Novel application of DHA as feed additive in regulation and control of muscle fiber types
A new use, muscle fiber technology, applied in the direction of animal feed, application, additional food elements, etc., can solve the problems of no relevant technical reports, research is still blank, etc., to increase the number of mitochondria, broaden the scope of application, and improve the quality of skeletal muscle Effect
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Embodiment 1
[0034] Example 1: Daily gavage of 500 mg / kg body weight of docosahexaenoic acid to mice can increase the exercise tolerance of mice.
[0035] 1. In order to study the effect of docosahexaenoic acid on the exercise endurance of mouse skeletal muscle, 8-week-old male C57BL / 6 mice were used as the research object.
[0036] 2. Experimental method
[0037] (1) Animal feeding method: 2 animals / cage; feeding temperature and humidity: temperature is 24±1°C, humidity is 40% to 70%; 12h: 12h day and night lighting is used; feeding conditions are always kept stable to ensure the test results reliability. During the breeding period, the animals had free access to drinking water, maintenance feed (NFD) and high-fat feed (HFD) were provided by Jiangsu Synergy Biological Co., Ltd. The purchased SPF grade C57BL / 6 mice were pre-fed for 1 week to adapt to the environment.
[0038]2) Animal grouping and treatment: 8-week-old mice were randomly divided into three groups: normal diet group (NFD...
Embodiment 2
[0049] Embodiment 2: the DHA of mouse skeletal muscle mRNA mRNA m 6 Effect of A modification.
[0050] 1. In order to study the effect of DHA on the muscle fiber types of mouse skeletal muscle, 8-week-old male C57BL / 6 mice were used as the research object.
[0051] 2. Experimental method
[0052] (1) Animal breeding method: the same as in Example 1.
[0053] (2) Grouping and treatment of animals: same as in Example 1.
[0054] (3) Sampling and detection: On the day of the end of the test in step (2), mice were humanely sacrificed by cervical dislocation, gastrocnemius muscles of all mice were collected, and total mRNA was extracted for mass spectrometry detection of mRNA m 6 A level, extract protein for Western Blot detection m 6 A-related protein expression.
[0055] 3. The result is as follows Figure 5-6 Shown:
[0056] Figure 5 is the effect of DHA administration on mouse skeletal muscle m 6 Effect of A modification level.
[0057] The result is as Figure 5 As...
Embodiment 3
[0060] Example 3: Whether DHA is verified by mRNA m at the myoblast level 6 A modification regulates myofiber type switching and mitochondrial generation.
[0061] 1. In order to study the mechanism of DHA on the transformation of mouse skeletal muscle fiber types, the C2C12 cell line was used as the research object.
[0062] 2. Experimental method
[0063] (1) Cell culture and induced differentiation: C2C12 myoblasts were amplified by high-glucose DMEM complete medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin solution. When the cell density reached 90%, the culture was replaced. The base was high-glucose DMEM induction medium supplemented with 2% horse-cow serum and 1% penicillin-streptomycin solution to induce differentiation, and the amount of DHA added was 5-10 μM.
[0064] (2) Cell transfection: use the liposome transfection method to introduce the synthetic siFto small molecule interference sequence into myoblasts.
[0065] (3) Mitochondr...
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