Composition and application thereof as well as food or medicine containing composition for delaying skeletal muscle senescence

Abstract

The invention provides a composition and application thereof, and a food or a medicine containing the composition and used for delaying skeletal muscle senescence. The composition comprises lactoferrin and pyrroloquinoline quinone disodium salt in a weight ratio of (20: 1)-(30: 1). The composition can be used for preparing foods or medicines for delaying skeletal muscle senescence, has the application of preventing and delaying sarcopenia, and has a good application prospect for preventing sarcopenia.

Description

Technical Field

[0001] The present invention relates to the technical field of functional compositions, and in particular to a composition and its application, and a food or medicine comprising the composition for delaying skeletal muscle aging. Background Art

[0002] Sarcopenia is a geriatric syndrome characterized by a progressive loss of muscle mass, strength, and function. In clinical practice, it is widely believed that sarcopenia is closely associated with an increased risk of falls, disability, and death. Due to muscle decline, the risk of falls and injuries increases with age, impacting not only physical health but also mental and social well-being. Therefore, delaying skeletal muscle aging and thereby preventing and treating sarcopenia has become a pressing issue for researchers.

[0003] Lactoferrin is widely present in mammalian milk and is the core immune protein in breast milk. It has multiple biological activities, including antiviral infection, antioxidant, antibacterial, anticancer, anti-inflammatory and immune regulation.

[0004] At present, there are no reports on the effects of lactoferrin and pyrroloquinoline quinone disodium salt (PQQ) on skeletal muscle aging, so further research and exploration of the specific relationship between them is of great significance. Summary of the Invention

[0005] To solve the above technical problems, the purpose of the present invention is to provide a composition and its application, as well as a food or medicine containing the composition for delaying skeletal muscle aging. The lactoferrin and pyrroloquinoline quinone disodium salt composition can improve muscle strength, muscle mass, muscle function, etc., and the combined intervention has a synergistic technical effect, thereby effectively delaying skeletal muscle aging.

[0006] To achieve the above object, the present invention first provides a composition, wherein the composition comprises lactoferrin and pyrroloquinoline quinone disodium salt, and the weight ratio of lactoferrin to pyrroloquinoline quinone disodium salt is 20:1-30:1.

[0007] In the above composition, preferably, the weight ratio of lactoferrin to pyrroloquinoline quinone disodium salt is 22:1-28:1, more preferably 25:1.

[0008] In the above composition, preferably, the source of the lactoferrin is one or a combination of two or more of cow's milk, goat's milk, cow's milk products and goat's milk products. More preferably, the source of the lactoferrin is one or a combination of two or more of lactoferrin powder, whole milk powder, skim milk powder and whey protein powder.

[0009] In the above composition, preferably, the purity of the pyrroloquinoline quinone disodium salt (PQQ) (on a dry basis) is ≥98.0 g / 100 g, that is, it should comply with the requirements of the New Food Ingredient Announcement (No. 8 of 2023), and the content of pyrroloquinoline quinone disodium salt (on a dry basis) is ≥98.0 g / 100 g.

[0010] The present invention also provides the use of the above composition in preparing a food capable of improving muscle attenuation and / or promoting myogenic differentiation. Preferably, the improvement of muscle attenuation is achieved by improving muscle quality, muscle strength and / or muscle function.

[0011] The present invention also provides a food for delaying skeletal muscle aging, which contains the above composition.

[0012] According to a specific embodiment of the present invention, preferably, the food is a health food.

[0013] The present invention also provides a medicine for delaying skeletal muscle aging, which contains the above composition.

[0014] The present invention also provides the use of the above-mentioned food or medicine in improving muscle quality, muscle strength and muscle function.

[0015] The technical solution of the present invention has the following beneficial technical effects:

[0016] The present invention provides a new use of a specific ratio formula of lactoferrin and pyrroloquinoline quinone disodium salt combination intervention in delaying sarcopenia, providing a new method for preventing sarcopenia. The new method is more effective than the existing technology within a specific ratio range and has good application prospects for the prevention of sarcopenia. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the grouping and drug administration of animal experiments.

[0018] Figure 2 Figure 3 is the body weight change curve of mice in each group during the intervention process.

[0019] Figure 3 The data of lean meat content of each group of mice are shown in Figure 2.

[0020] Figure 4 The fat content data of each group of mice are shown in Figure 2.

[0021] Figure 5 The skeletal muscle mass and index data results after sampling from each group of mice.

