National Institute of Animal Science, RDA, Suwon, 441-706, Korea.
Cell Biol Int. 2013 Sep;37(9):1003-9. doi: 10.1002/cbin.10150. Epub 2013 Jul 22.
Intercellular signalling communication between adipose and muscle tissue has been investigated. To test the effect of muscle cells on adipogenic gene expression, we utilised an in vitro co-culture system, in which fat (3T3-L1) and muscle (L-6) cells were physically separated but chemically exposed each other via insert with 0.4 µm porous membrane. When 3T3-L1 and L-6 cells reached at 80 and 40% confluence, respectively in separate wells, L-6 cells grown in insert were transferred onto 6-well plates where 3T3-L1 cells were being grown. When both cells were fully differentiated in co-culture plates, morphology of 3T3-L1 was examined by staining with Oil-red-O. Activity of glycerol-3-phosphate dehydrogenase (GPDH) and adipogenic gene expression including lipoprotein lipase (LPL), adipsin, GPDH, peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer binding protein (C/EBPα) were analysed. The presence of muscle cells during preadipocyte differentiation inhibited (P < 0.05) lipogenesis by suppressing lipogenic gene expression including LPL, adipsin and GPDH. Furthermore, GPDH activity was also decreased (P < 0.05) in 3T3-L1 cells by the presence of L-6 cells. These results suggest that presence of muscle cells suppresses adipogenic differentiation by inhibiting the adipogenic gene expression and GPDH activity in the muscle and fat cell co-culture system.
已经研究了脂肪组织和肌肉组织之间的细胞间信号通讯。为了测试肌肉细胞对脂肪生成基因表达的影响,我们利用了一种体外共培养系统,其中脂肪(3T3-L1)和肌肉(L-6)细胞通过带有 0.4μm 多孔膜的插入物物理分离但化学上相互暴露。当 3T3-L1 和 L-6 细胞分别在单独的孔中达到 80%和 40%的汇合度时,在插入物中生长的 L-6 细胞被转移到 6 孔板上,其中正在生长 3T3-L1 细胞。当两种细胞在共培养板中完全分化时,通过用油红 O 染色检查 3T3-L1 的形态。分析甘油-3-磷酸脱氢酶(GPDH)的活性和包括脂蛋白脂肪酶(LPL)、脂联素、GPDH、过氧化物酶体增殖物激活受体-γ(PPARγ)和 CCAAT/增强子结合蛋白(C/EBPα)在内的脂肪生成基因表达。在前脂肪细胞分化过程中存在肌肉细胞通过抑制包括 LPL、脂联素和 GPDH 在内的脂肪生成基因表达来抑制脂肪生成(P<0.05)。此外,L-6 细胞的存在还降低了 3T3-L1 细胞中的 GPDH 活性(P<0.05)。这些结果表明,肌肉细胞的存在通过抑制肌肉和脂肪细胞共培养系统中的脂肪生成基因表达和 GPDH 活性来抑制脂肪生成分化。