促酰化蛋白对3T3-L1脂肪细胞炎症反应的影响

发布时间：2018-03-13 01:08

本文选题：脂肪细胞　切入点：胰岛素抵抗　出处：《华中科技大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的观察不同种类、不同浓度的脂肪酸对3T3-L1成熟脂肪细胞胰岛素刺激状态和ASP刺激状态下细胞炎症因子(IL-6、TNF-α、MCP-1、MIP-1α)分泌水平的影响,以及对细胞炎症信号因子(JNK1、IKKβ)蛋白表达的影响,以探讨ASP对脂肪细胞炎症反应的调节作用。方法 1、采用经典的激素“鸡尾酒”法(诱导培养基由10μg/ml胰岛素、0.5 mmol/L 1-甲基3-异丁基黄嘌呤和1.0μmol/L地塞米松配置而成),将3T3-L1前脂肪细胞诱导分化为3T3-L1成熟脂肪细胞,倒置显微镜下观察3T3-L1前脂肪细胞的形态学变化,油红O染色观察脂滴的聚集。 2、3T3-L1成熟脂肪细胞给予油酸、棕榈酸孵育过夜后,采用ELISE方法检测ASP刺激以及胰岛素刺激的IL-6、TNF-α、MCP-1、MIP-1α分泌水平。 3、3T3-L1成熟脂肪细胞给予油酸、棕榈酸孵育过夜后,采用Western Blot法检测ASP刺激以及胰岛素刺激的JNK1、IKKβ蛋白表达。结果 1、3T3-L1前脂肪细胞表现为成纤维细胞样的长梭形,胞浆中无脂滴;分化成熟后,细胞变圆、变大,可见明显的脂滴围绕胞核,呈典型“戒环”形状。 2、在脂肪酸诱导的胰岛素抵抗状态下,ASP可以从不同程度上抑制IL-6、TNF-α、MCP-1、MIP-1α分泌。油酸组显著下调57% MIP-1α的分泌(P0.05,1.0 mmol/L ),棕榈酸组显著抑制50%IL-6的分泌(P0.05,1.0 mmol/L )和48% TNF-α的分泌(P0.05,1.0 mmol/L )。高浓度的油酸和棕榈酸可以明显抑制ASP刺激的JNK1蛋白表达,轻度抑制IKKβ的表达。油酸组JNK1的表达下降34%(P0.05,1.0 mmol/L),棕榈酸组下降54%(P0.01,1.0 mmol/L)。 3、在脂肪酸诱导的胰岛素抵抗状态下,胰岛素刺激的3T3-L1脂肪细胞IL-6、TNF-α、MCP-1的分泌随FFA浓度的升高呈下降趋势,油酸组显著增强MIP-1α的分泌178% (P0.05,1.0 mmol/L ),棕榈酸组显著抑制37% MCP-1(P0.05,1.0 mmol/L )和51% TNF-α分泌(P0.05,1.0 mmol/L )。与ASP刺激不同的是,胰岛素刺激状态下,油酸、棕榈酸不影响JNK1和IKKβ的表达。结论在脂肪酸诱导的脂肪细胞胰岛素抵抗状态下,ASP在一定程度上抑制炎症因子分泌,而且还参与调节JNK、IKK/NF-κB信号通路中重要信号分子的功能, ASP参与了脂肪细胞脂毒性-炎症反应的调控。
[Abstract]:Purpose. To observe the effects of different kinds of fatty acids on the secretion of IL-6 TNF- 伪 MCP-1mMIP-1 伪 in 3T3-L1 mature adipocytes stimulated by insulin and ASP, and on the expression of JNK1I-IKK 尾. To investigate the effect of ASP on the inflammatory response of adipocytes. Method. 1. The 3T3-L1 preadipocytes were induced to differentiate into 3T3-L1 mature adipocytes by the classical hormone "cocktail" method (the induction medium consisted of 10 渭 g / ml insulin and 0.5 mmol/L 1-methyl-isobutyl xanthine and 1.0 渭 mol/L dexamethasone). The morphological changes of 3T3-L1 preadipocytes were observed under inverted microscope, and the aggregation of lipid droplets was observed by oil red O staining. 2the mature adipocytes were treated with oleic acid and palmitic acid for the night. The levels of MIP-1 伪 secreted by ASP and insulin stimulated IL-6TNF- 伪 MCP-1were detected by ELISE. 3T3-L1 mature adipocytes were treated with oleic acid and palmitic acid for the night. Western Blot assay was used to detect the expression of ASP stimulated and insulin-stimulated JNK1KK 尾 protein. Results. 1the preadipocytes of 3T3-L1 showed long fusiform like fibroblasts with no lipid droplets in the cytoplasm, and after differentiation and maturation, the cells became round and large, and obvious lipid droplets surrounded the nucleus, taking the shape of typical "ring ring". 2. Under the condition of insulin resistance induced by fatty acid, ASP inhibited the secretion of IL-6 TNF- 伪 and MCP-1 / MIP-1 伪 in varying degrees. In oleic acid group, the secretion of 57% MIP-1 伪 was significantly down-regulated (P0.051.0 mmol/L), and palmitic acid group significantly inhibited the secretion of 50 TNF- 伪 (P0.051.0 mmol/L) and 48% TNF- 伪 (P0.051-1.0 mmol/L). High concentrations of oleic acid and palmitic acid could significantly inhibit the expression of JNK1 protein stimulated by ASP. The expression of IKK 尾 was slightly inhibited in oleic acid group (P 0.05) and palmitic acid group (P 0.01) 1.0 mmol / L 路L ~ (-1) 路L ~ (-1) 路min ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1). 3. Under the condition of insulin resistance induced by fatty acid, the secretion of IL-6TNF- 伪 MCP-1 in 3T3-L1 adipocytes induced by insulin decreased with the increase of FFA concentration. In oleic acid group, the secretion of MIP-1 伪 was significantly increased by 178% ~ (178%) (P _ (0.05) ~ (1.0) mmol/L), while in palmitic acid group it was significantly inhibited by 37% MCP-1 (P _ (0.05N) ~ (1.0) mmol/L) and 51% ~ (TNF- 伪) (P _ (0.05N) ~ (1.0) mmol/L). In contrast to ASP stimulation, oleic acid and palmitic acid did not affect the expression of JNK1 and IKK 尾. Conclusion. Under the condition of insulin resistance induced by fatty acids, ASP inhibited the secretion of inflammatory factors to some extent. ASP is involved in the regulation of lipid toxicity and inflammation in adipocytes.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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