Application of tortoise and deer preparation for tonifying kidney in preparation of drugs for resisting aging and / or delaying senile muscle attenuation syndrome

Abstract

The application discloses application of a turtle and deer kidney tonifying preparation in preparation of anti-aging and / or delaying senile muscle attenuation syndrome drugs. Through an old beagle dog model experiment, it is shown that after continuously taking the turtle and deer kidney tonifying pills for 60 days, not only can the in-vivo tumor necrosis factor-alpha and oxidative stress product malondialdehyde levels be significantly reduced, the muscle mass and strength and muscle mitochondria level can be improved, the function of improving tibia and bone density can be played, but also the old animal cognitive behavior function, ovary function, bladder function and gastrocnemius function can be improved, and the overall anti-aging and delaying senile muscle attenuation syndrome effect can be played, which may be related to the fact that the turtle and deer kidney tonifying preparation can down-regulate the expression of muscle aging related genes p16, p27 and Tp53, and improve aging. A pre-experiment shows that other turtle and deer kidney tonifying oral preparations also have the same or similar effects.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a new pharmaceutical use of a traditional Chinese medicine composition, a tortoise and deer kidney-tonifying preparation, and particularly to its application in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome. Technical Background With the increasing aging of the global population, aging and related degenerative diseases have become a significant public health issue. Sarcopenia, also known as age-related muscle loss syndrome, is a progressive and widespread degenerative syndrome associated with aging, characterized by a decrease in skeletal muscle mass, strength, and function. This disease severely impacts the quality of life of older adults, increasing the risk of falls, fractures, disability, and death. Currently, clinical interventions for senile sarcopenia are limited, primarily including nutritional supplementation, physical exercise, and a few drugs still under investigation. There is a lack of treatments with proven efficacy and high safety profiles.

[0002] The aging process is often accompanied by systemic chronic low-grade inflammation, oxidative stress damage, mitochondrial dysfunction, and abnormal expression of cellular senescence-related genes. In muscle tissue, pro-inflammatory factors such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) are elevated, antioxidant capacity is decreased, malondialdehyde (MDA), a lipid peroxidation product, accumulates, and the ratio of oxidized nicotinamide adenine dinucleotide to reduced nicotinamide adenine dinucleotide (NAD), a key molecule in mitochondrial energy metabolism, is also affected. + The levels of NADH and adenosine triphosphate (ATP) decrease, while the expression of aging markers such as cell cycle inhibitors p16, p27, Tp53, and Cdkn1a is upregulated, ultimately leading to muscle fiber atrophy and a decline in muscle regeneration and repair capabilities.

[0003] Guilu Bushen Wan (including Guilu Bushen Wan) is a traditional Chinese medicine preparation made from tortoise shell glue, deer antler glue, salt-processed dodder seed, epimedium, dipsacus root, cynomorium, cibotium, jujube seed, prepared he shou wu, prepared licorice root, tangerine peel, prepared rehmannia root, rosehip, prepared astragalus root, yam, and raspberry, among other Chinese herbs. It has the effects of tonifying the kidneys and strengthening yang, replenishing qi and blood, and strengthening muscles and bones. Clinically, it is mainly used for symptoms such as physical weakness, mental fatigue, soreness in the lower back and legs, dizziness, frequent urination at night, forgetfulness, and insomnia. However, there are currently no research or application reports on Guilu Bushen Wan or other oral preparations with the same formula in anti-aging and delaying the onset of age-related sarcopenia syndrome. Summary of the Invention

[0004] The purpose of this invention is to provide a new use for the tortoise and deer kidney-tonifying preparation, and its application in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome.

[0005] The present invention, through experiments with an aged Beagle dog model, proves that Guilu Bushen Pills can significantly improve a number of aging- and sarcopenia-related indicators in aged dogs, which is specifically reflected as follows: 1. Improving cognitive and behavioral functions: significantly increasing the exploration duration and times of aged animals to novel objects, improving cognitive learning ability and exploration desire; shortening the adaptation time to new environments and ameliorating social interaction disorders, which indicates that it has the effect of improving cognitive and behavioral decline in aged dogs.