[0022] Figure 6 These are the grip strength test data results of mice in each group.

[0023] Figure 7These are the data results of the inverted grid hanging experiment for each group of mice.

[0024] Figure 8 These are the treadmill test data results for each group of mice. DETAILED DESCRIPTION

[0025] In order to have a clearer understanding of the technical features, objectives and beneficial effects of the present invention, the technical solution of the present invention is now described in detail below, but it should not be understood as limiting the scope of implementation of the present invention.

[0026] The present invention provides a composition composed of lactoferrin and pyrroloquinoline quinone disodium salt. The composition improves muscle strength, muscle mass, and muscle function in mice, and delays skeletal muscle aging. The efficacy experiments are as follows:

[0027] 1) Animal experiments

[0028] Six-week-old, SPF-grade male C57BL / 6 mice, weighing 20 ± 1 g each, were purchased from Speicherstadt. They were housed at 22 ± 2°C with a 12-h light-dark cycle and allowed to acclimate for one week.

[0029] Preparation of experimental drugs:

[0030] aD-Galactose: Precision weigh D-galactose powder (Sigma, A3134) and inject it intraperitoneally at a dose of 500 mg / kg / day based on the mouse's body weight. Dissolve it in distilled water. The intraperitoneal injection volume per mouse is 100 μL, and the solvent concentration is 100 mg / mL.

[0031] b. Lactoferrin: Accurately weigh lactoferrin powder and administer it orally to mice at a dose of 500 mg / kg / day based on their body weight. Slowly dissolve the powder in distilled water in small portions and magnetically stir at 55°C for 5 minutes to allow for complete dissolution. Each mouse should receive a volume of 100 μL of the solution at a solvent concentration of 100 mg / mL.

[0032] c. Pyrroloquinoline quinone disodium salt (PQQ): Accurately weigh pyrroloquinoline quinone disodium salt (PQQ) powder (CAS 122628-50-6) and administer to mice orally at a dose of 20 mg / kg / day based on body weight. Dissolve the powder in distilled water until fully dissolved. The gavage volume per mouse is 100 μL, and the solvent concentration is 4 mg / mL.

[0033] d. Lactoferrin and Pyrroloquinoline Quinone Disodium Salt (PQQ) Mixed Solution: Accurately weigh a certain amount of lactoferrin and Pyrroloquinoline Quinone Disodium Salt (PQQ) powder (CAS 122628-50-6) at a weight ratio of 25:1. Dissolve the mixture thoroughly using the above-described dissolution method. The oral volume for each mouse was 100 μL.

[0034] Animal Grouping:

[0035] Six-week-old male SPF C57BL / 6 mice were randomly divided into five groups, with six mice in each group, namely normal control group, sarcopenia model group, lactoferrin group, pyrroloquinoline quinone disodium salt group (PQQ group), and lactoferrin and pyrroloquinoline quinone disodium salt combined intervention group (combined group). The normal control group received daily intraperitoneal injection and oral gavage of normal saline, the other four groups received daily intraperitoneal injection of D-galactose, the sarcopenia model group received daily oral gavage of normal saline, the lactoferrin group received daily oral gavage of lactoferrin solution, the PQQ group received daily oral gavage of pyrroloquinoline quinone disodium salt solution, and the combined group received daily oral gavage of a mixed solution of lactoferrin and pyrroloquinoline quinone disodium salt for 8 weeks. Each group of mice received intraperitoneal and oral administration once a day, and the experimental period was 56 days. During the experiment, mice in each group had free access to food and water, and the mice were weighed regularly every week. The animal experimental grouping and drug administration are shown in the table. Figure 1 .

[0036] Test materials:

[0037] After the last oral administration of the drug to the mice, behavioral experiments were performed immediately the next day. After the test, the skeletal muscles of the tibialis anterior (TA), soleus (SOL), extensor digitorum longus (EDL), and gastrocnemius (GAS) of the mouse hind limbs were obtained and weighed. The skeletal muscles of the left leg of the mouse 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.

[0038] 2) Body composition analysis

[0039] Body composition analysis and imaging

[0040] After the intervention, the mice's body composition was measured using the Awake Small Animal Body Composition Analysis and Imaging System (Animal Fat Meter). Before testing, the mice were weighed and placed in the instrument after a self-check. After the measurement, physiological parameters such as lean mass and fat content were obtained while the mice were awake. Each mouse was measured two to three times.

