DEHP染毒小鼠2型糖尿病模型的建立与初步研究
发布时间：2018-10-30 14:09
【摘要】：糖尿病(diabetes mellitus, DM)是一种多病因的慢性代谢紊乱疾病,是由于胰岛素分泌减少、功能减退或者胰岛素抵抗相关等因素导致高血糖含量,从而引起糖、脂肪和蛋白质代谢紊乱以及逐步造成其他器官的损伤和一系列功能障碍。糖尿病是当今世界最常见的慢性病症之一,被列为继心血管疾病和肿瘤之后的第三大顽固疾病。目前中国已成为世界上糖尿病患者最多的国家,其中2型糖尿病的发病率最高。DEHP作为应用广泛的环境污染物,其毒性研究已为人们所重视。美国Stahlhut的流行病学研究发现,糖尿病与邻苯二甲酸酯类化合物之间存在着相关性,但目前还没有相应的动物模型来支持这一假说,同时邻苯二甲酸酯类化合物对糖尿病影响的机制通路也尚未明确。因此本实验通过采用DEHP染毒暴露的Balb/C小鼠经高脂高糖饲养后注射STZ来诱导制备2型糖尿病模型,以达到研究DEHP与糖尿病内在联系的目的,为进一步研究受环境因素影响的糖尿病发病机制提供基础的分子学理论。 1. DEHP联合效应诱导的糖尿病模型的建立 小鼠买回后适应性饲养3-4天后随机分组,单数组喂养高脂高糖饲料,双数组喂养普通饲料(包括0组),每天0组和1组进行吐温-80+生理盐水灌胃、2组和3组进行10mg/kg/d DEHP灌胃染毒、4组和5组进行50mg/kg/d DEHP灌胃染毒、6组和7组进行250mg/kg/d DEHP灌胃染毒。染毒持续3周后,禁食12h,尾部取血测空腹血糖,喂食,再次禁食12h,第二天按体重腹腔注射100mg/kgSTZ溶液(所有对照组注射柠檬酸缓冲液)。1周后测血糖值,3天后再次禁食12h,第二天注射80mg/kg STZ溶液,1周后测血糖,出现糖尿病小鼠,不再注射STZ,恢复灌胃,再持续3周,血糖值基本稳定后,处死,测定肝脏氧化损伤指标、血清胰岛素含量以及胰腺和肾脏组织切片等。 2. DEHP对糖尿病发生的直接和间接影响 本实验通过DEHP单独暴露组和DEHP染毒、高脂饲养、链尿佐菌素注射联合诱导的糖尿病小鼠模型组的建立,对DEHP在DEHP联合作用诱导的糖尿病发病过程中所产生的直接和间接作用进行了对照研究。实验结果显示DEHP作为一种环境因子,确实能对糖尿病的发生产生影响。结合所测的实验指标,可以进行推断,单独的DEHP灌胃染毒可以对小鼠机体产生相应的氧化损伤以及各种脏器毒性效应,这些刺激使得单独灌胃组的数据与空白对照组对比呈现出一定的差异,并且这种差异与DEHP浓度存在着剂量相关性。但是DEHP的毒性作用不能直接诱导糖尿病的发生,而在联合作用诱导的建模组,DEHP染毒暴露发挥了明显的效应,这从建模组小鼠的糖尿病症状的出现情况以及胰岛素含量的测定结果可以得到证实。此外氧化损伤指标的结果也反映出DEHP的辅助作用,同时还可以进一步推测DEHP对糖尿病的影响机制并不是完全直接通过其氧化损伤作用介导,应该还有其他诸如生殖毒性(睾酮影响)、促炎症作用等效应通路使得糖尿病的发生易化。但这一推论还需要通过对联合效应建模组和单独DEHP灌胃染毒组小鼠体内各种与胰岛素抗性以及胰岛素作用通路相关的细胞因子以及糖尿病易感基因的测定和研究后进行进一步的推导。
[Abstract]:Diabetes mellitus (DM) is a multi-etiopathogenic chronic metabolic disorder, which is caused by decreased insulin secretion, hypofunction or insulin resistance, resulting in sugar, Disorders of fat and protein metabolism and progressive damage to other organs and a series of dysfunction. Diabetes is one of the most common chronic conditions in the world and is listed as the third major stubborn disease following cardiovascular disease and cancer. China has become the most diabetic in the world, with the highest incidence of type 2 diabetes. As an environmental pollutant, DEHP has been paid more attention to the toxicity of DEHP. The epidemiological study of Stahlmut in USA has found that there is a correlation between diabetes and phthalates, but there is no corresponding animal model to support this hypothesis, and the mechanism pathway of phthalate esters on diabetes is not yet clear. Therefore, the model of type 2 diabetes mellitus was induced by injecting STZ after high fat and high sugar feeding in the mice exposed by DEHP, so as to reach the purpose of studying the relationship between DEHP and diabetes. To further study the molecular theory which provides the basis for the pathogenesis of diabetes affected by environmental factors. 