[0006] 2. Inhibiting inflammatory response: low-dose and high-dose administration can significantly reduce the level of TNF-α in serum, and high-dose administration can significantly reduce the level of IL-6 in serum, thus inhibiting the inflammatory response in aged dogs.

[0007] 3. Improving mitochondrial function: significantly reducing the level of H2O2 in the body, increasing the NAD + + / NADH ratio and ATP content, and significantly increasing the number of mitochondria in gastrocnemius muscle, which suggests that it can improve energy metabolism and mitochondrial function.

[0008] 4. Delaying sarcopenia: the high-dose group significantly increases the gastrocnemius organ coefficient (wet muscle weight / body weight), increases the content of serum myoglobin (MB) and the level of creatine kinase (CK) in gastrocnemius muscle, delays muscle tissue atrophy and improves muscle quality. Meanwhile, the peak tension is significantly increased in muscle strength assessment.

[0009] 5. Inhibiting the expression of muscle aging genes: the high-dose group significantly down-regulates the expression of aging-related genes p16, p27 and Tp53 in gastrocnemius muscle, which suggests that it can delay the process of muscle aging and improve muscle regeneration ability.

[0010] 6. Improving ovarian and bladder functions: the levels of serum estradiol (E2) and anti-Müllerian hormone (AMH) are increased to a certain extent, indicating that Guilu Bushen preparation has the effects of improving ovarian reserve, delaying ovarian aging and enhancing ovarian endocrine function; histopathology shows that the degenerative changes of the ovary are improved.

[0011] 7. Increasing bone mineral density: can delay the decline of the skeletal system and increase bone mineral density to a certain extent.

[0012] 8. Effects on liver, gallbladder and kidney functions: blood biochemical examination shows that Guilu Bushen preparation has no influence on liver, gallbladder and kidney functions.

[0013] 9. Delaying the decline of bladder and gastrocnemius muscle: can delay the age-related degenerative changes of bladder and gastrocnemius muscle and improve the functions of bladder and gastrocnemius muscle to a certain extent.

[0014] Therefore, Guilu Bushen preparation can exert an anti-aging effect at the overall level, and has a significant delaying effect especially on age-related sarcopenia, providing a new safe and effective therapeutic option for clinical practice.

[0015] In the present invention, the clinical daily dosage of Guilu Bushen Pills or other oral preparations calculated as the amount of raw medicinal material is 18g / day (for an adult weighing 70 kg). The animal administration dosage is converted based on body surface area, the low dose is 0.48 g / kg / day, and the high dose is 0.96 g / kg / day, which is equivalent to 1-2 times of the intended clinical dose (by body surface area method) or 1.8-3.7 times (by body weight method). The administration route is oral gavage, and the administration is continued for 60 days. The dosage range has no obvious adverse effects on liver and kidney functions and blood glucose, with good safety Other Guilu Bushen preparations can be other common oral dosage forms such as oral liquid, tablets, capsules, granules, dropping pills, and medicinal pastes. According to preliminary pre-experiments, other oral dosage forms have the same or similar efficacy and toxicity test results as Guilu Bushen Pills. Description of Drawings

[0016] Figure 1 is the result graph of the number of mitochondria in the gastrocnemius of the young dog negative control group, the aged dog negative control group, the low-dose Guilu Bushen Pill group, and the high-dose Guilu Bushen Pill group in the test of the present invention.

[0017] Figure 2 is the histopathological examination result graphs of ovary tissue, bladder tissue and gastrocnemius tissue of the young dog negative control group, the aged dog negative control group, the low-dose Guilu Bushen Pill group, and the high-dose Guilu Bushen Pill group in the test of the present invention. Detailed Description of Embodiments

[0018] The present invention is further illustrated by the following examples, but the protection scope of the present invention is not limited thereto.

