Application of musclin in preparation of drugs for inhibiting muscle adiposis, fibrosis and atrophy

Abstract

The invention relates to the technical field of polypeptide drugs, in particular to application of musclin in preparation of drugs for inhibiting muscle adiposis, fibrosis and atrophy, and the amino acid sequence of the musclin is shown as SEQ ID NO.1. According to the scheme, it is found that musclin can inhibit muscle adiposis and fibrosis by regulating proliferation and apoptosis of fibroblast / adipogenic precursor cells, the action target of Musclin is expanded, and then the effect of preventing and treating muscular atrophy can be achieved. The Musclin can be used as a polypeptide drug to be applied to the medical practice of resisting muscle atrophy, and can further be independently applied to the practice of muscle tissue repair of resisting muscle adiposis or fibrosis.

Description

technical field

[0001] The invention relates to the technical field of polypeptide drugs, in particular to the application of Musclin in the preparation of drugs for inhibiting muscle fat, fibrosis and atrophy. Background technique

[0002] Muscular atrophy can be caused by a variety of reasons, including heredity, denervation, senility, trauma, disuse, etc. Its main pathological feature is the decrease in the number of muscle fibers and the decrease in the volume of muscle fibers, thereby forming muscle atrophy. A state of decreased muscle mass, usually accompanied by decreased muscle strength. At the same time, a large number of fat cell infiltration and fiber deposition will appear in the atrophic muscle, which will hinder muscle regeneration and further promote the progression of muscle atrophy. Usually, fat cell infiltration and fiber deposition are chronic progressive development, and there is no effective way to cure them, leading to Muscular atrophy is difficult to ...

Examples

Embodiment 1

[0024] Example 1: Research on the effect of Musclin on inhibiting muscle fat formation

[0025] Musclin used in this program was purchased from Shanghai GL Biochem Co., Ltd, sequence: NH 2 - SFSGFGSPLDRLSAGSVEHRGKQRKAVDHSKKR-COOH (SEQ ID NO.1), which is the partial sequence of amino acids 80-112.

[0026] Musclin overexpression mice were constructed: Musclin recombinant protein was dissolved in 1% BSA solution at 80 μg / mL, Musclin solution was injected continuously for 5 days, once a day, 100 μL each time, and the control group was injected with the same amount of 1% BSA solution. ELISA experiments confirmed that compared with the control group, the Musclin concentration in the mouse muscles was significantly increased (experimental results are as follows: figure 1 As shown, the right is the group injected with Musclin protein, and the left is the group injected with 1% BSA solution), which proves that this method can effectively promote the increase of Musclin protein concen...

Embodiment 2

[0031] Example 2: Research on the effect of Musclin on inhibiting muscle fibrosis

[0032] The process of constructing Musclin overexpression mice is the same as that in Example 1.

[0033] Construction of muscle fibrosis mouse model: 50 μL of cardiotoxin (cardiotoxin, CTX) solution (10 μ M) was injected into the tibialis anterior muscle of mice (Musclin overexpression mice or control groups injected with 1% BSA) to induce muscle fibrosis.

[0034] Preparation of specimens: (1) take the tibialis anterior muscle on the 15th day after cardiotoxin injection, fix it in 4% paraformaldehyde for 12 hours, and then dehydrate it overnight with 30% sucrose solution; (2) take the tibialis anterior muscle after dehydration Tissues were prepared into frozen sections with a thickness of 8-10 μm using a cryostat.

[0035] Fluorescent immunohistochemical detection of fibrosis: (1) warm the frozen section to room temperature; (2) rehydrate in PBS; (3) block the sample with blocking solution a...

Embodiment 3

[0037] Embodiment 3: Musclin's research on the effect of treating muscular atrophy

[0038] Construction of muscle atrophy model: After mice were anesthetized with 0.5% pentobarbital sodium, the sciatic nerve and tibialis anterior tendon of one hind limb were cut off, and the sutures were sterilized.

[0039] Musclin treatment: Since the establishment of the model, the Musclin recombinant protein (80 μg / mL) was injected intraperitoneally every other day, and each injection was 100 μL until the time point of sample collection (to 42 days).

