Application of ginseng oral liquid in improving muscle function decline caused by aging

Abstract

This invention belongs to the field of traditional Chinese medicine technology, specifically relating to the application of Ginseng Strengthening Oral Liquid in improving age-related muscle function decline. This invention, through research, is the first to discover that Ginseng Strengthening Oral Liquid can effectively improve age-related muscle strength decline, reduce collagen deposition, significantly improve muscle quality, and improve skeletal muscle damage, thereby effectively addressing age-related skeletal muscle function decline. It can be used to develop effective prevention and intervention strategies for age-related skeletal muscle function and quality decline. Furthermore, while addressing age-related muscle function decline, Ginseng Strengthening Oral Liquid does not cause kidney damage, metabolic disorders, or abnormal liver and kidney weight, demonstrating good systemic safety. Therefore, this invention not only broadens the application scope of Ginseng Strengthening Oral Liquid but also provides a safe and effective drug treatment approach for addressing age-related muscle function decline.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, and in particular relates to the application of ginseng tonic oral liquid in improving muscle function decline caused by aging. Background Technology

[0002] Skeletal muscle, as the largest organ system in the human body, is not only the main executive organ of the musculoskeletal system but also an important metabolic and endocrine organ, playing a crucial role in exercise, metabolism, and overall health. With age, skeletal muscle undergoes a series of complex structural and functional degenerative changes, involving multiple levels, from molecular and cellular alterations to macroscopic losses in muscle mass and strength. These changes collectively form a complex physiological network that directly affects muscle contractility, endurance, and fatigue characteristics, thereby influencing an older individual's daily activity levels, fall risk and fracture incidence, as well as exercise endurance and recovery capacity, ultimately impacting their quality of life and health status.

[0003] For example, at the microscopic level, aging muscles exhibit various structural changes, such as disordered myofibril arrangement, increased collagen deposition, and increased extracellular matrix accumulation. These changes collectively affect muscle contractile properties and mechanical performance. At the macroscopic level, decreased muscle strength is one of the most significant functional changes associated with aging, such as decreased grip strength. Grip strength is one of the most commonly used indicators for assessing muscle strength, and low grip strength is independently associated with various adverse outcomes, including all-cause mortality, cardiovascular disease, falls, fractures, and hospitalization. The causes of strength decline are multifaceted, with decreased muscle mass being the most direct cause.

[0004] Due to the lack of specific drugs, the approach to addressing the decline in skeletal muscle function and quality caused by aging usually combines exercise and nutrition, and drugs for this purpose remain one of the current research hotspots. Summary of the Invention

[0005] To address the above-mentioned technical problems, this invention provides the application of Ginseng Strengthening Oral Liquid in improving age-related muscle function decline. This invention is the first to discover that Ginseng Strengthening Oral Liquid can effectively improve age-related muscle strength decline, reduce collagen deposition, significantly improve muscle quality, and improve skeletal muscle damage, thereby effectively improving age-related skeletal muscle function decline. It can be used to develop effective prevention and intervention strategies for age-related decline in skeletal muscle function and quality.

[0006] To achieve the above-mentioned objectives, the present invention adopts the following technical solution:

[0007] This invention provides the application of ginseng tonic oral liquid in improving age-related muscle function decline.

[0008] Ginseng Strengthening Oral Liquid is a traditional Chinese medicine made primarily from ginseng. Its functions include nourishing yin and qi, strengthening the body's foundation, and treating various weakness symptoms caused by qi and yin deficiency, such as chronic cough (dry cough with little phlegm, dry throat and mouth), night sweats, shortness of breath, fatigue, palpitations, insomnia, lower back and knee weakness, and constipation. This invention, through research, has for the first time discovered that in aging mouse models, Ginseng Strengthening Oral Liquid can significantly improve grip strength, maintain or increase muscle mass, improve the integrity of muscle fiber structure, reduce collagen deposition, clear excess extracellular matrix, and make muscle structure denser. Furthermore, serum creatine kinase (CK) is a core enzyme in skeletal muscle cell metabolism. Serum CK levels are related to skeletal muscle mass and are often used as a clinical indicator to help assess the activity or metabolic state of muscle cells. When skeletal muscle cells are damaged, CK is released from the cells into the blood, leading to elevated serum CK levels. Creatine kinase isoenzyme MB (CK-MB) is an isoenzyme of the myocardium, but CK-MB may also be elevated in severe skeletal muscle injury or extreme fatigue. In sports medicine research, muscle fatigue (especially exercise-induced fatigue) leads to a transient increase in serum levels of skeletal muscle injury markers CK and CK-MB, reflecting micro-damage and metabolic disorders of muscle fibers under high-intensity activity. This invention experimentally demonstrates that Ginseng Strengthening Oral Liquid can reduce CK and CK-MB in aging mice. These results prove that Ginseng Strengthening Oral Liquid can effectively improve age-related muscle function decline.