[0019] Example 1: Safety Evaluation of Guilu Bushen Pills 1. Experimental Animals and Grouping Eighteen female aged Beagle dogs that passed quarantine and were older than 96 months of age were selected, they were randomly divided into the aged dog negative control group, the low-dose Guilu Bushen Pill group, and the high-dose Guilu Bushen Pill group, with 6 dogs in each group. Another 6 young female Beagle dogs aged 12-15 months were selected as the young dog negative control group.

[0020] 2. Administration Treatment The low-dose and high-dose Guilu Bushen Pill groups were orally administered Guilu Bushen Pill capsules at dosages of 0.48 g / kg and 0.96 g / kg respectively, once a day for consecutive 60 days. The young dog negative control group and the aged dog negative control group were given empty gelatin capsules.

[0021] 3. Safety Evaluation Liver, gallbladder, and kidney function tests: Clinical examinations include blood biochemistry tests to measure the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), total bile acids (TBA), total bilirubin (TBIL), direct bilirubin (DBIL), gamma-glutamyl transferase (GGT), urea (UREA), and creatinine (CREA). These tests can reflect changes in liver, gallbladder, and kidney function to some extent. Sixty days after drug administration, venous blood is collected, and serum is separated for blood biochemistry analysis.

[0022] Table 1. Effects of the test substance on liver and biliary function and blood biochemical parameters in animals after 60 days of administration (x±s)

[0023] Table 2. Effects of the test substance on bile, kidney function, and blood biochemical parameters in animals after 60 days of administration (x±s)

[0024] As shown in Tables 1 and 2, no significant changes were observed in the blood biochemical indicators of liver function such as ALT, AST, ALB, and LDH after administration, indicating that Guilu Bushen Wan has no effect on liver function; no significant changes were observed in the blood biochemical indicators of gallbladder function such as ALP, TBA, TBIL, DBIL, and GGT, indicating that Guilu Bushen Wan has no effect on gallbladder function; and no significant changes were observed in the blood biochemical indicators of kidney function such as UREA and CREA, indicating that Guilu Bushen Wan has no effect on kidney function.

[0025] Example 2: Efficacy test of Guilu Bushen Wan (a traditional Chinese medicine formula) in anti-aging and delaying of senile sarcopenia in aged Beagle dogs. Treatment efficacy testing (1) New and old object recognition test and dog-dog interaction test: New and old object recognition test and dog-dog interaction test are commonly used detection methods in animal cognition and behavior. Specifically, new and old object recognition: In an open space, a familiar object (old object) is placed in the dog's field, and a new object that is similar in color and size to the familiar toy but different in shape is placed in the field. Record the number of times and duration of the animal explores the new and old objects within 3 minutes and calculate the percentage. Dog-dog interaction test: The dog-dog interaction test is conducted using a 3-chamber test; the test is carried out in a test environment containing three chambers, namely the autonomous chamber, the unfamiliar dog chamber 1 and the unfamiliar dog chamber 2; in the adaptation phase, the dog is placed in the autonomous chamber and observed for 10 minutes; in the second phase, unfamiliar dog 1 is placed in the dog cage in unfamiliar dog chamber 1 and the behavior of the test dog is observed for 10 minutes; in the third phase, unfamiliar dog 2 is placed in the dog cage in unfamiliar dog chamber 2 and the behavior of the test dog is observed for 10 minutes. Record the total number of times and the total duration of the test dog entering unfamiliar dog chamber 1 and unfamiliar dog chamber 2 in the second and third phases.

[0026] Table 3. Effects of the test substance on the ability of animals to recognize new and old objects after 60 days of administration (x±s)

[0027] Compared with the negative control group of young dogs, # express p≤ 0.05, ## express p ≤0.01. Compared with the negative control group of aged dogs, * express p≤ 0.05, ** express p ≤0.01.