1. DEHP combined effect-induced diabetic model After 3-4 days after the establishment of the model, the mice were fed with high-fat and high-sugar feed, and the double-array fed common feed. The dosage of Tween-80 + normal saline was carried out in group 0 and group 1 in group 0 and group 1, group 2 and group 3 were treated with 10mg/ kg/ d DEHP by gavage, group 4 and group 5 were treated with 50mg/ kg/ d DEHP, and treated with 250mg/ kg/ d D in 6 groups and 7 groups respectively. EHP was administered by gavage. After 3 weeks of exposure, fasting for 12h, fasting blood glucose was taken at the tail, fed and fasted again for 12h, and 100 mg/ kg STZ solution was injected intraperitoneally on the next day (citric acid buffer was injected in all control groups). After 1 week, blood glucose level was measured, and after 3 days, After fasting for 12h, the 80mg/ kg STZ solution was injected the next day, blood glucose was measured after 1 week, diabetic mice were observed, STZ was no longer injected, gastric administration was resumed for 3 weeks, blood glucose level was basically stable, sacrificed, liver oxidative damage index, serum insulin content and pancreas were determined. Renal tissue slices et al. 2. DEHP vs. sugar The direct and indirect effects of urinary disease were studied by DEHP alone exposure group and DEHP infection, high fat feeding, streptozoin injection linkage. The establishment of the induced diabetic mouse model group and the effect of DEHP on the pathogenesis of diabetes induced by DEHP combined action The direct and indirect effects of DEHP were studied. The experimental results show that DEHP is an environmental factor. According to the experimental indexes, it can be concluded that the individual DEHP can produce the corresponding oxidative damage and various organ toxicity effects on the body of the mice, and the stimulation makes the data of the single oral gavage group and The contrast of the blank control group showed a certain difference, and the difference There was a dose-related relationship with DEHP concentration. However, DEHP's toxic effect could not directly induce the occurrence of diabetes, while in the modeling group induced by joint action, DEHP was exposed to an obvious effect, which was from the occurrence of diabetic symptoms in the modeling group and the pancreas. In addition, the results of oxidative damage index also reflect DEHP's auxiliary function, and it is also possible to speculate that DEHP's mechanism of influence on diabetes is not mediated directly by its oxidative damage, and it should also There are other effects such as reproductive toxicity (testosterone), pro-inflammatory action, The effects of these pathways make the onset of diabetes easy. However, this inference also requires the use of cytokines as well as diabetic susceptibility genes associated with insulin resistance and insulin action pathways in mice treated with a combination effect modeling group and a separate DEHP gavage group.
