摘要
研究氯化钴体外模拟低氧对小鼠3T3-L1脂肪细胞单核细胞趋化蛋白1表达的影响。体外培养3T3-L1前脂肪细胞，并将其诱导分化为成熟脂肪细胞，油红O染色鉴定脂肪细胞分化程度和脂质积聚情况。氯化钴化学模拟脂肪细胞低氧环境，采用实时荧光定量PCR与蛋白免疫印迹法检测低氧诱导因子-1α的mRNA和蛋白水平；采用实时荧光定量PCR与酶联免疫吸附法检测单核细胞趋化蛋白1的mRNA和蛋白水平，并对两者的蛋白水平进行相关性分析。氯化钴处理分化成熟的3T3-L1脂肪细胞后，低氧诱导因子-1α在mRNA及蛋白水平均显著上调；同时发现单核细胞趋化蛋白1 mRNA的表达水平和培养液中单核细胞趋化蛋白1蛋白的分泌水平均升高，且低氧诱导因子-1α与单核细胞趋化蛋白1蛋白水平呈现线性相关（r=0．864，P〈0．01）。氯化钴诱导低氧上调3T3-L1脂肪细胞单核细胞趋化蛋白1的表达，参与脂肪组织慢性低度炎症的发生。
To investigate the effects of hypoxia chemically induced by COCl_2 on the expression of monocyte chemoattractant protein-1 （ MCP-1 ） in mouse 3T3-L1 adipocytes. 3T3-L1 preadipocytes were cultured in vitro and differentiated into the matured adipocytes. Cell differentiation and lipid accumulation was determined by Oil Red O staining. COCl_2 was used as a chemical hypoxia-inducible reagent to mimic hypoxic microenvironment. The effect of COCl2 on cell viability was estimated by MTT assay. Hypoxia-inducible factor-1α （ HIF-1α ） expression under hypoxia was detected by realtime fluorescent PCR and Western blot, while MCP-1 expression was detected by real-time fluorescent PCR and ELISA. CoCI： induced hypoxia led to a marked recruitment of HIF-lα in mouse 3T3-L1 adipocytes. Similarly, both mRNA and protein levels of MCP-1 were up-regulated. Exposure of 3T3-L1 adipocytes to COCl_2 induced hypoxic microenvironment in vitro, and hypoxic induction of MCP-1 expression and secretion may be mediated by HIF-lα and may contribute to chronic low zrade inflammation in adioose tissue.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2011年第10期853-856,共4页
Chinese Journal of Endocrinology and Metabolism