[0009] Furthermore, while addressing age-related muscle function decline, Ginseng Strengthening Oral Liquid does not cause kidney damage, metabolic disorders, or abnormal weight of the liver and kidneys, demonstrating good systemic safety.

[0010] Currently, no studies have reported that Ginseng Strengthening Oral Liquid has an improving effect on age-related muscle function decline. The above research results of this invention broaden the application scope of Ginseng Strengthening Oral Liquid and provide a safe and effective drug treatment approach for addressing age-related muscle function decline.

[0011] Preferably, the muscle is skeletal muscle.

[0012] Preferably, the application is to improve grip.

[0013] Preferably, the application is to inhibit collagen deposition caused by aging.

[0014] Preferably, the application is for removing excess extracellular matrix.

[0015] Preferably, the application is for improving skeletal muscle damage caused by aging.

[0016] Preferably, the application is to reduce serum creatine kinase and / or creatine kinase isoenzyme MB.

[0017] The beneficial effects of this invention are as follows: This invention evaluated the effect of Ginseng Strengthening Oral Liquid on muscle function in D-galactose-induced aging mice through grip strength testing. It was the first time that Ginseng Strengthening Oral Liquid significantly improved the grip strength of aging mice, effectively alleviating the decline in muscle strength caused by aging. HE and Masson staining showed that the aging model mice exhibited disordered muscle structure and increased collagen deposition. After intervention with Ginseng Strengthening Oral Liquid, the intermuscular bundle gaps decreased in a dose-dependent manner, reaching a highly compact state at high doses. Serum CK and CK-MB levels were significantly elevated in the aging model mice, suggesting that aging can lead to skeletal muscle damage and metabolic disorders. Ginseng Strengthening Oral Liquid significantly reduced serum CK and CK-MB levels in aging mice, suggesting that it has a role in alleviating skeletal muscle damage, thereby improving fatigue by repairing muscle damage. Furthermore, while improving age-related muscle function, Ginseng Strengthening Oral Liquid did not cause kidney damage, metabolic disorders, or abnormal liver and kidney weight, demonstrating good systemic safety. Attached Figure Description

[0018] Figure 1 These are the gripping force test results in Embodiment 1 of the present invention; #### indicates p < 0.0001 compared with the normal group, ** indicates p < 0.05 compared with the model group, and **** indicates p < 0.0001 compared with the model group.

[0019] Figure 2 This is the result of the study on the ratio of muscle wet weight to body weight in Example 1 of the present invention; #### indicates p < 0.0001 compared with the normal group, **** indicates p < 0.0001 compared with the model group;

[0020] Figure 3 This is the HE staining result of the gastrocnemius muscle in Example 1 of the present invention;

[0021] Figure 4 This is the Masson staining result of the gastrocnemius muscle in Example 1 of the present invention;

[0022] Figure 5 These are the results of the investigation of serum biomarkers for skeletal muscle injury in Example 1 of this invention; * indicates p < 0.05 compared to the model group;

[0023] Figure 6 This is the result of the renal function safety evaluation in Example 1 of the present invention; * indicates p < 0.05 compared with the model group;

[0024] Figure 7 The safety evaluation results of the organ index in Embodiment 1 of the present invention. Detailed Implementation

[0025] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. All other embodiments obtained by those skilled in the art based on the implementation methods of this invention without inventive effort fall within the protection scope of this invention.

[0026] Aging can lead to a series of complex structural and functional degenerative changes in skeletal muscle, affecting muscle contractility, endurance, and fatigue characteristics, thereby impacting quality of life and causing health problems. Currently, the approach to addressing age-related decline in skeletal muscle function and quality typically combines exercise and nutrition, with a lack of specific medications.

[0027] Based on research on aging models, this invention is the first to discover that ginseng-strengthening oral liquid can effectively improve age-related muscle strength decline, reduce collagen deposition, significantly improve muscle mass, and improve skeletal muscle damage, thereby effectively improving the problem of age-related skeletal muscle function decline. Based on this, embodiments of this invention provide the application of ginseng-strengthening oral liquid in improving age-related muscle function decline.

[0028] The following detailed description, using specific examples, will further illustrate this point.

