented product is 1-40000 mg / kg / day.

[0020] Preferably, the dosage of the litchi fermented product is 26.1-130.5 mg / kg / day.

[0021] Preferably, the dosage of the litchi fermented product is at least one of 26.1 mg / kg / day (low dose), 78.3 mg / kg / day (medium dose), and 130.5 mg / kg / day (high dose).

[0022] Preferably, the dosage of the litchi fermented product is 78.3 mg / kg / day.

[0023] The present invention uses different doses of lychee fermented products to study the relief of muscle attenuation symptoms. The research results show that not all doses of lychee fermented products will improve muscle attenuation, nor does the higher the dose of lychee fermented products, the more significant the effect of relieving muscle attenuation. Instead, there is an optimal dosage range, and when the dose is 78.3 mg / kg / day, the relief effect on muscle attenuation symptoms is the best. At the same time, the present invention applies lychee fermented products to food and health food, and the effective dose is lower than the dosage of the composition for relieving muscle attenuation in the prior art. Therefore, the lychee fermented products in the present invention can effectively and significantly relieve muscle attenuation symptoms at a low effective dose.

[0024] Preferably, the product can be litchi fermentation product concentrate or litchi fermentation product powder (solid beverage), or a composition prepared with litchi fermentation product. The composition can be food or health food. The main dosage forms are oral liquid, beverage, solid beverage, and soft candy, which are mainly taken by direct drinking, mixing, or chewing.

[0025] By applying litchi fermentation products to products of different dosage forms, it is possible to provide products of suitable dosage forms for different groups and broaden the scope of application.

[0026] Preferably, the method for preparing the litchi fermentation product comprises the following steps:

[0027] S1, after mixing litchi juice, litchi pomace and sugar, inoculating yeast and / or lactic acid bacteria, performing a primary fermentation operation to obtain a primary fermentation filtrate;

[0028] S2, inoculating acetic acid bacteria into the primary fermentation filtrate, and performing a secondary fermentation operation to obtain the litchi fermentation product.

[0029] Preferably, the added amount of the sugar is 0%-5% of the total mass of the litchi juice and litchi pomace.

[0030] Preferably, in the primary fermentation operation, the sealed static fermentation time is 10-168h.

[0031] Preferably, in the secondary fermentation operation, the aeration fermentation time is 10-168h.

[0032] The secondary fermentation in the present invention can not only retain the effective ingredients in litchi for alleviating muscle attenuation symptoms to the greatest extent, but also improve the palatability of the prepared litchi fermented product, thus providing a basis for the application of the litchi fermented product in food. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 Effect of litchi fermentation on the intake of D-gal in aging mice Figure 1 A), weight ( Figure 1 B) impact.

[0034] Figure 2 Effect of litchi fermentation on liver function of D-gal aging mice Figure 2 A) Spleen Figure 2 B) Kidney ( Figure 2 C) impact.

[0035] Figure 3 To investigate the effects of litchi fermentation products on body composition of D-gal aging mice.

[0036] Figure 4 Effect of litchi fermentation on muscle weight of D-gal aging mice Figure 4 A- Figure 4 D)( Figure 4 E- Figure 4 H) and skeletal muscle index.

[0037] Figure 5 Effect of litchi fermentation on muscle strength of D-gal aging mice Figure 5 A) and endurance ( Figure 5 B. Figure 5 C) impact.

[0038] Figure 6 This is the effect diagram of mouse muscle fibers ( Figure 6 A is the Control group, Figure 6 B is the D-gal modeling group, Figure 6 C- Figure 6 E is litchi fermentation intervention group). DETAILED DESCRIPTION

[0039] For better understanding and implementation, the technical solution of the present invention will be clearly and completely described below in conjunction with embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0041] Unless otherwise indicated, all numerical values ​​expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as modified by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth herein are approximations that can vary depending upon the desired properties desired to be obtained.

[0042] As used herein, "and / or" means one or all of the mentioned elements.

[0043] As used herein, “including” and “comprising” encompasses the case where only the stated elements are present as well as the case where there are other elements other than the stated elements.

[0044] All percentages in the present invention are by weight, unless otherwise specified.

[0045] Unless otherwise indicated, "a", "an", "an" and "the" used in this specification are intended to include "at least one" or "one or more". For example, "a component" means one or more components, so more than one component may be considered and may be employed or used in the implementation of the embodiment.