[0028] As shown in Table 3, compared with the negative control group of aged dogs, the low and high dose groups of Guilu Bushen Pills increased the percentage of time and number of times the aged animals explored new objects (p≤0.05), indicating that Guilu Bushen Pills of the present invention can improve the decline in cognitive learning ability of aged animals after use.

[0029] Table 4. Effects of the test substance on animal cognition and environmental adaptability after 60 days of administration (x±s)

[0030] Compared with the negative control group of young dogs, # express p≤ 0.05, ## express p ≤0.01. Compared with the negative control group of aged dogs, * express p≤ 0.05, ** express p ≤0.01.

[0031] Table 5. Effects of the test substance on cognitive and social abilities in animals after 60 days of administration (x±s)

[0032] Compared with the negative control group of young dogs, # express p≤ 0.05, ## express p ≤0.01. Compared with the negative control group of aged dogs, * express p≤ 0.05, ** express p ≤0.01.

[0033] As shown in Tables 4 and 5, high doses of Guilu Bushen Wan can reduce the time animals take to adapt to unfamiliar environments to a certain extent, suggesting that Guilu Bushen Wan can improve the adaptability of older animals to unfamiliar environments to a certain extent. In the second stage, low doses of Guilu Bushen Wan can increase the interaction time between older animals and unfamiliar dog 1 to a certain extent, and reduce the time spent exploring the empty cage. In the third stage, low and high doses of Guilu Bushen Wan can increase the interaction time between older animals and unfamiliar dog 2 to a certain extent, and reduce the interaction time with unfamiliar dog 1. This indicates that Guilu Bushen Wan of the present invention can improve social impairment and cognitive and social abilities to a certain extent after use.

[0034] (2) Detection of inflammatory factors and oxidative stress response: The levels of TNF-α and IL-6 in serum were detected using commercial kits.

[0035] Table 6. Effects of test substance administration on serum inflammatory factor levels in animals after 60 days (x±s)

[0036] Compared with the negative control group of young dogs, # express p≤ 0.05, ## express p ≤0.01. Compared with the negative control group of aged dogs, * express p≤ 0.05, ** express p ≤0.01.

[0037] Table 6 shows that both low and high doses of Guilu Bushen Wan significantly reduced serum TNF-α levels (p≤0.05), and high doses of Guilu Bushen Wan significantly reduced serum IL-6 levels (p≤0.05). This indicates that Guilu Bushen Wan of the present invention can improve the inflammatory response in aging bodies after use.

[0038] (3) Blood and gastrocnemius muscle mitochondrial function tests: H2O2 and NAD in serum were detected using commercially available kits. + NADH, ATP levels, and NAD in the gastrocnemius muscle + / NADH levels; simultaneously, the number of mitochondria in the gastrocnemius muscle was detected by electron microscopy using the following method: Gastrocnemius muscle tissue was fixed in electron microscopy fixative at room temperature for 2 hours, rinsed with phosphate buffer, fixed with 1% osmium tetroxide at room temperature for 2 hours, and rinsed again; subsequently, it was dehydrated sequentially with a gradient of 30%-100% alcohol, then permeated stepwise with a mixture of acetone and epoxy resin, and finally embedded in epoxy resin and polymerized at 60°C for 48 hours; sections were cut into 80-100 nm sections using an ultramicrotome, stained with uranium lead double staining, and observed and images were acquired under a scanning electron microscope to calculate the number of mitochondria in each image. These indicators are commonly used to detect mitochondrial function and muscle energy metabolism.

[0039] Appendix Figure 1 are the result graphs of the number of mitochondria in the gastrocnemius of the young dog negative control group, the aged dog negative control group, the low-dose Guilu Bushen Wan group and the high-dose Guilu Bushen Wan group in the experiment of the present invention.

