瘦素逆转CLA诱导的小鼠代谢障碍的免疫组织化学观察

发布时间：2018-05-04 23:45

本文选题：CLA + Leptin　；参考：《河南农业大学》2015年硕士论文

【摘要】：瘦素(Leptin)是由脂肪细胞分泌的一种蛋白质激素,可以减少能量的摄入,增加能量的消耗。Leptin通过与其受体结合,通过信号转导发挥生理学作用。本实验以CLA诱导的代谢障碍的小鼠为模型,采取免疫组织化学的手段研究Leptin干预动物肝脏代谢相关基因表达的变化。在饲料中添加CLA后,成功诱导出高血糖、高血脂、脂肪肝等代谢障碍指标。实验分为对照组亚油酸组(LA组)和实验组亚油酸组实验对照组CLA+生理盐水组(CLA+S)、实验组CLA+瘦素组(CLA+L)。昆明小鼠在21d断奶后,开始饲喂添加LA和CLA的鼠粮,饲喂42d后,LA组不做处理,CLA+S组背部皮下加生理盐水缓释泵,CLA+L组背部皮下加Leptin(1mg/mL)缓释泵,作用14d后,取肝脏后做免疫组化试验。结果显示,CLA组小鼠与LA组相比,与糖代谢相关的Akt、p-AKT、FOXO1的表达量下调,而在CLA+L组,Akt、p-AKT、FOXO1的表达量上调。由此可知,CLA对这几个基因表达有所抑制,使胰岛素信号通路不能顺利的传导,从而出现高血糖和胰岛素抵抗的现象。CLA+L组与CLA+S相比,这些基因的表达升高,说明瘦素可以调节这些基因的表达从而改善体内的糖代谢紊乱。瘦素可以促进胰岛素下游信号的传递与表达而发挥调节糖代谢的作用。瘦素通过与其受体结合,通路下游的p-STAT3,CLA+L组与CLA+S相比表达量显著升高(p0.05),瘦素信号通路信号转导加强。在糖异生方面起重要作用的关键酶G-6-Pase,CLA+S组与LA组相比,表达量显著下降(p0.05),糖异生活动减弱;CLA+L组与CLA+S组相比,表达量显著升高(p0.05),糖异生活动增强。脂代谢方面,CLA+S组与LA组相比,CLA+S组SRB1的表达显著降低(p0.05),CD36的表达显著升高(p0.05)。表明CLA可以降低SRB1的表达同时可以上调CD36的表达。推测SRB1参与的胆固的逆转运活动增强,CD36表达量的升高,使长链脂肪酸的摄入增加,在肝脏内堆积从而形成脂肪肝。CLA+L组与CLA+S组相比,SRB1的表达显著升高(p0.05),下调CD36的表达。说明,SRB1参与的胆固的逆转运活动减弱,CD36介导的长链脂肪酸的摄入减少。
[Abstract]:Leptin (leptin) is a protein hormone secreted by adipocytes, which can reduce energy intake and increase energy consumption. Leptin plays a physiological role by binding to its receptor and signal transduction. In this study, CLA induced metabolic disorders in mice were used as a model to study the changes of gene expression related to liver metabolism induced by Leptin by immunohistochemical method. Hyperglycemia, hyperlipidemia, fatty liver and other metabolic disorders were successfully induced by adding CLA to feed. The experiment was divided into two groups: control group, linoleic acid group (LA group), experimental group linoleic acid group (experimental control group, CLA normal saline group) and experimental group CLA leptin group. After 21 days weaning, Kunming mice were fed with the diet supplemented with LA and CLA. After 42 days, the rats in the LA group were not treated with hypodermic saline sustained release pump on the back of the CLA group. The mice in the CLAL group were subcutaneously supplemented with Leptin 1 mg / mL. After 14 days of treatment, the mice in the control group were treated with Leptin 1 mg / mL. The liver was taken for immunohistochemistry. The results showed that compared with LA group, the expression of Akttip-AKTnFOXO1 was down-regulated in the CLA L group, but up-regulated in the CLA L group. Therefore, the expression of these genes was inhibited and the insulin signaling pathway could not be transduced smoothly. The expression of these genes in CLA-L group was higher than that in CLA S group, which resulted in hyperglycemia and insulin resistance. Leptin can regulate the expression of these genes and improve the disorder of glucose metabolism in the body. Leptin can promote the transduction and expression of insulin downstream signal and regulate glucose metabolism. By binding to its receptor, the expression of p-STAT3CLAL in the downstream of the pathway was significantly higher than that of CLA S, and the signal transduction of leptin signaling pathway was enhanced. Compared with LA group, the expression of G-6-Pasea-CLA group, the key enzyme that plays an important role in glucose heterogenesis, decreased significantly (p 0.05). The expression level of glucose heterozygosity in CLA L group was significantly higher than that in CLA S group, and that in glucose heterozygosity group was higher than that in CLA S group. In terms of lipid metabolism, the expression of SRB1 in CLAs group was significantly lower than that in LA group. The results showed that CLA could decrease the expression of SRB1 and up-regulate the expression of CD36. It was speculated that the reverse transport of gallbladder solid involved in SRB1 increased the expression of CD36, increased the intake of long chain fatty acids, accumulated in liver and formed fatty liver. Compared with CLA S group, the expression of SRB1 B1 increased significantly, and the expression of CD36 was down-regulated. It indicated that SRB1 involved in reverse transport of bile and solid decreased the intake of long chain fatty acids mediated by CD36.
【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.2

【相似文献】