[0046] 1. Preparation of litchi fermentation products

[0047] S1, washing and squeezing litchi fruits to obtain litchi juice and litchi pomace; mixing the litchi juice and litchi pomace with sugar (the sugar accounts for 3% of the total mass of the litchi juice and litchi pomace, and the sugar includes fructose and brown sugar in a mass ratio of 1:1), inoculating yeast (sake yeast, the addition amount of yeast is 0.9‰ of the total mass of the litchi juice and litchi pomace), sealing and standing for fermentation at 30°C for 72h, and obtaining a primary fermentation filtrate;

[0048] S2, inoculating acetic acid bacteria into the primary fermentation filtrate (acetic acid bacteria include Acetobacter aceticus and Acetobacter aceticus in a mass ratio of 1:1, and the amount of acetic acid bacteria added is 1‰ of the total mass of litchi juice and litchi pomace), and culturing at 15°C with ventilation (ventilation rate of 1 mL / min / mL) for 120 h to obtain litchi fermentation product.

[0049] 2. Animal experiment design

[0050] Six-week-old male SPF C57BL / 6 mice, weighing 23±1g / mouse, were purchased from Vitallife. The breeding conditions were room temperature 22±2℃, light and dark cycle 12h each, and they were adapted for one week after purchase.

[0051] Experimental drugs:

[0052] (1) D-galactose (D-gal): D-galactose powder (Sigma, A3134) was accurately weighed and dissolved in distilled water at a dose of 500 mg / kg / day according to the mouse body weight. The intraperitoneal injection volume of each mouse was 100 μL to induce the mouse aging model.

[0053] (2) Products containing the above-mentioned litchi fermented products: The mice were given an intervention dose of 78.3 mg / kg / day based on their body weight. The gavage volume for each mouse was 100 μL.

[0054] Animal Grouping:

[0055] Six-week-old male SPF-grade C57BL / 6 mice were randomly divided into three groups, with six mice in each group, namely a normal control group (control group), a D-gal aging group, and a product group containing litchi fermentation products.

[0056] The normal control group (control group) was intraperitoneally injected and gavaged with normal saline every day; the other two groups were intraperitoneally injected with D-galactose every day, the D-gal aging group was gavaged with normal saline every day, and the product group containing litchi fermentation products was gavaged with litchi postbiotic solution every day for 8 weeks. Each group of mice was intraperitoneally and gavaged once a day, and the experimental period was 56 days. During the experiment, the mice in each group were free to eat and drink water, and the mice were weighed regularly every week.

[0057] Test materials:

[0058] After the last oral administration of the drug to the mice, the behavioral experiment of mice was carried out immediately the next day. After the test, the skeletal muscles of the tibialis anterior (TA), soleus (SOL), extensor digitorum longus (EDL), gastrocnemius (GAS) and other hind limbs of the mice were taken and weighed. The skeletal muscles of the left leg of the mice were preserved in tissue fixative, and the skeletal muscles of the right leg were quickly frozen in liquid nitrogen and placed in a -80°C refrigerator for use.

[0059] 3. Body composition analysis and imaging

[0060] After the intervention, the body composition of mice was measured using the awake small animal body composition analysis and imaging system (animal fat meter). The mice were weighed before the test and then placed in the instrument after self-check. After the measurement, the physiological parameters such as lean meat content and fat content of the mice in the awake state were obtained. Each mouse was measured 2-3 times.

[0061] 4. Inverted grid suspension experiment

[0062] Each animal is placed in the center of the grid, then inverted 180° and placed 40 cm above the cage pad. A timer is used to record the time the animal is suspended on the wire. If the mouse falls within 10 seconds, the test should be repeated immediately to ensure accurate results. This test is based on the mouse's instinctive fear of falling, so it is best to let it complete this test in an unknown state in limited repetitions.

[0063] 5. Grip strength test

[0064] The mouse was placed on the elastic metal bar, and before the measurement, the mouse's front and hind paws were allowed to touch the elastic metal bar, so that the trunk and the elastic metal bar were kept horizontal, and the mouse's tail was gently pulled from the top of the elastic metal bar until the mouse could not maintain the grip and the metal bar fell off from its paws. During the test, the measuring device recorded the maximum limb muscle force value (g) and displayed it on the screen of the instrument. Each mouse was tested 3 times in a row.

[0065] 6. Treadmill test

[0066] (1) Adaptation phase: The treadmill parameter settings for the adaptation phase are shown in Table 1:

[0067] Table 1 Treadmill parameters during the adaptation phase

[0068] Speed(m / min) Acceleration time(s) Speed ​​duration (min) Initial Speed 5 5 3 First speed 10 5 3 Secondary speed 15 5 4

[0069] The stimulation current of the treadmill was set to 0.3 mA, and this gradual training method was used to adapt the animals to the treadmill. This adaptation process lasted 2-3 days, once a day, and each time was about 10 minutes.

