CAJ | 학술논문

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.
연구록화고체외모의저양대소서3T3-L1지방세포단핵세포추화단백1표체적영향.체외배양3T3-L1전지방세포,병장기유도분화위성숙지방세포,유홍O염색감정지방세포분화정도화지질적취정황.록화고화학모의지방세포저양배경,채용실시형광정량PCR여단백면역인적법검측저양유도인자-1α적mRNA화단백수평;채용실시형광정량PCR여매련면역흡부법검측단핵세포추화단백1적mRNA화단백수평,병대량자적단백수평진행상관성분석.록화고처리분화성숙적3T3-L1지방세포후,저양유도인자-1α재mRNA급단백수평균현저상조;동시발현단핵세포추화단백1 mRNA적표체수평화배양액중단핵세포추화단백1단백적분비수평균승고,차저양유도인자-1α여단핵세포추화단백1단백수평정현선성상관( r=0.864,P＜0.01).록화고유도저양상조3T3-L1지방세포단핵세포추화단백1적표체,삼여지방조직만성저도염증적발생.
To investigate the effects of hypoxia chemically induced by CoCl2 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.CoCl2 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.CoCl2 induced hypoxia led to a marked recruitment of HIF-1α in mouse 3T3-L1adipocytes.Similarly,both mRNA and protein levels of MCP-1 were up-regulated.Exposure of 3T3-L1 adipocytes to CoCl2 induced hypoxic microenvironment in vitro,and hypoxic induction of MCP-1 expression and secretion may be mediated by HIF-1 α and may contribute to chronic low grade inflammation in adipose tissue.