[0040] Table 7 Effects of test substance on animal blood oxidative stress / energy metabolism indicators, number of gastrocnemius mitochondria and NAD+ / NADH after 60 days of administration (x±s)

[0041] Compared with the young dog negative control group, # indicates p≤ p < 0.05, ## indicates p p ≤ 0.01. Compared with the aged dog negative control group, * indicates p≤ p < 0.05, ** indicates p p ≤ 0.01.

[0042] It can be seen from Table 7 and Appendix Figure 1 that the high-dose Guilu Bushen Wan can not only significantly down-regulate the level of H2O2 (p ≤ 0.05) in blood and significantly increase the number of mitochondria in gastrocnemius (p ≤ 0.05), but also up-regulate the level of NAD + + / NADH in blood and gastrocnemius. It indicates that the use of Guilu Bushen Wan in the present invention can improve the oxidative stress response of gastrocnemius mitochondria, increase the number of mitochondria in muscle tissue, and improve the energy conversion function of mitochondria.

[0043] (4) Detection of gastrocnemius organ coefficient and muscle strength: The gastrocnemius organ coefficient is commonly used to evaluate the calf muscle mass of animals. The gastrocnemius weight is weighed to calculate the gastrocnemius organ coefficient (gastrocnemius organ coefficient = gastrocnemius weight / animal body weight * 100%), and the peak tension of the animal is detected at the same time: a tension gauge is used to detect the strength level of the dog's muscle groups, which can be used to evaluate the real-time strength of the animal's muscle groups; a commercial kit is used to detect the levels of CK in gastrocnemius and MB in serum.

[0044] Table 8 Effects of test substance on animal gastrocnemius organ coefficient and muscle strength after 60 days of administration (x±s)

[0045] Compared with the young dog negative control group, # indicates p≤ p < 0.05, ## indicates p p ≤ 0.01. Compared with the aged dog negative control group, * indicates p≤ p < 0.05,** express p ≤0.01.

[0046] As shown in Table 8, high doses of Guilu Bushen Wan significantly increased the number of visceral systems, peak tensile strength, and CK level in the gastrocnemius muscle (p≤0.05), while also increasing MB levels towards the negative control group in young dogs. This indicates that using Guilu Bushen Wan in this invention can improve muscle loss and strength decline, slow down the rate of muscle atrophy, and increase the rate of muscle synthesis.

[0047] (5) Inhibition of muscle aging gene expression: Fresh or frozen animal tissue was minced and homogenized with TRIzon Reagent (TRIzon total RNA extraction reagent) and placed at room temperature for 5 minutes; chloroform was added in proportion and shaken, and after centrifugation, the upper aqueous phase was collected, and an equal volume of 70% ethanol was added and mixed. The mixture was then transferred to an adsorption column and washed sequentially with Buffer RW1 (RW1 washing buffer) and Buffer RW2 (RW2 washing buffer). After centrifugation and drying, RNase-free water was added to elute and obtain total ribonucleic acid (RNA); 500 ng of RNA was used to remove genomic deoxyribonucleic acid (DNA) with gDNA Erase and then reverse transcribed to synthesize complementary DNA (cDNA); then 5 ng of cDNA was used as a template, and primers and SYBR Green Supermix were added to perform real-time quantitative PCR (95℃ 3min; 40 cycles: 95℃ 15s, 60℃ 30s, 72℃) The expression levels of target genes were calculated using a relative quantification method with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an internal control (20s). The expression of messenger RNA (mRNA) of aging-related markers p16, p27, and Tp53 in muscle was detected by reverse transcription polymerase chain reaction (RT-PCR).

[0048] Table 9. Effects of the test substance on aging-related markers in the gastrocnemius muscle of animals after 60 days of administration (x±s)

[0049] Compared with the negative control group of young dogs, # express p≤ 0.05, ## express p ≤0.01. Compared with the negative control group of aged dogs, * express p≤ 0.05, ** express p ≤0.01.

