脂肪酸对牛骨骼肌卫星细胞分化的影响
发布时间：2019-02-24 15:56
【摘要】：目的:近年来,脂肪酸对细胞分化的影响作用一直受到广泛关注。研究发现,脂肪酸可以通过调节过氧化物酶体增殖激活受体(PPAR)或其他转录因子而调控组织分化。单不饱和脂肪酸亚油酸(LA)和油酸(OA)在促进骨胳肌细胞的增殖和分化过程中具有重要作用。多不饱和脂肪酸如花生四烯酸(AA)和二十二碳六烯酸(DHA)还可以通过改变细胞膜的脂质分布,从而促进大鼠成肌细胞的分化。然而,脂肪酸是否能够影响牛骨骼肌细胞的发育,以及脂肪酸的摄取与骨骼肌卫星细胞分化的关系,仍不清楚。方法:本研究通过向牛骨骼肌卫星细胞(muscle-derived satellite cells,MDSCs)培养体系中添加三种不同的脂肪酸(棕榈酸PA、油酸OA和二十二碳六烯酸DHA),以探讨不同脂肪酸对牛骨骼肌卫星细胞分化的影响。结果:PA、OA和DHA作用浓度均为50 umol/L,以2%马血清为对照,诱导分化牛骨骼肌卫星细胞。分化96h后分别收集细胞,采用免疫荧光染色,实时定量PCR和Western blot技术检测细胞分化过程中相关的标志性基因MyoG(Myogenin)和MyH C(Myosin Heavy Chian)的表达变化进而探讨脂肪酸对体外培养的牛骨骼肌卫星细胞分化程度的影响;此外,通过荧光定量RT-PCR法检测脂肪酸代谢途径中关键基因CPT2(Carnitine palmitoyltransferase 2,肉碱棕榈酰转移酶Ⅱ)、DGAT2(Diacylglycerol acyltransferase 2,二酰甘油酰基转移酶Ⅱ)和ELOVL3(Fatty acid elongase 3,脂肪酸链延长酶III)的表达变化,进而探讨脂肪酸摄取对骨骼肌卫星细胞脂肪酸代谢的影响。实验结果显示,PA、OA和DHA处理的牛骨胳肌卫星细胞能够促进肌卫星细胞的分化。3种脂肪酸处理的细胞中脂肪酸代谢中的关键基因CPT2、DGAT2和ELOVL3的表达量显著增加。结论:脂肪酸(PA、OA、DHA)能够促进牛骨骼肌卫星细胞的分化,从而为探明肌肉发育和分化的分子机制提供帮助。
[Abstract]:Objective: in recent years, the effect of fatty acids on cell differentiation has been widely concerned. It has been found that fatty acids regulate tissue differentiation by regulating peroxisome proliferation-activated receptor (PPAR) or other transcription factors. Monounsaturated fatty acid linoleic acid (LA) and oleic acid (OA) play an important role in promoting the proliferation and differentiation of skeletal muscle cells. Polyunsaturated fatty acids such as arachidonic acid (AA) and 22 carbohexaenoic acid (DHA) can also promote the differentiation of rat myoblasts by changing the lipid distribution of cell membrane. However, it is not clear whether fatty acids can affect the development of bovine skeletal muscle cells, and the relationship between fatty acid uptake and skeletal muscle satellite cell differentiation. Methods: in this study, three different fatty acids (PA, oleic acid OA and 22 carbohexaenoic acid DHA),) were added to bovine skeletal muscle satellite cells (muscle-derived satellite cells,MDSCs) culture system. To investigate the effects of different fatty acids on the differentiation of bovine skeletal muscle satellite cells. Results: the concentration of PA,OA and DHA was 50 umol/L, and 2% horse serum was used as control to induce the differentiation of bovine skeletal muscle satellite cells. After 96 hours of differentiation, the cells were collected, and immunofluorescence staining was used. Real-time quantitative PCR and Western blot techniques were used to detect the expression of MyoG (Myogenin) and MyH C (Myosin Heavy Chian) related to cell differentiation and to explore the effect of fatty acids on the differentiation of bovine skeletal muscle satellite cells in vitro. In addition, CPT2 (Carnitine palmitoyltransferase _ 2, carnitine palmitoyltransferase 鈪，
本文编号：2429698
[Abstract]:Objective: in recent years, the effect of fatty acids on cell differentiation has been widely concerned. It has been found that fatty acids regulate tissue differentiation by regulating peroxisome proliferation-activated receptor (PPAR) or other transcription factors. Monounsaturated fatty acid linoleic acid (LA) and oleic acid (OA) play an important role in promoting the proliferation and differentiation of skeletal muscle cells. Polyunsaturated fatty acids such as arachidonic acid (AA) and 22 carbohexaenoic acid (DHA) can also promote the differentiation of rat myoblasts by changing the lipid distribution of cell membrane. However, it is not clear whether fatty acids can affect the development of bovine skeletal muscle cells, and the relationship between fatty acid uptake and skeletal muscle satellite cell differentiation. Methods: in this study, three different fatty acids (PA, oleic acid OA and 22 carbohexaenoic acid DHA),) were added to bovine skeletal muscle satellite cells (muscle-derived satellite cells,MDSCs) culture system. To investigate the effects of different fatty acids on the differentiation of bovine skeletal muscle satellite cells. Results: the concentration of PA,OA and DHA was 50 umol/L, and 2% horse serum was used as control to induce the differentiation of bovine skeletal muscle satellite cells. After 96 hours of differentiation, the cells were collected, and immunofluorescence staining was used. Real-time quantitative PCR and Western blot techniques were used to detect the expression of MyoG (Myogenin) and MyH C (Myosin Heavy Chian) related to cell differentiation and to explore the effect of fatty acids on the differentiation of bovine skeletal muscle satellite cells in vitro. In addition, CPT2 (Carnitine palmitoyltransferase _ 2, carnitine palmitoyltransferase 鈪，
本文编号：2429698
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