【学位授予单位】：华中师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R587.1;R-332
本文编号：2300221
[Abstract]:Diabetes mellitus (DM) is a multi-etiopathogenic chronic metabolic disorder, which is caused by decreased insulin secretion, hypofunction or insulin resistance, resulting in sugar, Disorders of fat and protein metabolism and progressive damage to other organs and a series of dysfunction. Diabetes is one of the most common chronic conditions in the world and is listed as the third major stubborn disease following cardiovascular disease and cancer. China has become the most diabetic in the world, with the highest incidence of type 2 diabetes. As an environmental pollutant, DEHP has been paid more attention to the toxicity of DEHP. The epidemiological study of Stahlmut in USA has found that there is a correlation between diabetes and phthalates, but there is no corresponding animal model to support this hypothesis, and the mechanism pathway of phthalate esters on diabetes is not yet clear. Therefore, the model of type 2 diabetes mellitus was induced by injecting STZ after high fat and high sugar feeding in the mice exposed by DEHP, so as to reach the purpose of studying the relationship between DEHP and diabetes. To further study the molecular theory which provides the basis for the pathogenesis of diabetes affected by environmental factors. 1. DEHP combined effect-induced diabetic model After 3-4 days after the establishment of the model, the mice were fed with high-fat and high-sugar feed, and the double-array fed common feed. The dosage of Tween-80 + normal saline was carried out in group 0 and group 1 in group 0 and group 1, group 2 and group 3 were treated with 10mg/ kg/ d DEHP by gavage, group 4 and group 5 were treated with 50mg/ kg/ d DEHP, and treated with 250mg/ kg/ d D in 6 groups and 7 groups respectively. EHP was administered by gavage. After 3 weeks of exposure, fasting for 12h, fasting blood glucose was taken at the tail, fed and fasted again for 12h, and 100 mg/ kg STZ solution was injected intraperitoneally on the next day (citric acid buffer was injected in all control groups). After 1 week, blood glucose level was measured, and after 3 days, After fasting for 12h, the 80mg/ kg STZ solution was injected the next day, blood glucose was measured after 1 week, diabetic mice were observed, STZ was no longer injected, gastric administration was resumed for 3 weeks, blood glucose level was basically stable, sacrificed, liver oxidative damage index, serum insulin content and pancreas were determined. Renal tissue slices et al. 2. DEHP vs. sugar The direct and indirect effects of urinary disease were studied by DEHP alone exposure group and DEHP infection, high fat feeding, streptozoin injection linkage. The establishment of the induced diabetic mouse model group and the effect of DEHP on the pathogenesis of diabetes induced by DEHP combined action The direct and indirect effects of DEHP were studied. The experimental results show that DEHP is an environmental factor. According to the experimental indexes, it can be concluded that the individual DEHP can produce the corresponding oxidative damage and various organ toxicity effects on the body of the mice, and the stimulation makes the data of the single oral gavage group and The contrast of the blank control group showed a certain difference, and the difference There was a dose-related relationship with DEHP concentration. However, DEHP's toxic effect could not directly induce the occurrence of diabetes, while in the modeling group induced by joint action, DEHP was exposed to an obvious effect, which was from the occurrence of diabetic symptoms in the modeling group and the pancreas. In addition, the results of oxidative damage index also reflect DEHP's auxiliary function, and it is also possible to speculate that DEHP's mechanism of influence on diabetes is not mediated directly by its oxidative damage, and it should also There are other effects such as reproductive toxicity (testosterone), pro-inflammatory action, The effects of these pathways make the onset of diabetes easy. However, this inference also requires the use of cytokines as well as diabetic susceptibility genes associated with insulin resistance and insulin action pathways in mice treated with a combination effect modeling group and a separate DEHP gavage group.
【学位授予单位】：华中师范大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R587.1;R-332
【参考文献】
相关期刊论文 前2条
1 宋慧婷;吴振斌;贺锋;成水平;梁威;张丽萍;;武汉月湖和莲花湖表层沉积物中持久性有机物的污染状况[J];长江流域资源与环境;2008年03期
2 王家文;杜琪珍;宋英琦;;塑料工业区附近农田蔬菜DEHP的浓度水平及评价[J];环境科学;2010年10期
，本文编号：2300221
本文链接：https://www.wllwen.com/xiyixuelunwen/2300221.html
最近更新
教材专著
