【摘要】：目的:近年來,脂肪酸對細胞分化的影響作用一直受到廣泛關注。研究發(fā)現(xiàn),脂肪酸可以通過調節(jié)過氧化物酶體增殖激活受體(PPAR)或其他轉錄因子而調控組織分化。單不飽和脂肪酸亞油酸(LA)和油酸(OA)在促進骨胳肌細胞的增殖和分化過程中具有重要作用。多不飽和脂肪酸如花生四烯酸(AA)和二十二碳六烯酸(DHA)還可以通過改變細胞膜的脂質分布,從而促進大鼠成肌細胞的分化。然而,脂肪酸是否能夠影響牛骨骼肌細胞的發(fā)育,以及脂肪酸的攝取與骨骼肌衛(wèi)星細胞分化的關系,仍不清楚。方法:本研究通過向牛骨骼肌衛(wèi)星細胞(muscle-derived satellite cells,MDSCs)培養(yǎng)體系中添加三種不同的脂肪酸(棕櫚酸PA、油酸OA和二十二碳六烯酸DHA),以探討不同脂肪酸對牛骨骼肌衛(wèi)星細胞分化的影響。結果:PA、OA和DHA作用濃度均為50 umol/L,以2%馬血清為對照,誘導分化牛骨骼肌衛(wèi)星細胞。分化96h后分別收集細胞,采用免疫熒光染色,實時定量PCR和Western blot技術檢測細胞分化過程中相關的標志性基因MyoG(Myogenin)和MyH C(Myosin Heavy Chian)的表達變化進而探討脂肪酸對體外培養(yǎng)的牛骨骼肌衛(wèi)星細胞分化程度的影響;此外,通過熒光定量RT-PCR法檢測脂肪酸代謝途徑中關鍵基因CPT2(Carnitine palmitoyltransferase 2,肉堿棕櫚酰轉移酶Ⅱ)、DGAT2(Diacylglycerol acyltransferase 2,二酰甘油�；D移酶Ⅱ)和ELOVL3(Fatty acid elongase 3,脂肪酸鏈延長酶III)的表達變化,進而探討脂肪酸攝取對骨骼肌衛(wèi)星細胞脂肪酸代謝的影響。實驗結果顯示,PA、OA和DHA處理的牛骨胳肌衛(wèi)星細胞能夠促進肌衛(wèi)星細胞的分化。3種脂肪酸處理的細胞中脂肪酸代謝中的關鍵基因CPT2、DGAT2和ELOVL3的表達量顯著增加。結論:脂肪酸(PA、OA、DHA)能夠促進牛骨骼肌衛(wèi)星細胞的分化,從而為探明肌肉發(fā)育和分化的分子機制提供幫助。
[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，

本文編號：2429698