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牛的肌肉、皮下和肾周基质血管细胞表达相似的糖皮质激素受体亚型,但具有不同的成脂能力。

Bovine intramuscular, subcutaneous, and perirenal stromal-vascular cells express similar glucocorticoid receptor isoforms, but exhibit different adipogenic capacity.

作者信息

Ortiz-Colón G, Grant A C, Doumit M E, Buskirk D D

机构信息

Department of Animal Science, Michigan State University, East Lansing 48824-1225, USA.

出版信息

J Anim Sci. 2009 Jun;87(6):1913-20. doi: 10.2527/jas.2008-1350. Epub 2009 Feb 27.

Abstract

Understanding preadipocyte differentiation in economically important adipose depots will facilitate efforts to selectively increase intramuscular (i.m.) lipid accretion in cattle. The objectives of this study were to determine if glucocorticoid receptor (GR) expression differs among bovine stromal-vascular (S-V) cells derived from i.m., subcutaneous (s.c.), and peri-renal (p.r.) adipose tissue, and to evaluate the effects of dexamethasone (DEX) on adipogenesis of these cell populations. Stromal-vascular cells isolated from i.m., s.c., and p.r. adipose tissues of 2 steers were propagated in culture and exposed to 0 or 250 nM DEX for 48 h. Cell lysates were subjected to GR immunoblot analysis, and immunoreactive protein bands of approximately 97, approximately 62, and approximately 48 kDa were detected and expressed relative to beta-actin immunoreactivity. The abundance of each GR immunoreactive protein was similar among S-V cell populations (P > 0.50). Dexamethasone exposure decreased the abundance of the approximately 97 and approximately 62 kDa GR immunoreactive bands in S-V cells from the 3 depots (P < 0.001), but did not affect the expression of the approximately 48 kDa band (P = 0.96). Stromal-vascular cells isolated from 3 steers were grown in culture, and upon confluence, were exposed to 0, 25, or 2,500 nM DEX for 48 h. After an additional 10 d in differentiation media, differentiation was determined by glycerol-3-phosphate dehydrogenase (GPDH) specific activity and oil red O staining. The extent of differentiation differed by depot (p.r. > s.c. > i.m.; P < 0.05). Compared with control, 2,500 nM DEX increased GPDH activity in S-V cells from all depots (P < 0.05), and no interaction between depot and DEX concentration was observed (P = 0.99). We observed an adipose tissue depot by DEX concentration interaction (P = 0.03) for S-V cells with large (> or = 10 microm-diameter) lipid droplets. The percentage of p.r. S-V cells with large lipid droplets increased in response to DEX in a linear manner (P < 0.02), but only increased greater than control in s.c. cells exposed to 2,500 nM DEX (P = 0.002). Dexamethasone did not significantly increase the percentage of i.m. S-V cells with large lipid droplets (P > 0.27). Collectively, these data demonstrate differences in adipogenic activity among bovine i.m., s.c., and p.r. S-V cells, but indicate no relationship between adipogenic activity and glucocorticoid receptor abundance or function.

摘要

了解经济上重要的脂肪库中前脂肪细胞的分化,将有助于选择性增加牛肌肉内(i.m.)脂质积累的研究。本研究的目的是确定源自i.m.、皮下(s.c.)和肾周(p.r.)脂肪组织的牛基质血管(S-V)细胞中糖皮质激素受体(GR)的表达是否存在差异,并评估地塞米松(DEX)对这些细胞群体脂肪生成的影响。从2头公牛的i.m.、s.c.和p.r.脂肪组织中分离出的基质血管细胞在培养中进行增殖,并暴露于0或250 nM DEX中48小时。细胞裂解物进行GR免疫印迹分析,检测到约97 kDa、约62 kDa和约48 kDa的免疫反应性蛋白条带,并相对于β-肌动蛋白免疫反应性进行表达。各GR免疫反应性蛋白的丰度在S-V细胞群体之间相似(P>0.50)。DEX处理降低了来自3个脂肪库的S-V细胞中约97 kDa和约62 kDa GR免疫反应性条带的丰度(P<0.001),但不影响约48 kDa条带的表达(P = 0.96)。从3头公牛中分离出的基质血管细胞在培养中生长,汇合后暴露于0、25或2500 nM DEX中48小时。在分化培养基中再培养10天后,通过甘油-3-磷酸脱氢酶(GPDH)比活性和油红O染色来确定分化情况。分化程度因脂肪库而异(p.r.>s.c.>i.m.;P<0.05)。与对照相比,2500 nM DEX增加了所有脂肪库的S-V细胞中的GPDH活性(P<0.05),且未观察到脂肪库与DEX浓度之间的相互作用(P = 0.99)。对于具有大(直径≥10微米)脂滴的S-V细胞,我们观察到脂肪组织库与DEX浓度之间存在相互作用(P = 0.03)。p.r. S-V细胞中具有大脂滴的百分比随DEX呈线性增加(P<0.02),但仅在暴露于两千五百nM DEX的s.c.细胞中增加幅度大于对照(P = 0.002)。DEX并未显著增加i.m. S-V细胞中具有大脂滴的百分比(P>0.27)。总体而言,这些数据表明牛i.m.、s.c.和p.r. S-V细胞之间在脂肪生成活性上存在差异,但表明脂肪生成活性与糖皮质激素受体丰度或功能之间没有关系。

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