防己黄芪汤对小鼠3T3-L1前脂肪细胞分化及炎症因子基因表达的影响

发布时间：2018-03-20 06:38

本文选题：防己黄芪汤　切入点：3T3-L1前脂肪细胞　出处：《宁夏医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1.观察防己黄芪汤对小鼠3T3-L1前脂肪细胞分化的影响;2.观察防己黄芪汤对小鼠3T3-L1前脂肪细胞分化过程中炎症因子MCP-1、IL-6、TNF-αmRNA表达的影响。方法:体外培养3T3-L1前脂肪细胞,在细胞融合后2天,用传统“鸡尾酒”法(即含胰岛素、地塞米松、异丁基-3-甲基黄嘌呤的混合诱导剂)诱导3T3-L1前脂肪细胞分化为脂肪细胞。在诱导分化的第0天,设置溶媒对照组(1 mol/L PBS)和药物处理组(100μg/ml防己黄芪汤)。1.在成功诱导分化的第10天,用油红O染色,拍照观察,用异丙醇溶解油红O,使用酶标仪进行定量检测各组A492nm以间接反映3T3-L1前脂肪细胞分化程度及细胞内甘油三酯小脂滴含量。2.在成功诱导分化的第10天,提取细胞总RNA,采用real-time PCR法检测3T3-L1前脂肪细胞分化相关基因,过氧化物酶体增殖受体γ(PPARγ)、脂联素(adiponectin)mRNA的表达情况。3.在成功诱导分化的第10天,提取细胞总RNA,采用real-time PCR法检测3T3-L1脂肪细胞炎症因子,MCP-1、IL-6、TNF-αmRNA的表达情况。结果:1.油红O染色显示药物处理组(100μg/ml防己黄芪汤)中含有甘油三酯小脂滴的成熟脂肪细胞少于溶媒对照组(1 mol/L PBS),P0.05。2.在诱导分化的第10天,药物处理组(100μg/ml防己黄芪汤)的PPARγmRNA(P0.005)、脂联素mRNA(P0.005)明显低于溶媒对照组(1 mol/L PBS)。3.在诱导分化的第10天,药物处理组(100μg/ml防己黄芪汤)的MCP-1 mRNA(P0.05)、TNF-αmRNA(P0.005)明显低于溶媒对照组(1 mol/L PBS),IL-6 mRNA表达两组无统计学意义。结论:1.通过定性分析(油红O染色)、定量分析(3T3-L1前脂肪细胞分化相关基因PPARγ、脂联素mRNA表达)说明防己黄芪汤可有效抑制3T3-L1前脂肪细胞分化,减少脂肪细胞数量;2.防己黄芪汤可抑制3T3-L1脂肪细胞中炎症因子MCP-1、TNF-ɑm RNA的表达,具有改善3T3-L1脂肪细胞“微炎症状态”,可能对改善脂肪组织胰岛素抵抗具有积极作用。
[Abstract]:Objective 1. To observe the effect of Fangji Huangqi decoction on the differentiation of 3T3-L1 preadipocytes in mice 2.To observe the effect of Fangji Huangqi decoction on the expression of MCP-1- IL-6TNF- 伪 mRNA during the differentiation of 3T3-L1 preadipocytes in mice. Methods: 3T3-L1 preadipocytes were cultured in vitro. Two days after cell fusion, 3T3-L1 preadipocytes were induced to differentiate into adipocytes by traditional "cocktail" method (I. e., mixed inducers containing insulin, dexamethasone, isobutyl -3-methylxanthine). On day 0 of differentiation, 3T3-L1 preadipocytes were induced to differentiate into adipocytes. #number0# 渭 g / ml Fangji Astragalus decoction (100 渭 g / ml) was used in the control group and the drug treatment group. On the 10th day after successful differentiation induction, oil red O staining was used to take photos and observe the differentiation. Oil red O4 was dissolved with isopropanol, and A492nm was quantitatively detected by enzymatic analyzer to indirectly reflect the differentiation degree of 3T3-L1 preadipocytes and the content of triglyceride small lipid droplets in the cells. Cell total RNAs were extracted, and the expression of 3T3-L1 preadipocyte differentiation related genes, peroxisome proliferating receptor 纬 -PPAR- 纬, adiponectinine adiponectinine mRNA expression was detected by real-time PCR assay. Total cell RNAs were extracted, and the expression of 3T3-L1 inflammatory cytokine MCP-1and IL-6TNF- 伪 mRNA was detected by real-time PCR method. Results: 1. Oil red O staining showed that the mature adipocytes containing triglyceride small lipid drops in the drug treatment group were less than those in the drug treatment group (100 渭 g / ml Fangji astragalus decoction). In the solvent control group, P0. 05.2 of 1 mol/L PBSs was observed on the 10th day after induction of differentiation. The PPAR 纬 mRNAs (P0.005) and adiponectin mRNAs (P0.005) of 100 渭 g / ml Fangji Astragalus decoction in the drug treatment group were significantly lower than those in the solvent control group (1 渭 g / ml Fangji astragalus decoction). The expression of MCP-1 mRNA- 伪 mRNA-TNF- 伪 mRNA-P0.005 in the drug treatment group was significantly lower than that in the control group (P 0.005). Conclusion: 1: 1. Quantitative analysis of 3T3-L1 preadipocyte differentiation related units by qualitative analysis (oil red O staining). Because of the expression of PPAR 纬, adiponectin mRNA, Fangji Huangqi decoction could effectively inhibit the differentiation of 3T3-L1 preadipocytes. Fangji Huangqi decoction can inhibit the expression of MCP-1and TNF- RNA in 3T3-L1 adipocytes and improve the "microinflammatory state" of 3T3-L1 adipocytes, which may play a positive role in improving insulin resistance in adipose tissue.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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