[0041] 3) Behavioral experiments

[0042] a. Inverted grid hanging experiment

[0043] The inverted net is a 43cm wire mesh consisting of 1mm diameter wire. It is surrounded by a 4cm deep wooden bearing (to prevent the occasional mouse from trying to climb to the other side). Each animal is placed in the center of the grid, then inverted 180° and placed 40cm 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 the mouse complete this test in an unknown state in a limited number of repetitions.

[0044] b. Grip strength test

[0045] The measurement was performed using a rat grip strength tester (model: ZS-ZL). The basic process is divided into the following steps:

[0046] (1) Install the elastic metal strip onto the sensor and secure it firmly.

[0047] (2) Turn on the sensor and select peak mode. Do not apply a force greater than the sensor's strength tolerance.

[0048] (3) The display sensor is reset to zero.

[0049] (4) Take the mouse out of the cage and grab the middle of the mouse's tail with your thumb and index finger.

[0050] (5) Limb measurement: The mouse was placed on an elastic metal bar, and before measurement, the mouse's front and hind paws were allowed to touch the elastic metal bar, with the torso and elastic metal bar kept horizontal. The mouse's tail was gently pulled from the top of the elastic metal bar until the mouse could no longer maintain its grip and the metal bar fell off its paws. During the test, the measuring device recorded the maximum limb muscle force value (g) and displayed it on the instrument screen. Each mouse was tested three times in a row.

[0051] c. Treadmill test

[0052] (1) Adaptation stage

[0053] Table 1

[0054] Speed (m / min) Acceleration time (s) Speed duration (min) Initial velocity 5 5 3 First speed 10 5 3 Secondary speed 15 5 4

[0055] Testing was performed using a small animal treadmill (model: ZS-PT-IV). The treadmill parameters for the acclimation phase are shown in Table 1. The treadmill stimulation current was set to 0.3 mA. This gradual training method allowed the animals to acclimate to the treadmill. This acclimation process lasted 2-3 days, once daily for approximately 10 minutes.

[0056] (2) Testing phase

[0057] Table 2

[0058] Speed (m / min) Acceleration time (s) Speed duration (min) Initial velocity 10 5 2 First speed 15 5 3 Secondary speed 20 5 to exhaustion

[0059] Treadmill parameters for the testing phase are shown in Table 2. The treadmill stimulation current was set to 0.3 mA. Exhaustion was indicated by the mice stopping running for 10 seconds and then failing to run after 5 seconds of electrical or noise stimulation. Time to exhaustion was recorded. This procedure was repeated daily for 2-3 days.

[0060] The present invention has completed animal level verification.

[0061] 4) Experimental results

[0062] 1. Figure 2 is the weight change curve of mice, Figure 2 It can be seen that the body weight of mice in each group showed an upward trend, and there was no significant difference between the groups.

[0063] 2. Figure 3 and Figure 4 The data are respectively the lean meat content and fat content of each group of mice. Figure 3 and Figure 4 Compared to the sarcopenia model group, mice treated with lactoferrin and PQQ, either alone or in combination, showed a significant increase in lean muscle mass, approaching that of the normal control group. This reduction in fat accumulation also slowed the accumulation of fat. This increase in lean muscle mass and decrease in fat mass indicate an increase in the proportion of muscle tissue, suggesting that the symptoms of muscle loss and / or atrophy have improved.

[0064] 3. Skeletal muscle index is the criterion for judging muscle atrophy. Figure 5 The skeletal muscle mass and index test results of each group of mice were collected. Figure 5It can be seen that the muscle index of the skeletal muscles of various parts of the hind limbs in the sarcopenia model group was significantly lower than that in the normal control group mice, indicating that the mice in the sarcopenia model group had aging-related muscle atrophy and the experimental model was successful. Compared with the normal control group mice, the skeletal muscle mass and index of the hind limb gastrocnemius muscle (GAS) (Figure A is the gastrocnemius muscle mass test result, Figure E is the gastrocnemius muscle index test result), tibialis anterior muscle (TA) (Figure B is the tibialis anterior muscle mass test result, Figure F is the tibialis anterior muscle index test result), extensor digitorum longus (EDL) (Figure C is the extensor digitorum longus muscle mass test result, Figure G is the extensor digitorum longus muscle index test result), and soleus muscle (SOL) (Figure D is the soleus muscle mass test result, Figure H is the soleus muscle index test result) of the sarcopenia model group mice were significantly reduced. After lactoferrin and PQQ intervention alone or in combination, the mass and index of the four parts of skeletal muscle all had different degrees of callback, and the corresponding skeletal muscle mass and index of the combined group were closer to or even higher than those of the normal control group, indicating that nutritional intervention has a certain improvement effect on the reduction of muscle mass, and the combined effect of lactoferrin and PQQ is more obvious. Compared with the intervention of lactoferrin or PQQ alone, the muscle weight and skeletal muscle index of the gastrocnemius (GAS), tibialis anterior (TA), extensor digitorum longus (EDL), and soleus (SOL) muscles of the hind limbs of mice in the combined group were significantly increased, suggesting that lactoferrin and PQQ have a synergistic effect in improving skeletal muscle mass and index.

