Choi S H, Park S K, Johnson B J, Chung K Y, Choi C W, Kim K H, Kim W Y, Smith B
Department of Animal Science, Texas Agricultural Experiment Station, Texas A&M University, College Station, TX 77843, USA ; Department of Animal Science, Chungbuk National University, Cheongju 361-763, Korea .
Department of Food Science and Technology, Sejong University, Seoul, 143-747, Korea .
Asian-Australas J Anim Sci. 2015 Mar;28(3):411-9. doi: 10.5713/ajas.14.0598.
We previously demonstrated that bovine subcutaneous preadipocytes promote adipogenic gene expression in muscle satellite cells in a co-culture system. Herein we hypothesize that saturated fatty acids would promote adipogenic/lipogenic gene expression, whereas mono- and polyunsaturated fatty acids would have the opposite effect. Bovine semimembranosus satellite cells (BSC) and intramuscular preadipocytes (IPA) were isolated from crossbred steers and cultured with 10% fetal bovine serum (FBS)/Dulbecco's Modified Eagle Medium (DMEM) and 1% antibiotics during the 3-d proliferation period. After proliferation, cells were treated for 3 d with 3% horse serum/DMEM (BSC) or 5% FBS/DMEM (IPA) with antibiotics. Media also contained 10 μg/mL insulin and 10 μg/mL pioglitazone. Subsequently, differentiating BSC and IPA were cultured in their respective media with 40 μM palmitic, stearic, oleic, or linoleic acid for 4 d. Finally, BSC and IPA were single- or co-cultured for an additional 2 h. All fatty acid treatments increased (p = 0.001) carnitine palmitoyltransferase-1 beta (CPT1β) gene expression, but the increase in CPT1β gene expression was especially pronounced in IPA incubated with palmitic and stearic acid (6- to 17- fold increases). Oleic and linoleic acid decreased (p = 0.001) stearoyl-CoA desaturase (SCD) gene expression over 80% in both BSC and IPA. Conversely, palmitic and stearic acid increased SCD gene expression three fold in co-cultured in IPA, and stearic acid increased AMPKα gene expression in single- and co-cultured BSC and IPA. Consistent with our hypothesis, saturated fatty acids, especially stearic acid, promoted adipogenic and lipogenic gene expression, whereas unsaturated fatty acids decreased expression of those genes associated with fatty acid metabolism.
我们之前证明,在共培养体系中,牛皮下前脂肪细胞可促进肌肉卫星细胞中脂肪生成基因的表达。在此,我们假设饱和脂肪酸会促进脂肪生成/脂质生成基因的表达,而单不饱和脂肪酸和多不饱和脂肪酸则会产生相反的效果。从杂交阉牛中分离出牛半膜肌卫星细胞(BSC)和肌内前脂肪细胞(IPA),并在3天的增殖期内用10%胎牛血清(FBS)/杜氏改良 Eagle 培养基(DMEM)和1%抗生素进行培养。增殖后,细胞用含抗生素的3%马血清/DMEM(BSC)或5% FBS/DMEM(IPA)处理3天。培养基中还含有10μg/mL胰岛素和10μg/mL吡格列酮。随后,将分化中的BSC和IPA在各自的培养基中与40μM棕榈酸、硬脂酸、油酸或亚油酸一起培养4天。最后,将BSC和IPA单独或共培养2小时。所有脂肪酸处理均增加了(p = 0.001)肉碱棕榈酰转移酶-1β(CPT1β)基因的表达,但在用棕榈酸和硬脂酸孵育的IPA中,CPT1β基因表达的增加尤为明显(增加了6至17倍)。油酸和亚油酸在BSC和IPA中均使硬脂酰辅酶A去饱和酶(SCD)基因表达降低了80%以上(p = 0.001)。相反,棕榈酸和硬脂酸使共培养的IPA中的SCD基因表达增加了三倍,硬脂酸使单独培养和共培养的BSC和IPA中的AMPKα基因表达增加。与我们的假设一致,饱和脂肪酸,尤其是硬脂酸,促进了脂肪生成和脂质生成基因的表达,而不饱和脂肪酸则降低了与脂肪酸代谢相关的那些基因的表达。