[0070] (2) Test phase: The treadmill parameter settings in the test phase are shown in Table 2:

[0071] Table 2 Treadmill parameters during the test phase

[0072] Speed(m / min) Acceleration time(s) Speed ​​duration (min) Initial Speed 10 5 2 First speed 15 5 3 Secondary speed 20 5 To exhaustion

[0073] The treadmill stimulation current was set to 0.3 mA. The sign of exhaustion was that the mouse stopped running for 10 seconds, and then the animal was stimulated with electricity and noise for 5 seconds but still could not run. The exhaustion time of the mouse was recorded. This process was repeated once a day for 2-3 days.

[0074] 7. H&E staining of gastrocnemius muscle sections

[0075] The gastrocnemius muscle was fixed with 4% formaldehyde and then rinsed with PBS to prepare paraffin sections. The nuclei were stained with hematoxylin and the cytoplasm was stained with eosin. The sections were dehydrated with gradient ethanol and then sealed with neutral resin for observation under a microscope.

[0076] Example 1: Effects of litchi fermentation on body weight, food intake, liver, spleen and kidney of D-gal model aging mice

[0077] like Figure 1 As shown in A and 1B, there was no difference in food intake among the groups, and the body weight coefficient also showed an upward trend over time. Among them, the body weight coefficient of mice in the medium-dose litchi fermented product group increased most significantly, followed by the mice in the high-dose litchi fermented product group, while the body weight coefficient of mice in the low-dose litchi fermented product group was basically the same as that of the normal control group (Control group), indicating that medium and high-dose litchi fermented products are more conducive to the weight development of mice.

[0078] like Figure 2 A- Figure 2 As shown in Figure C, the liver coefficient, spleen coefficient and kidney coefficient of D-gal model mice decreased significantly compared with the control group. However, after intervention with litchi fermented products, the liver, spleen and kidney of each group of mice showed different degrees of recovery. Specifically, medium and high doses of litchi fermented products were more helpful in improving the liver coefficient of mice. Among them, under the intervention of medium dose of litchi fermented products, the liver coefficient was able to recover to the same level as the control group ( Figure 2 A); while lychee fermented food can also improve the spleen coefficient and kidney coefficient, but the difference in the dosage of lychee fermented food on the improvement of liver coefficient and kidney coefficient is not significant ( Figure 2 B. Figure 2 C), and the effect of medium-dose lychee fermentation on improving the liver coefficient was significantly higher than the effect of medium-dose lychee fermentation on improving the spleen coefficient and kidney coefficient. The above results indicate that lychee fermentation is safe and has a certain liver-protecting effect in the intervention process.

[0079] Example 2: Effects of litchi fermentation on body composition of D-gal model aging mice

[0080] like Figure 3 As shown in the figure, the lean meat content of D-gal model mice decreased compared with the control group, while the fat content increased compared with the control group. After intervention with litchi fermentation, the lean meat content of mice increased significantly compared with the D-gal model mice ( Figure 3 A), and the lean meat content of mice in the medium-dose litchi fermentation group returned to a level close to that of the control group; at the same time, the fat content of mice decreased significantly compared with the mice after D-gal modeling and was close to that of the control group ( Figure 3B), indicating that litchi fermentation intervention can delay fat accumulation in mice. The increase in lean meat content and the decrease in fat content in mice represent an increase in the proportion of muscle tissue in the mice, indicating that the development and growth of skeletal muscle in mice have improved; indicating that medium-dose litchi fermentation can take effect after continuous use for 8 weeks (56 days), which can greatly shorten the period of relieving muscle wasting symptoms and can alleviate the symptoms of muscle wasting to a certain extent.

[0081] Example 3: Litchi fermented product alleviates muscle mass loss in D-gal model aging mice

[0082] Figure 4 The skeletal muscle plasmids and indexes of each part of the mice in each group are shown in Table 1. The skeletal muscle index is the criterion for judging muscle atrophy. Figure 4 As shown, it can be seen that the muscle weight and skeletal muscle index of the gastrocnemius (GAS), extensor tibialis anterior (TA), extensor digitorum longus (EDL), and soleus (SOL) of the D-gal modeled mice were significantly reduced compared with those of the Control group mice, indicating that aging-related muscle atrophy occurred in the D-gal group mice and the experimental model was successful.

