Oligonucleotide medicine for treating myophagism
An oligonucleotide and muscle atrophy technology, applied in the field of compositions containing the oligonucleotide, can solve the problems that there is still no effective method for treating muscle atrophy or enhancing muscle growth
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Embodiment 1
[0165] Example 1 Inhibition of Myostatin at the Cell Level
[0166] 1. The purpose of the experiment
[0167] This experiment is to screen active RNA oligonucleotides in mouse muscle cell lines. The detection index is to observe the effect of RNA oligonucleotide on the expression of myostatin by Western blotting.
[0168] 2. Experimental materials
[0169] 1) Cells: The C2C12 cell line is derived from mouse muscle cells and is cultured in DMEM medium. For every 100 ml of DMEM complete medium, 15 ml of fetal bovine serum, 100 units of penicillin, and 100 micrograms of streptomycin are added. The culture condition is 37℃, 5% CO 2 . Cell differentiation culture uses 2% horse serum medium, the culture condition is 37℃, 5% CO 2 .
[0170] 2) RNA oligonucleotides: seven RNA oligonucleotides that target different regions of the myostatin gene, each with a 3'end butane protecting group. RNA oligonucleotides used for cell transfection efficiency experiments are labeled with fluorescent gro...
Embodiment 2
[0188] Example 2 Comparison of inhibitory effects of single and combined RNA oligonucleotides on mouse muscle growth
[0189] 1. Experimental purpose: to study the effect of RNA oligonucleotides on muscle growth in vivo under single or combined use conditions.
[0190] 2. Experimental materials: three RNA oligonucleotides respectively target the 5'untranslated region (A) of the myostatin gene; translation initiation region (B); translation termination region (C). Each RNA oligonucleotide was dissolved in physiological saline at a concentration of 1 mg / ml. Mix each RNA oligonucleotide in an equal volume to make a combined RNA oligonucleotide.
[0191] 3. Experimental procedure: The experiment is divided into five groups, each group contains six 6-8 week old female mice. The experiment used tail vein injection every three days, and the injection volume was 100μl (5mg / kg body weight). After the experiment is completed, the leg muscles are stripped and weighed.
[0192] 4. Experimenta...
Embodiment 3
[0202] Example 3 Effect of RNA oligonucleotide composition on mouse muscle growth
[0203] 1. The purpose of the experiment
[0204] The three selected RNA oligonucleotides A, B, and C were used to further explore the effects of the combined use of the three oligonucleotides on mouse muscle growth and the expression of myostatin gene in muscle tissue.
[0205] 2. Experimental materials
[0206] 1) Experimental animals: A total of 24 mice, female mice aged 6-8 weeks.
[0207] 2) Oligonucleotides: Dissolve each oligonucleotide. Three kinds of RNA oligonucleotides (targeting the 5'untranslated region, translation initiation region, and translation termination region of myostatin respectively) are mixed in equal volume, and the injection volume is 5mg / kg, 100μl per mouse; four kinds of DNA Oligonucleotides (used as a control) are mixed in equal volume, and the injection volume is 5 mg / kg, 100 μl per mouse;
[0208] 3. Experimental methods and procedures
[0209] The experiment was div...
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