[0029] Unless otherwise specified, all other reagents and chemicals used in the following examples were obtained commercially. Unless otherwise specified, all experiments in the following examples employed conventional experimental methods in the art.

[0030] Example 1

[0031] This embodiment investigated the effects and safety of Renshen Guben Oral Liquid (RSGB) on skeletal muscle in an aging model mouse.

[0032] 1. Experimental Design and Grouping

[0033] 1.1 Laboratory Animals

[0034] SPF-grade healthy C57BL / 6 mice, male, weighing 25±2 g, 8 weeks old, purchased from Spf (Beijing) Biotechnology Co., Ltd., with animal license number SCXK(Beijing)2024-0001, were raised in the Experimental Animal Center of Tianjin University of Traditional Chinese Medicine at a room temperature of 20-25°C and a relative humidity of 40%-60%. All operations on experimental animals were strictly carried out in accordance with the standards of the Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (TCM-LAEC2025126L1965). The animal license number is SCXK(Beijing)2024-0001, and they were raised in the Experimental Animal Center of Tianjin University of Traditional Chinese Medicine at a room temperature of 20-25°C and a relative humidity of 40%-60%. All operations on experimental animals were strictly carried out in accordance with the standards of the Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (TCM-LAEC2025126L1965).

[0035] 1.2 Experimental drugs

[0036] Renshen Guben Oral Liquid, purchased from Lunan Hepu Pharmaceutical Co., Ltd., batch number 329230241.

[0037] VE solution: Take 1 commercially available vitamin E capsule (containing 0.1 g of vitamin E), cut it open and dissolve the contents in 5 ml of olive oil to prepare a solution with a concentration of 20 mg / mL for standby.

[0038] 1.2 Modeling and drug administration

[0039] The mice that had been adaptively fed for one week were randomly divided into a normal group (Control group), a model group (Model group), a low-dose RSGB group (L-RSGB group), a medium-dose RSGB group (M-RSGB group), a high-dose RSGB group (H-RSGB group), and a positive drug group (VE group), with 10 mice in each group. The mice in the Control group were subcutaneously injected with normal saline for 3 consecutive weeks, and the mice in other groups were subcutaneously injected with D-galactose (1 g / kg / d) for 3 consecutive weeks to establish a D-galactose-induced aging mouse model.

[0040] Starting from the day of modeling, the mice in each group were given gastric gavage once a day. The mice in the Control group and the Model group were gavaged with 0.5 mL of normal saline, the mice in the L-RSGB group, M-RSGB group, and H-RSGB group were gavaged with Renshen Guben Oral Liquid at doses of 2.5, 5, and 10 ml / kg / d respectively, and the mice in the VE group were gavaged with VE solution at a dose of 200 mg / kg / d. The weight of the mice was about 25 g, and the actual administration volume of each group was set according to this weight: L-RSGB group: 2.5×25×10 -3 =0.0625 (ml / d), M-RSGB group: 5×25×10 -3 =0.125 (ml / d), H-RSGB group: 10×25×10 -3=0.25 (ml / d), VE group: 200×25×10 -3 ÷20=0.25 (ml / d). Mice in each group were administered the drug for 3 weeks. Mouse weight was recorded daily during the administration period.

[0041] 1.3 Grasp Force Experiment

[0042] (1) Before the experiment, the mice were moved to the behavioral testing room to adapt to the environment for 30 minutes;

[0043] (2) Use a gripping force tester for testing;

[0044] (3) Grasp the mouse by the tail and make its forelimbs grip the metal grid of the gripping force tester, keeping the mouse's body in a horizontal position;

[0045] (4) Pull the mouse's tail backward at a constant speed until the mouse's forelimbs can no longer grip the grid and release it. Record the maximum gripping force value displayed by the instrument.

[0046] (5) Each mouse was measured 3 times consecutively, with an interval of 5 minutes between each measurement. The average value was taken as the gripping force value of the mouse (unit: g).

[0047] 1.4 Serum Sample Collection and Biochemical Indicator Detection

[0048] 1.4.1 Serum Sample Collection

[0049] (1) After the last administration, the mice were fasted but allowed to drink water for 12 h;

[0050] (2) Apical blood collection method: After anesthetizing the mice, the chest was opened to expose the heart. The blood collection needle was inserted into the apex of the left ventricle, and whole blood was gently aspirated and collected in an EP tube.

[0051] (3) Let stand at room temperature for 30 min, then centrifuge at 3000 rpm for 15 min at 4℃;

[0052] (4) Carefully extract the upper layer of serum, aliquot it and store it in a -80℃ refrigerator for later use.