[0083] After intervention with litchi fermented products, the mass and index of the above four parts of skeletal muscle recovered to varying degrees. Specifically, the mass and index of gastrocnemius, tibialis anterior and extensor digitorum longus recovered most significantly under the action of medium dose of litchi fermented products, while the mass and index of soleus recovered most significantly under the action of high dose of litchi fermented products ( Figure 4 D. Figure 4 H), among which, the mass and index of the tibialis anterior extensor muscle of mice in the intervention group with medium-dose litchi fermentation product were higher than those in the control group ( Figure 4 B. Figure 4 F), indicating that medium-dose lychee fermented products can be effective after continuous use for 8 weeks (56 days), which can greatly shorten the period of relieving muscle wasting symptoms. Nutritional intervention has a certain improvement effect on the reduction of muscle mass.

[0084] Example 4: Litchi fermented product improves muscle strength and endurance in D-gal model aging mice

[0085] Figure 5 A is the result of the gripping strength experiment; the gripping strength, grid inversion time and treadmill exhaustion time of D-gal model mice were significantly reduced compared with the control group. After intervention with litchi fermented products, the gripping strength of mice was significantly improved, and the medium-dose litchi fermented products had the most significant effect on improving the gripping strength of mice.

[0086] Figure 5 B is the results of the inverted grid hanging experiment of each group of mice. Figure 5C is the result of the treadmill experiment of each group of mice; the grid inversion time and treadmill exhaustion time of the D-gal model mice were significantly reduced compared with the Control group. After intervention with litchi fermented products, the grid inversion time and treadmill exhaustion time of the mice were prolonged. Specifically, the grid inversion time of the mice intervened with medium and high doses of litchi fermented products was able to return to the same level as the Control group mice, and even the treadmill exhaustion time of the mice intervened with medium doses of litchi fermented products was able to exceed the treadmill exhaustion time of the Control group mice, which shows that the intervention with medium doses of litchi fermented products can alleviate the endurance decline caused by muscle attenuation in mice to the greatest extent.

[0087] In summary, medium-dose litchi fermented products can take effect after 8 weeks (56 days) of continuous use, which can not only greatly shorten the period of relieving muscle wasting symptoms, but also improve the muscle strength of mice with muscle wasting and greatly improve the muscle endurance of mice.

[0088] Example 5: Litchi fermented product improves muscle fiber in D-gal model aging mice

[0089] Figure 6 This is the gastrocnemius muscle stained with H&E; it can be seen that D-gal model mice ( Figure 6 B) compared to the control group ( Figure 6 A) The muscle fibers of mice showed obvious atrophy. After intervention with litchi fermented products ( Figure 6 C- Figure 6 E), the muscle fiber atrophy of mice was significantly improved, indicating that litchi fermented products can significantly improve the symptoms of muscle attenuation.

[0090] The above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention is described in detail with reference to the above embodiments, ordinary technicians in the relevant field should understand that the technical solutions of the present invention can be modified or equivalently replaced, but these modifications or replacements are all within the protection scope of the present invention.

Claims

1. Application of a litchi fermentation product postbiotic in a product for alleviating muscle aging.

2. The use of litchi fermentation postbiotics in products for alleviating muscle aging as claimed in claim 1, characterized in that: The subjects include humans or animals.

3. The use of litchi fermentation postbiotics in products for alleviating muscle aging as claimed in claim 1, characterized in that: Alleviating muscle aging includes increasing the muscle weight and / or skeletal muscle index of the gastrocnemius, extensor tibialis anterior, extensor digitorum longus, and soleus muscles.

4. The use of litchi fermentation postbiotics in products for alleviating muscle aging as claimed in claim 1, characterized in that: Alleviating muscle aging includes improving at least one of liver coefficient, kidney coefficient, spleen coefficient and lung health.

5. The use of litchi fermentation postbiotics in products for alleviating muscle aging as claimed in claim 1, characterized in that: Alleviating muscle aging includes reducing at least one of the degree of atrophy, the number of vacuoles, and the diameter of vacuoles of muscle fibers.

6. A product for alleviating muscle aging, characterized in that: Includes lychee ferment.

7. The product for alleviating muscle aging according to claim 6, characterized in that: The dosage of the litchi fermented product is 1-40000 mg / kg / day.

8. The product for alleviating muscle aging according to claim 7, characterized in that: The dosage of the litchi fermented product is 26.1-130.5 mg / kg / day.

9. The product for alleviating muscle aging according to claim 6, characterized in that: The product includes any one of food, pet food, health food and daily chemical products.

10. The product for alleviating muscle aging according to claim 6, characterized in that: The preparation method of the litchi fermented product comprises the following steps: S1, after mixing litchi juice, litchi pomace and sugar, inoculating yeast and / or lactic acid bacteria, performing a primary fermentation operation to obtain a primary fermentation filtrate; S2, inoculating acetic acid bacteria into the primary fermentation filtrate, and performing a secondary fermentation operation to obtain the litchi fermentation product.

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