[0050] It can be seen from Table 9 that the high-dose Guilu Bushen Pills can significantly down-regulate the mRNA expression levels of aging-related markers p16 (p≤0.05), p27 (p≤0.01) and Tp53 (p≤0.01) in gastrocnemius muscle, indicating that the use of the Guilu Bushen Pills of the present invention can reduce the expression level of muscle aging genes, improve muscle atrophy caused by aging, slow down the rate of muscle aging, and delay the decline of muscle function.

[0051] (6) Ovarian function: Detection of serum E2 and AMH levels by kits and histopathological examination of ovary are commonly used methods to evaluate ovarian function.

[0052] Appendix Figure 2 shows the histopathological examination results of ovarian tissue, bladder tissue and gastrocnemius muscle tissue of young dog negative control group, aged dog negative control group, low-dose Guilu Bushen Pills group and high-dose Guilu Bushen Pills group in the test of the present invention.

[0053] Table 10 Effect of the test substance on animal ovarian function after 60 days of administration (x±s)

[0054] Compared with the young dog negative control group, # indicates p≤ 0.05, ## indicates p ≤0.01. Compared with the aged dog negative control group, * indicates p≤ 0.05, ** indicates p ≤0.01.

[0055] It can be seen from Table 10 that compared with the aged dog negative control group, the serum levels of E2 and AMH in the high-dose Guilu Bushen Pills group are increased, indicating that Guilu Bushen Pills has the effects of improving ovarian reserve, delaying ovarian aging, and enhancing ovarian endocrine function. Meanwhile, the appendix Figure 2 also shows that ovarian degenerative changes are improved under high-dose Guilu Bushen Pills, indicating that the Guilu Bushen Pills of the present invention can improve ovarian function degeneration after administration.

[0056] (7) Bone mineral density test: Bone mineral density refers to the content of minerals per unit area, which is clinically used as an indicator to reflect the degree of osteoporosis. The process of bone mineral density test is as follows: after 60 days of administration, the left lower legs of dogs from different experimental groups are taken, and an X-ray bone mineral density analyzer is used to test the bone mineral density of the femur of dogs.

[0057] Table 11 Effect of the test substance on animal bone mineral density after 60 days of administration (x±s)

[0058] Compared with the young dog negative control group,# express p≤ 0.05, ## express p ≤0.01. Compared with the negative control group of aged dogs, * express p≤ 0.05, ** express p ≤0.01.

[0059] As shown in Table 11, compared with the negative control group of elderly dogs, the bone mineral density of the femur in the high-dose group of Guilu Bushen Pills was significantly increased (p≤0.05), indicating that Guilu Bushen Pills in this invention have a good effect on improving osteoporosis after use.

[0060] (8) Histopathological examination of the bladder, ovary, and gastrocnemius muscle: Histopathological examination can further confirm the degenerative lesions of the bladder, ovary, and gastrocnemius muscle, and can be used to assess the function of the bladder, ovary, and gastrocnemius muscle. Histopathological examination is performed as follows: The target organ is fixed in 4% paraformaldehyde, routinely embedded in paraffin, and then prepared into 2-3 µm sections. After hematoxylin-eosin staining (HE staining), histopathological examination is performed: observe the epithelial mucosa layer, interstitial inflammatory cell infiltration, fibrous tissue between the lamina propria and muscle layer of the bladder, and the bladder wall; observe the number of primordial and primary follicles of the ovary, interstitial inflammatory cell infiltration, and atrophy and sclerosis; observe whether the gastrocnemius muscle is atrophied and the condition of muscle fiber lesions.

