McIntosh M, Hausman D, Martin R, Hausman G
Department of Food, Nutrition and Food Service Management, University of North Carolina at Greensboro, Greensboro, North Carolina 27402-6170, USA.
Am J Physiol. 1998 Aug;275(2):E285-93. doi: 10.1152/ajpendo.1998.275.2.E285.
The purpose of this study was to determine whether the antiobesity actions of dehydroepiandrosterone (DHEA) are due to an influence on preadipocyte proliferation and/or differentiation in primary cultures of pig and rat stromal-vascular (SV) cells. Pig SV cells were isolated from dorsal subcutaneous adipose tissue of 7-day-old pigs. For the proliferation assays, pig SV cells were grown for 4 days in plating medium containing DHEA at 0, 15, 50, or 150 microM. For the differentiation assays, pig SV cells were grown in plating medium for 3 days and then switched to a serum-free medium containing DHEA at 0, 15, 50, or 150 microM for the next 6 days. Rat SV cells were isolated from inguinal fat pads of 5-wk-old male rats. Rat SV cells were exposed to DHEA at 0, 5, 25, or 75 microM during proliferation. For the differentiation assays, rat SV cells were grown for 8 days in a serum-free medium containing DHEA at 0, 5, 25, or 75 microM. Preadipocyte differentiation [lipid staining, glycerol-3-phosphate dehydrogenase (GPDH) activity] and proliferation (preadipocyte-specific antigen staining) decreased with increasing levels of DHEA in cultures of pig SV cells. In cultures of rat SV cells, preadipocyte differentiation (lipid staining, GPDH activity) and proliferation ([3H]thymidine incorporation) were decreased in the 25 and 75 microM DHEA groups compared with the control and 5 microM DHEA groups. The level of expression of CCAAT enhancer binding protein-alpha, a master regulator of adipogenesis, in cultures of pig SV cells treated with 150 microM DHEA was 38% of control cultures. These data support the hypothesis that DHEA directly attenuates adipogenesis via attenuation of preadipocyte proliferation and differentiation.
本研究的目的是确定脱氢表雄酮（DHEA）的抗肥胖作用是否归因于对猪和大鼠基质血管（SV）细胞原代培养中前脂肪细胞增殖和/或分化的影响。猪SV细胞从7日龄仔猪的背部皮下脂肪组织中分离出来。在增殖试验中，猪SV细胞在含有0、15、50或150微摩尔/升DHEA的接种培养基中培养4天。在分化试验中，猪SV细胞在接种培养基中培养3天，然后在接下来的6天转换为含有0、15、50或150微摩尔/升DHEA的无血清培养基。大鼠SV细胞从5周龄雄性大鼠的腹股沟脂肪垫中分离出来。在增殖过程中，大鼠SV细胞暴露于0、5、25或75微摩尔/升的DHEA。在分化试验中，大鼠SV细胞在含有0、5、25或75微摩尔/升DHEA的无血清培养基中培养8天。在猪SV细胞培养物中，随着DHEA水平的升高，前脂肪细胞分化[脂质染色、甘油-3-磷酸脱氢酶（GPDH）活性]和增殖（前脂肪细胞特异性抗原染色）降低。在大鼠SV细胞培养物中，与对照和5微摩尔/升DHEA组相比，25和75微摩尔/升DHEA组中的前脂肪细胞分化（脂质染色、GPDH活性）和增殖（[3H]胸苷掺入）降低。在用150微摩尔/升DHEA处理的猪SV细胞培养物中，脂肪生成的主要调节因子CCAAT增强子结合蛋白-α的表达水平是对照培养物的38%。这些数据支持了DHEA通过减弱前脂肪细胞增殖和分化直接减弱脂肪生成的假说。