[0065] 4. Figure 6 The following are the grip strength test results of each group of mice. Figure 6 As can be seen, compared to the sarcopenia model group, the maximum muscle strength values of mice in each group were significantly improved after treatment with lactoferrin and PQQ alone or in combination, and the combined group was significantly higher than the lactoferrin and PQQ groups, indicating that the combined intervention can better improve the muscle strength of sarcopenic mice. Compared with treatment with lactoferrin or PQQ alone, the maximum muscle strength values of mice in the combined group were significantly increased, suggesting that lactoferrin and PQQ have a synergistic effect in improving muscle strength.

[0066] 5. Figure 7 and Figure 8 The results of the inverted grid hanging test and the treadmill test for each group of mice are shown in Figure 2. Figure 7 、 Figure 8 As can be seen, compared to the sarcopenia model group, the time to exhaustion and the hanging time on the inverted grid in the treadmill test were prolonged in all groups of mice after either lactoferrin or PQQ intervention alone, with the combined group showing significantly longer times than the lactoferrin and PQQ groups, indicating that the combined intervention significantly improved the mice's muscle endurance and function. Compared to either lactoferrin or PQQ intervention alone, the combined group significantly increased both the time to exhaustion and the hanging time on the inverted grid, suggesting that lactoferrin and PQQ have a synergistic effect in improving muscle endurance and function.

[0067] Example 1

[0068] This embodiment provides a composition, wherein the composition can improve muscle attenuation and promote myogenic differentiation, and the composition includes lactoferrin and pyrroloquinoline quinone disodium salt, and the weight ratio of lactoferrin to pyrroloquinoline quinone disodium salt is 25:1.

[0069] Example 2

[0070] This embodiment provides a food for delaying skeletal muscle aging, which contains the composition provided in Example 1.

[0071] Example 3

[0072] This embodiment provides a drug for delaying skeletal muscle aging, which contains the composition provided in Example 1 and other necessary ingredients of the drug, such as excipients.

[0073] The contents not described in detail in this specification belong to the prior art known to those skilled in the art.

[0074] Finally, it should be noted that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art will be able to modify the technical solutions described in the aforementioned embodiments or substitute equivalents for some of the technical features. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A composition, wherein The composition comprises lactoferrin and pyrroloquinoline quinone disodium salt, wherein the weight ratio of the lactoferrin to the pyrroloquinoline quinone disodium salt is 20:1-30:

1.

2. The composition according to claim 1, wherein The weight ratio of lactoferrin to pyrroloquinoline quinone disodium salt is 22:1-28:

1.

3. The composition according to claim 1, wherein The source of the lactoferrin is one or a combination of two or more of cow's milk, goat's milk, cow's milk products and goat's milk products.

4. The composition according to claim 3, wherein The source of the lactoferrin is one or a combination of two or more of lactoferrin powder, whole milk powder, skim milk powder and whey protein powder.

5. The composition according to claim 1, wherein The purity of the pyrroloquinoline quinone disodium salt is ≥98.0 g / 100 g, calculated on a dry basis.

6. Use of the composition according to any one of claims 1 to 5 in the preparation of a food capable of improving muscle attenuation and / or promoting myogenic differentiation.

7. The use according to claim 6, wherein: The improvement of muscle attenuation is achieved by improving muscle mass, muscle strength and / or muscle function.

8. A food for delaying skeletal muscle aging, comprising the composition according to any one of claims 1 to 5.

9. The food for delaying skeletal muscle aging according to claim 8, wherein The food is a health food.

10. A medicine for delaying skeletal muscle aging, comprising the composition according to any one of claims 1 to 5.

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