[0061] As attached Figure 2 As shown, the negative control group of aged dogs exhibited significant age-related degenerative changes in the gastrocnemius muscle, bladder, and ovaries, manifested as moderate muscle atrophy and mild vacuolation of the gastrocnemius muscle, moderate increase in fibrous tissue between the lamina propria and muscle layer of the bladder mucosa and mild thinning of the bladder wall, and moderate reduction and moderate atrophy and sclerosis of primordial and primary follicles in the ovaries, consistent with the aging characteristics of tissues and organs in aged animals. The low- and high-dose groups of Guilu Bushen Wan (a traditional Chinese medicine formula) improved these age-related pathological changes to varying degrees, showing a certain dose-related correlation. The high-dose group showed a more significant improvement, significantly alleviating gastrocnemius muscle atrophy and vacuolation, reducing bladder fibrous tissue hyperplasia and wall thinning, and improving the reduction and atrophy of ovarian follicles. Furthermore, no significant pathological damage related to the test substance was observed in any of the dose groups. Guilu Bushen Wan can effectively improve the age-related degenerative changes in the gastrocnemius muscle, bladder, and ovaries of aged Beagle dogs. The results indicate that Guilu Bushen Wan of this invention can improve bladder function and delay gastrocnemius muscle atrophy after use.

[0062] Preliminary experimental results show that other oral preparations of the present invention have the same or similar pharmacodynamic and toxicological results as Guilu Bushen Pills.

Claims

1. Application of tortoise and deer kidney-tonifying preparations in the preparation of anti-aging and / or age-related sarcopenia syndrome drugs.

2. The application of the tortoise and deer kidney-tonifying preparation according to claim 1 in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome, characterized in that... Application of tortoise and deer kidney-tonifying preparations in the preparation of drugs to improve cognitive and behavioral decline.

3. The application of the tortoise and deer kidney-tonifying preparation according to claim 1 in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome, characterized in that... The application of tortoise and deer kidney-tonifying preparations in the preparation of drugs that reduce serum TNF-α and IL-6 levels and inhibit inflammatory responses in the body.

4. The application of the tortoise and deer kidney-tonifying preparation according to claim 1 in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome, characterized in that... The preparation of the tortoise and deer kidney-tonifying preparation significantly reduced the H2O2 level in whole blood and gastrocnemius muscle and increased NAD. + The application of drugs that increase the NADH ratio and ATP content, increase the number of mitochondria in the gastrocnemius muscle, and improve energy metabolism and mitochondrial function.

5. The application of the tortoise-deer kidney-tonifying preparation according to claim 1 in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome, characterized in that... The application of tortoise and deer kidney-tonifying preparations in the preparation of drugs that increase the number of visceral systems in the gastrocnemius muscle, increase serum myoglobin content and gastrocnemius muscle creatine kinase level, delay muscle tissue atrophy, improve muscle quality, and enhance peak tensile strength in muscle strength assessment.

6. The application of the tortoise and deer kidney-tonifying preparation according to claim 1 in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome, characterized in that... Application of tortoise and deer kidney-tonifying preparation in the preparation of drugs that downregulate the expression of aging-related genes p16, p27, and Tp53 in the gastrocnemius muscle.

7. The application of the tortoise and deer kidney-tonifying preparation according to claim 1 in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome, characterized in that... Application of tortoise and deer kidney-tonifying preparations in the preparation of drugs that increase serum E2 and AMH levels and improve ovarian degenerative changes.

8. The application of the tortoise and deer kidney-tonifying preparation according to claim 1 in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome, characterized in that... Application of tortoise and deer kidney-tonifying preparations in the preparation of drugs to improve bone density.

9. The application of the tortoise and deer kidney-tonifying preparation according to claim 1 in the preparation of drugs for anti-aging and / or delaying senile sarcopenia syndrome, characterized in that... Application of tortoise and deer kidney-tonifying preparations in the preparation of drugs that delay age-related degenerative changes in the bladder and gastrocnemius muscles and improve the function of the bladder and gastrocnemius muscles.

10. The tortoise and deer kidney-tonifying preparation according to any one of claims 1 to 9, characterized in that, The pharmaceutical preparation may be replaced with an oral preparation of one of the following with the same raw material ratio: oral liquid, tablet, capsule, granule, drop pill or decoction.

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