Application of Jintiange capsule in delaying skeletal muscle senescence

Abstract

The invention provides application of Jintiange capsules in delaying skeletal muscle senescence. The invention has the beneficial effects that the Jintiange capsule simulates the traditional Chinese medicine tiger bone powder through the bionic tiger bone powder, effectively delays skeletal muscle senescence, and can be directly applied clinically by using the medicine verified by clinical examination and approval. The Jintiange capsule is composed of various animal bones and has the characteristics of multiple components and multiple target points, so that no serious safety problem is found in clinical application of the Jintiange capsule. In addition, the old people have good compliance after taking the Jintiange capsule, and the long-term taking of the Jintiange capsule has feasibility for delaying skeletal muscle aging.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to the use of Jintiange capsules (main component is artificial tiger bone powder) in delaying skeletal muscle aging. BACKGROUND

[0002] Muscle aging refers to a syndrome of progressive, systemic skeletal muscle mass reduction, muscle strength decline and functional decline with age increase, which is closely related to falls, fractures, metabolic disorders and increased mortality. Muscle aging is often accompanied by adverse physiological and physical changes, which can lead to increased disease incidence, decreased quality of life, increased mortality and increased medical costs. At present, there is no clinically approved drug specifically for muscle aging worldwide. Existing researches are mostly limited to phase II or phase III trials, with limited improvement in efficacy endpoints, and lack of long-term safety data.

[0003] Resistance training and high protein supplementation are considered as the 'gold standard', but the compliance of the elderly population is poor, especially those with cognitive impairment or bone and joint diseases. Even if exercise and lifestyle interventions are strictly implemented, about 20-30% of the elderly individuals still cannot prevent the rapid occurrence of muscle aging [1] . In addition, hormone therapy is not widely used in clinical practice due to its poor safety, single target and poor benefit-risk ratio.

[0004] Jintiange capsules (JTG) are a marketed traditional Chinese medicine (Guo Yao Zheng Zhi Z20030080), which is a biomimetic tiger bone powder similar to natural tiger bone components, rich in bioactive peptides, proteins, amino acids and minerals, and is made of the bones of various domestic animals, including pigs, goats and deer [2] . The indications recorded in the instruction manual are “back pain, soreness of the waist and knees, and osteoporosis”, and a large number of experimental studies and large-scale clinical trials have confirmed that JTG has significant effects in relieving osteoporosis, improving bone microstructure and enhancing the bone biomechanical properties of ovariectomized rats. However, no published research has revealed the direct regulatory effect of Jintiange capsules on muscle aging and the molecular mechanism. SUMMARY

[0005] The purpose of the present application is to study the effect of Jintiange capsules in delaying skeletal muscle aging.

[0006] To achieve the above-mentioned purpose, the present application provides the use of Jintiange capsules in delaying skeletal muscle aging.

[0007] The beneficial effects of the application are: the Jintiange capsule simulates traditional Chinese medicine tiger bone powder by imitating tiger bone powder, effectively delays skeletal muscle aging, and uses clinically approved drugs, which can be directly applied to clinical practice. Jintiange capsule is composed of multiple animal bones, and the characteristics of multiple components and multiple targets make it not found serious safety problems in clinical application. In addition, the elderly have good compliance by taking Jintiange capsule, and long-term use of Jintiange capsule to delay skeletal muscle aging is feasible. BRIEF DESCRIPTION OF DRAWINGS

[0008] Figure 1 is the JTG improved the mouse SASP index after administration.

[0009] Figure 2 is the RT-PCR analysis and quantitative detection of IL-6, IL-1β, MCP-1, VEGF, MMP-9 and CTGF (n=3). Statistical data are expressed as mean ± standard error (SEM). Statistical data are expressed as mean ± standard error (SEM). Note: ###P<0.001 compared with the Young group; *P<0.05 compared with the Old group, **P<0.01 compared with the Old group, ***P<0.01 compared with the Old group.

[0010] Figure 3 is the JTG improved the mouse SASP index after administration.

[0011] Figure 4 is the RT-PCR analysis and quantitative detection of IL-6, IL-1β, MCP-1, VEGF, MMP-9 and CTGF (n=3). Statistical data are expressed as mean ± standard error (SEM). Statistical data are expressed as mean ± standard error (SEM). Note: ###P<0.001 compared with the Young group; *P<0.05 compared with the Old group, **P<0.01 compared with the Old group, ***P<0.01 compared with the Old group.

[0012] Figure 5 is the JTG improved the mouse SASP index after administration.

[0013] Figure 6 is the RT-PCR analysis and quantitative detection of IL-6, IL-1β, MCP-1, VEGF, MMP-9 and CTGF (n=3). Statistical data are expressed as mean ± standard error (SEM). Note: *P<0.05 compared with the L-Old group, **P<0.01 compared with the L-Old group, ***P<0.01 compared with the L-Old group.

[0014] Figure 7 is the JTG improved the mouse SASP index after administration.