[0040] Preparation of specimens: (1) Take the atrophic tibialis anterior muscle at the specified time point, fix it in 4% paraformaldehyde for 12 hours, and then dehydrate it overnight with 30% sucrose solution; (2) press the dehydrated tibialis anterior muscle tissue by 8- Frozen sections were prepared with a thickness of 10 μm using a cryostat.

[0041] Fluorescent immunohistochemical detection of degeneration of atrophic muscle: (1) war...

technical field

[0001] The invention relates to the technical field of polypeptide drugs, in particular to the application of Musclin in the preparation of drugs for inhibiting muscle fat, fibrosis and atrophy. Background technique

[0002] Muscular atrophy can be caused by a variety of reasons, including heredity, denervation, senility, trauma, disuse, etc. Its main pathological feature is the decrease in the number of muscle fibers and the decrease in the volume of muscle fibers, thereby forming muscle atrophy. A state of decreased muscle mass, usually accompanied by decreased muscle strength. At the same time, a large number of fat cell infiltration and fiber deposition will appear in the atrophic muscle, which will hinder muscle regeneration and further promote the progression of muscle atrophy. Usually, fat cell infiltration and fiber deposition are chronic progressive development, and there is no effective way to cure them, leading to Muscular atrophy is difficult to ...

Examples

Embodiment 1

[0024] Example 1: Research on the effect of Musclin on inhibiting muscle fat formation

[0025] Musclin used in this program was purchased from Shanghai GL Biochem Co., Ltd, sequence: NH 2 - SFSGFGSPLDRLSAGSVEHRGKQRKAVDHSKKR-COOH (SEQ ID NO.1), which is the partial sequence of amino acids 80-112.

[0026] Musclin overexpression mice were constructed: Musclin recombinant protein was dissolved in 1% BSA solution at 80 μg / mL, Musclin solution was injected continuously for 5 days, once a day, 100 μL each time, and the control group was injected with the same amount of 1% BSA solution. ELISA experiments confirmed that compared with the control group, the Musclin concentration in the mouse muscles was significantly increased (experimental results are as follows: figure 1 As shown, the right is the group injected with Musclin protein, and the left is the group injected with 1% BSA solution), which proves that this method can effectively promote the increase of Musclin protein concen...

Embodiment 2

[0031] Example 2: Research on the effect of Musclin on inhibiting muscle fibrosis

[0032] The process of constructing Musclin overexpression mice is the same as that in Example 1.

[0033] Construction of muscle fibrosis mouse model: 50 μL of cardiotoxin (cardiotoxin, CTX) solution (10 μ M) was injected into the tibialis anterior muscle of mice (Musclin overexpression mice or control groups injected with 1% BSA) to induce muscle fibrosis.

[0034] Preparation of specimens: (1) take the tibialis anterior muscle on the 15th day after cardiotoxin injection, fix it in 4% paraformaldehyde for 12 hours, and then dehydrate it overnight with 30% sucrose solution; (2) take the tibialis anterior muscle after dehydration Tissues were prepared into frozen sections with a thickness of 8-10 μm using a cryostat.

[0035] Fluorescent immunohistochemical detection of fibrosis: (1) warm the frozen section to room temperature; (2) rehydrate in PBS; (3) block the sample with blocking solution a...

Embodiment 3

[0037] Embodiment 3: Musclin's research on the effect of treating muscular atrophy

[0038] Construction of muscle atrophy model: After mice were anesthetized with 0.5% pentobarbital sodium, the sciatic nerve and tibialis anterior tendon of one hind limb were cut off, and the sutures were sterilized.

[0039] Musclin treatment: Since the establishment of the model, the Musclin recombinant protein (80 μg / mL) was injected intraperitoneally every other day, and each injection was 100 μL until the time point of sample collection (to 42 days).

[0040] Preparation of specimens: (1) Take the atrophic tibialis anterior muscle at the specified time point, fix it in 4% paraformaldehyde for 12 hours, and then dehydrate it overnight with 30% sucrose solution; (2) press the dehydrated tibialis anterior muscle tissue by 8- Frozen sections were prepared with a thickness of 10 μm using a cryostat.

[0041] Fluorescent immunohistochemical detection of degeneration of atrophic muscle: (1) war...

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