[0053] 1.4.2 Serum Biochemical Indicators Detection

[0054] Serum samples frozen at -80℃ were thawed on ice and then processed strictly according to the instructions of the corresponding biochemical reagent kits. The levels of serum skeletal muscle injury markers CK and CK-MB, as well as the levels of renal function safety evaluation indicators, serum creatinine (CREA), blood urea nitrogen (UREA), uric acid (UA), albumin (ALB), serum calcium (Ca), and serum phosphorus (P) were measured using a fully automated biochemical analyzer.

[0055] 1.5 Safety evaluation of skeletal muscle histology and organ index

[0056] 1.5.1 Organ and Tissue Separation and Weighing

[0057] (1) After euthanizing the mice by taking blood from the apex of their hearts, the abdominal cavity was quickly opened and the intact liver and both kidneys were carefully separated.

[0058] (2) Rinse the separated liver and kidneys in pre-cooled saline to remove surface blood and connective tissue;

[0059] (3) After blotting off excess moisture from the organ surface with filter paper, immediately weigh the liver and kidneys using a precision electronic balance and record the results.

[0060] 1.5.2 Organ Index Calculation

[0061] Calculate the indices of each organ using the following formula:

[0062] Liver index = ×100% Liver Index

[0063] Kidney Index = ×100% Kidney Index

[0064] 1.6.1 Muscle wet weight to body weight ratio

[0065] After organ and tissue separation, the bilateral gastrocnemius, soleus, and quadriceps muscles of the mice were separated and weighed wetly. The relative muscle weight was calculated using the following formula: .

[0066] in: G represents the relative weight of muscle (%); G0 represents the wet weight of each muscle; G represents the body weight of the mouse (g).

[0067] 1.6.2 Observation of skeletal muscle tissue structure (HE staining)

[0068] Mouse gastrocnemius muscle tissue was collected and fixed with 4% paraformaldehyde. After paraffin embedding and sectioning, routine hematoxylin-eosin staining (HE) was performed. The middle part of each slide was observed and analyzed under a 20x digital slide scanner.

[0069] 1.6.3 Assessment of skeletal muscle fibrosis (Masson staining)

[0070] Mouse gastrocnemius muscle tissue was collected and fixed with 4% paraformaldehyde. After paraffin embedding and sectioning, routine Masson staining was performed, and the middle part of each slide was observed and analyzed under a 20x digital slide scanner.

[0071] 1.7 Statistical Analysis

[0072] All data in this study are expressed as mean ± standard deviation ( ). Mean ± standard deviation (S) indicates statistical significance. GraphPad Prism 10.1.2 statistical analysis software was used for analysis. The t-test was used for comparisons between the two groups, and P < 0.05 was considered statistically significant.

[0073] 2. Results

[0074] 2.1 Grasp Force Experiment

[0075] A significant decrease in grip strength is a typical behavioral manifestation of age-related fatigue. Grip strength test results ( Figure 1 The results showed that, compared with the model group, all dosage groups (low, medium, and high) of Ginseng Strengthening Oral Liquid significantly improved the grip strength of mice (p < 0.05), with the high-dose group showing the most significant effect, and the grip strength value approaching that of the normal control group. Furthermore, the drug effect exhibited a clear dose-dependent relationship (H-RSGB > M-RSGB > L-RSGB). These results indicate that Ginseng Strengthening Oral Liquid can significantly improve the grip strength of aging mice and effectively alleviate age-related muscle strength decline.

[0076] 2.2 Results of skeletal muscle histological examination

[0077] 2.2.1 Muscle wet weight to body weight ratio

[0078] The results are as follows Figure 2 As shown, compared with the control group, the relative muscle weight of mice in the model group was slightly increased, suggesting a possible model adaptation or edema response. Muscle tissue may have increased "wet weight" due to factors such as edema and fibrosis. After intervention with Ginseng Guben Oral Liquid, the relative muscle weight of each dosage group (L-RSGB, M-RSGB, H-RSGB) decreased compared with the model group, suggesting that Ginseng Guben Oral Liquid may maintain or improve muscle quality by improving muscle metabolism, eliminating edema, and reducing collagen deposition.

