邻苯二甲酸酯类对3T3-L1前体脂肪细胞诱导分化的影响

发布时间：2018-03-08 15:12

本文选题：邻苯二甲酸酯类　切入点：肥胖　出处：《重庆医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：研究目的：脂肪细胞的增殖和分化异常可引起脂肪细胞数目增多、体积变大、脂肪颗粒过多堆积，从而导致能量代谢平衡失调，引起肥胖及相关疾病，给社会经济发展带来巨大经济负担。环境因素是肥胖疾病的影响因素之一：邻苯二甲酸酯类(PAEs）是一类持久性有机污染物，具有环境类雌激素样作用，在人体内已有较高暴露量。目前关于其毒性危害研究均集中在生殖毒性方面。文献报道和前期研究提示PAEs与腹部肥胖以及儿童肥胖相关，但目前未见其影响机制的研究。研究方法：本课题以PAEs是过氧化物酶体增殖剂为切入点，可以激活过氧化物酶体增殖剂受体PPARs。以两种我国工业使用最多的、人体暴露量最高的两种PAEs（DBP、DEHP）为研究目标。采用体外培养小鼠来源的3T3-L1前体脂肪细胞建立实验模型，经典激素鸡尾酒法诱导3T3-L1前体脂肪细胞分化成成熟的脂肪细胞，设计两种标志性PAEs染毒参与3T3-L1前体脂肪细胞的诱导分化，采用油红O染色法在倒置荧光显微镜下观察细胞形态变化、脂滴的形成过程和脂滴数量变化；采用油红O提取法定量分析脂肪细胞脂滴含量变化；采用实时荧光定量PCR技术分析分化基因PPARγ和脂联素的表达变化。研究结果：在诱导分化第二天后，，染毒组剂量组率先出现脂滴。诱导分化结束后染毒剂量组脂滴含量大于空白对照组，同时不同染毒剂量组脂滴含量随染毒剂量递增。脂肪细胞分化基因PPARγ、脂联素均随着染毒剂量的增加而表达上调(P0.01)。研究结论：PAEs是脂肪细胞分化基因PPARγ的有效配体可以与之结合从而激活它，PPARγ参与脂肪细胞诱导分化过程。脂肪细胞分化基因PPARγ、脂联素均表达上调，从而使3T3-L1前体脂肪细胞变成熟脂肪细胞数量增多，脂肪颗粒明显增加。本研究结果为PAEs与肥胖的关系研究提供了一条新的思路，为相关公共卫生政策的制定提供科学依据。并为未来肥胖疾病的防治及肥胖与环境污染关系的研究开辟新途径。
[Abstract]:Objective: abnormal proliferation and differentiation of adipocytes can increase the number of adipocytes, increase the volume of adipocytes, accumulate too many fat particles, and lead to imbalance of energy metabolism, obesity and related diseases. Environmental factors are one of the influencing factors of obesity disease: phthalate ester (PAEs) is a kind of persistent organic pollutant, which has environmental estrogen-like effect. Studies on the toxicity of PAEs have been focused on reproductive toxicity. Literature reports and previous studies suggest that PAEs is associated with abdominal obesity and childhood obesity, but there is no study on the mechanism of its effects. Methods: in this study, PAEs is the peroxisome proliferator, which can activate the peroxisome proliferator receptor PPAR. The two kinds of PAESS DBP (DEHP), which were the most exposed to human body, were used to establish the experimental model of 3T3-L1 precursor adipocytes derived from mice in vitro. The 3T3-L1 precursor adipocytes were induced to differentiate into mature adipocytes by the classical hormone cocktail method. Two kinds of iconic PAEs were designed to induce the differentiation of 3T3-L1 precursor adipocytes. Oil red O staining was used to observe the morphological changes of the cells, the formation process of lipid droplets and the changes of the number of lipid droplets under inverted fluorescence microscope. The changes of lipid droplets in adipocytes were analyzed by oil red O extraction, and the expression of PPAR 纬 and adiponectin in adipocytes were analyzed by real-time fluorescence quantitative PCR. Results: after the second day of induced differentiation, lipid droplets appeared first in the dose group, and the lipid drop content in the dose group was higher than that in the blank control group after the induction of differentiation. The adipocyte differentiation gene PPAR 纬 and adiponectin were all up-regulated with the increase of dose of adipocyte differentiation gene PPAR 纬 and adiponectin. Conclusion PPAR 纬 is an effective ligand of adipocyte differentiation gene PPAR 纬, which can be activated to participate in adipocyte differentiation. The expression of adipocyte differentiation gene PPAR 纬 and adiponectin is up-regulated. As a result, 3T3-L1 precursor adipocytes increased the number of mature adipocytes and increased the number of fat particles. The results of this study provide a new idea for the study of the relationship between PAEs and obesity. It provides a scientific basis for the formulation of relevant public health policies and opens up a new way for the prevention and treatment of obesity diseases and the study of the relationship between obesity and environmental pollution in the future.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R151

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