[0015] Figure 8 Western blot analysis and quantification of β-gal, P53 and P16 protein levels (n=3). Statistical data are expressed as mean ± standard error (SEM). Note: *P<0.05 compared with L-Old group, **P<0.01 compared with L-Old group, ***P<0.01 compared with L-Old group. DETAILED DESCRIPTION

[0016] The animal research protocol was approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine. During the entire experimental process, all operations involving animals strictly followed the established guidelines for humane treatment and welfare. Animals were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. All animals were housed in the Experimental Animal Center of Shanghai University of Traditional Chinese Medicine in a SPF environment, with a temperature of (21℃±2℃), a humidity of (60%±10%), a 12-hour light-dark alternating illumination, and free access to food and sterile drinking water.

[0017] After the mice were fed to 12 months of age, the old mice (n=40) were divided into four groups according to the experimental conditions. They included an aging model group (Old), and three groups of mice receiving different doses of Jintiange capsules (JTG) (0.0273g / kg, 0.0546g / kg and 0.1092g / kg, named JTG-L, JTG-M and JTG-H, respectively). In addition, 5-month-old mice were used as a young control group (n=8, Young). All mice were orally administered with the corresponding dose of JTG or distilled water for 12 weeks. After the end of drug administration, the animals were euthanized with 5% isoflurane, and serum and muscle tissue were collected.

[0018] To observe the long-term effects of drug administration after drug withdrawal, the mice were fed to 12 months of age. After reaching 12 months of age, the old mice (n=30) were divided into three groups according to the experimental conditions. They included an aging model group (L-Old) and two groups of mice receiving different doses of JTG (0.0546g / kg and 0.1092g / kg, named L-JTG-M and L-JTG-H, respectively). All mice were orally administered with the corresponding dose of JTG or distilled water for 12 weeks. After the end of drug administration, drug intervention was stopped, and the mice continued to be fed with feed. When the mice reached 18 months of age, euthanasia was performed using 5% isoflurane.

[0019] Real-time polymerase chain reaction (RT-PCR) detection method: total mRNA was extracted from skeletal muscle tissue and reverse transcribed into cDNA. The expression levels of IL-6, IL-1β, MCP-1, VEGF, MMP-9, CTGF and β2M as an internal reference gene were evaluated by quantitative PCR.

[0020] Western-Blot experimental detection method: total protein was extracted and quantified from colon tissue using RIPA lysis buffer and BCA protein quantification kit. The samples were denatured at 95°C for 10 minutes. Subsequently, the denatured samples were transferred to PVDF membranes after electrophoresis. Incubated with β-galactosidase (β-gal), P53 and P16 primary antibodies, washed with TBST, and incubated with the corresponding HRP secondary antibody. Finally, the protein signal was detected using an imaging system (Tanon, 4600SF).

[0021] Figure 1 、 2 is a graph of the detection of senescence-associated secretory phenotype (SASP) indicators in 12-month-old mice taking JTG for 3 months. Real-time quantitative PCR (RT-PCR) was used to detect the gene expression of six SASP factors related to cytokines, chemokines, growth factors, and proteases. Compared with the Young group, the expression of IL-6, IL-1β, MCP-1, VEGF, MMP-9, and CTGF in the Old group was significantly increased. Compared with the Old group, the expression of IL-6, IL-1β, VEGF, and MMP-9 in all JTG dose groups was significantly reduced. In addition, JTG-M and JTG-H significantly reduced the expression of MCP-1 and CTGF.

[0022] Figure 3 、 4 is a graph of the detection of senescence markers β-galactosidase (β-gal), P53, and P16 in 12-month-old mice taking JTG for 3 months. The results show that compared with the Young group, the levels of β-gal, P53, and P16 in the Old group were significantly increased. In the JTG dose groups, β-gal and P53 were significantly reduced, while P21 was significantly reduced in the JTG-M and JTG-H groups. These results indicate that JTG delays the aging process of skeletal muscle cells and delays skeletal muscle aging.

[0023] Figure 5 、 6 is a graph of the detection of SASP indicators in 12-month-old mice taking JTG for 3 months and stopping for 3 months. In muscle aging indicators, the expression of SASP indicators in the L-JTG-M and L-JTG-H groups was also reduced.

[0024] Figure 7 、 8 is a graph of the detection of senescence markers in 12-month-old mice taking JTG for 3 months and stopping for 3 months. The expression of β-gal, P53, and P16 proteins was reduced in the L-JTG-M and L-JTG-H groups. In summary, after stopping JTG, it can still continue to delay the aging of mouse skeletal muscle.

[0025] References:

[0026] [1] Kim H J, Jung D W, Williams D R. Age Is Just a Number: Progress and Obstacles in the Discovery of New Candidate Drugs for Sarcopenia [J]. Cells, 2023, 12(22).

[0027] Progress and Obstacles in the Discovery of New Candidate Drugs for Sarcopenia [J]. Cells, 2023, 12(22).

[0028] [2] Sun J, Yang X G, Hu Y C. Efficacy of Jintiange Capsules in the Treatment of Osteoporosis: A Network Meta-analysis [J]. Orthopaedic surgery, 2019, 11(2): 176-86.

Claims

1. Use of Jintiange capsules for delaying skeletal muscle aging.

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