[0079] 2.2.2 Observation of skeletal muscle tissue structure

[0080] The results are as follows Figure 3 As shown, compared with the Control group, the gastrocnemius muscle tissue of the Model group mice exhibited disordered structure, with muscle bundles of varying sizes and loose arrangement. Scattered inflammatory cell infiltration was observed in the interstitium, and inflammatory cells were seen surrounding some small blood vessels. This suggests that D-galactose successfully induced age-related muscle pathological changes. After VE intervention, muscle pathology was partially improved, with muscle bundle arrangement more orderly than in the Model group. The uneven size of muscle fibers still existed, but to a lesser degree than in the Model group, and interstitial inflammatory cell infiltration was reduced. After intervention with Ginseng Strengthening Oral Liquid (RSGB), compared with the Model group, the integrity of fiber structure gradually recovered in each RSGB dose group, and muscle pathology improved in a dose-dependent manner.

[0081] 2.2.3 Assessment of the degree of skeletal muscle fibrosis

[0082] The results are as follows Figure 4 As shown, D-galactose-induced gastrocnemius muscle in aging mice exhibited a "diffuse imbalance" state and structural abnormalities. Masson staining revealed that, compared to the Control group, the perimysium of the gastrocnemius muscle in the Model group mice was significantly thickened, with blue collagen fibers surrounding and separating the muscle bundles, resulting in widened and loosely arranged intermuscular bundle spaces (arrows indicated). After intervention with ginseng-based oral liquid, the intermuscular bundle spaces decreased in a dose-dependent manner, showing a clear dose-dependent improvement; the high-dose group reached a highly compact state, including signs of active angiogenesis and regeneration, while the low-dose group also showed suspected angiogenesis, and the blue collagen fibers also became thinner. The positive control group (VE) also showed a stabilizing effect.

[0083] Both HE and Masson staining results demonstrated that Ginseng Strengthening Oral Liquid can reduce collagen deposition, remove excess extracellular matrix, and make muscle structure denser. These results further confirm that Ginseng Strengthening Oral Liquid can improve muscle quality, thereby alleviating the aforementioned pathological changes in skeletal muscle of aging mice and relieving age-related fatigue-like symptoms.

[0084] 2.2.4 Serum markers of skeletal muscle injury

[0085] The results are as follows Figure 5 As shown, compared with the Control group, the serum creatine kinase (CK) and creatine kinase isoenzyme MB (CK-MB) levels in the Model group were significantly elevated, with a significant difference in the degree of CK-MB elevation. This suggests that D-galactose-induced aging mice have significant skeletal muscle damage, possibly accompanied by some degree of myocardial metabolic abnormalities. Ginseng tonic oral liquid effectively reduced the elevated CK and CK-MB levels in the serum of aging mice, and the effect on reducing CK-MB levels showed a significant difference.

[0086] 2.2.5 Evaluation of renal function safety

[0087] The results are as follows Figure 6 As shown, compared with the Control group, the Model group had significantly higher serum creatinine (CREA-S) and blood urea nitrogen (UREA), and the degree of increase in CREA-S was significantly different, indicating a renal burden. After intervention with Ginseng Guben Oral Liquid, the functional indicators of CREA-S and UREA decreased relative to the Model group, confirming the systemic safety of the drug and indicating that Ginseng Guben Oral Liquid can improve abnormal renal function without nephrotoxicity. There were no significant changes in serum uric acid (UA), albumin (ALB), serum calcium (Ca), and serum phosphorus (P) levels among the groups, suggesting that Ginseng Guben Oral Liquid did not affect purine metabolism, protein synthesis, or electrolyte homeostasis.

[0088] In summary, Ginseng Strengthening Oral Liquid addresses age-related muscle function decline without causing kidney damage or metabolic disorders, demonstrating good systemic safety.

[0089] 2.2.6 Safety evaluation of organ index

[0090] There were no significant changes in liver index (liver weight / body weight) and kidney index (kidney weight / body weight) in any group of mice, suggesting that ginseng tonic oral liquid did not cause abnormal weight of liver and kidney organs at the experimental dose, further supporting its systemic safety.

[0091] 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, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Application of Ginseng Strengthening Oral Liquid in Improving Muscle Function Decline Due to Age.

2. The application according to claim 1, characterized in that, The muscle in question is skeletal muscle.

3. The application according to claim 1 or 2, characterized in that, The application described is to improve grip.

4. The application according to claim 1 or 2, characterized in that, The application is to inhibit collagen deposition caused by aging.

5. The application according to claim 1 or 2, characterized in that, The application is to remove excess extracellular matrix.

6. The application according to claim 1 or 2, characterized in that, The application is for improving skeletal muscle damage caused by aging.

7. The application according to claim 1 or 2, characterized in that, The application is to reduce serum creatine kinase and / or creatine kinase isoenzyme MB.

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