DEHP染毒小鼠2型糖尿病模型的建立與初步研究
發(fā)布時間：2018-10-30 14:09
【摘要】：糖尿病(diabetes mellitus, DM)是一種多病因的慢性代謝紊亂疾病,是由于胰島素分泌減少、功能減退或者胰島素抵抗相關(guān)等因素導(dǎo)致高血糖含量,從而引起糖、脂肪和蛋白質(zhì)代謝紊亂以及逐步造成其他器官的損傷和一系列功能障礙。糖尿病是當(dāng)今世界最常見的慢性病癥之一,被列為繼心血管疾病和腫瘤之后的第三大頑固疾病。目前中國已成為世界上糖尿病患者最多的國家,其中2型糖尿病的發(fā)病率最高。DEHP作為應(yīng)用廣泛的環(huán)境污染物,其毒性研究已為人們所重視。美國Stahlhut的流行病學(xué)研究發(fā)現(xiàn),糖尿病與鄰苯二甲酸酯類化合物之間存在著相關(guān)性,但目前還沒有相應(yīng)的動物模型來支持這一假說,同時鄰苯二甲酸酯類化合物對糖尿病影響的機制通路也尚未明確。因此本實驗通過采用DEHP染毒暴露的Balb/C小鼠經(jīng)高脂高糖飼養(yǎng)后注射STZ來誘導(dǎo)制備2型糖尿病模型,以達到研究DEHP與糖尿病內(nèi)在聯(lián)系的目的,為進一步研究受環(huán)境因素影響的糖尿病發(fā)病機制提供基礎(chǔ)的分子學(xué)理論。 1. DEHP聯(lián)合效應(yīng)誘導(dǎo)的糖尿病模型的建立 小鼠買回后適應(yīng)性飼養(yǎng)3-4天后隨機分組,單數(shù)組喂養(yǎng)高脂高糖飼料,雙數(shù)組喂養(yǎng)普通飼料(包括0組),每天0組和1組進行吐溫-80+生理鹽水灌胃、2組和3組進行10mg/kg/d DEHP灌胃染毒、4組和5組進行50mg/kg/d DEHP灌胃染毒、6組和7組進行250mg/kg/d DEHP灌胃染毒。染毒持續(xù)3周后,禁食12h,尾部取血測空腹血糖,喂食,再次禁食12h,第二天按體重腹腔注射100mg/kgSTZ溶液(所有對照組注射檸檬酸緩沖液)。1周后測血糖值,3天后再次禁食12h,第二天注射80mg/kg STZ溶液,1周后測血糖,出現(xiàn)糖尿病小鼠,不再注射STZ,恢復(fù)灌胃,再持續(xù)3周,血糖值基本穩(wěn)定后,處死,測定肝臟氧化損傷指標、血清胰島素含量以及胰腺和腎臟組織切片等。 2. DEHP對糖尿病發(fā)生的直接和間接影響 本實驗通過DEHP單獨暴露組和DEHP染毒、高脂飼養(yǎng)、鏈尿佐菌素注射聯(lián)合誘導(dǎo)的糖尿病小鼠模型組的建立,對DEHP在DEHP聯(lián)合作用誘導(dǎo)的糖尿病發(fā)病過程中所產(chǎn)生的直接和間接作用進行了對照研究。實驗結(jié)果顯示DEHP作為一種環(huán)境因子,確實能對糖尿病的發(fā)生產(chǎn)生影響。結(jié)合所測的實驗指標,可以進行推斷,單獨的DEHP灌胃染毒可以對小鼠機體產(chǎn)生相應(yīng)的氧化損傷以及各種臟器毒性效應(yīng),這些刺激使得單獨灌胃組的數(shù)據(jù)與空白對照組對比呈現(xiàn)出一定的差異,并且這種差異與DEHP濃度存在著劑量相關(guān)性。但是DEHP的毒性作用不能直接誘導(dǎo)糖尿病的發(fā)生,而在聯(lián)合作用誘導(dǎo)的建模組,DEHP染毒暴露發(fā)揮了明顯的效應(yīng),這從建模組小鼠的糖尿病癥狀的出現(xiàn)情況以及胰島素含量的測定結(jié)果可以得到證實。此外氧化損傷指標的結(jié)果也反映出DEHP的輔助作用,同時還可以進一步推測DEHP對糖尿病的影響機制并不是完全直接通過其氧化損傷作用介導(dǎo),應(yīng)該還有其他諸如生殖毒性(睪酮影響)、促炎癥作用等效應(yīng)通路使得糖尿病的發(fā)生易化。但這一推論還需要通過對聯(lián)合效應(yīng)建模組和單獨DEHP灌胃染毒組小鼠體內(nèi)各種與胰島素抗性以及胰島素作用通路相關(guān)的細胞因子以及糖尿病易感基因的測定和研究后進行進一步的推導(dǎo)。
[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.
【學(xué)位授予單位】：華中師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R587.1;R-332
本文編號：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.
【學(xué)位授予單位】：華中師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R587.1;R-332
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