PD大鼠棕色脂肪组织活性改变对脂质分解和脂肪重量减少的影响

发布时间：2018-03-08 19:35

本文选题：帕金森病　切入点：交感神经　出处：《首都医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:以中枢黑质中多巴胺能神经元缺失为主要病理特征的帕金森病(Parkinson's disease,PD)的患病率和发病率逐年上升,已成为严重影响人类健康的重要疾病之一。其临床主要表现为运动系统症状和非运动系统症状,而体重减轻是其非运动系统症状之一。探索影响PD病人体重减轻、脂肪减少的因素,是提高PD病人生活质量的关键。棕色脂肪组织(BAT)作为一个特殊的脂肪组织,与骨骼肌同源,通过介导线粒体氧化呼吸链的解偶联作用,消耗甘油三酯和自由脂肪酸参与脂质代谢,是机体非震颤性产热的主要来源。前期研究中,我们发现PD模型大鼠棕色脂肪组织的交感神经活性增强,提示BAT可能与PD病人的脂肪减少、体重减轻具有相关性。设计和方法:分别将6-OHDA和生理盐水注射到大鼠左侧前脑内侧束制作PD模型和假手术(sham)大鼠,并同时切断其中的25只PD模型大鼠和10只sham大鼠的棕色脂肪组织周围的交感神经,从而抑制棕色脂肪组织的活性。9周后分别将另外25只PD模型大鼠和10只sham大鼠置于4℃冰箱中,每天检测并记录体温。10周后,将四组大鼠麻醉、处死、并分别取出肾周、皮下、附睾和棕色脂肪组织并称重。检测棕色脂肪组织的交感神经活性标记物-酪氨酸羟化酶、功能活性标记物UCP1、β3R的表达。检测肾周、附睾、皮下脂肪组织中甘油三酯、自由脂肪酸和脂肪分解相关分子表达情况。结果:1.支配大鼠BAT的交感神经活性及BAT功能相关分子的检测酪氨酸羟化酶是经典的交感神经末梢活性标记物,通过Western blot实验发现,PD模型大鼠BAT的(Tyrosine hydroxylase)TH表达含量明显高于sham组,功能标记物(uncoupling protein 1)UCP1、分解因子(β3-adrenergic receptor)β3R的含量也明显增加。将模型置于4℃条件下9天,体温检测结果显示,PD组大鼠的体温恢复速度明显较sham组快。2.去神经大鼠BAT功能活性和体重的检测去神经组中BAT的TH和UCP1的表达水平明显较非去神经组降低,且具有统计学差异。RNA水平检测显示,去神经组BAT中β3R的表达也明显下降。由此说明,我们的去神经手术是成功的,显著降低了BAT的活性和功能。且去神经组比非去神经组的体重明显增高,在肾周脂肪组织和附睾脂肪组织中也呈现相同的趋势。3.各脂肪组织脂质分解相关因子的检测我们利用Elisa检测各WATs和BAT中的甘油三酯(triglyceride,TG)和自由脂肪酸(nonesterified fatty acid,NEFA)的含量,结果显示,与非去神经组相比,在去神经组的肾周脂肪组织(retroperitoneal WAT,r WAT)和附睾脂肪组织(epididymal WAT,e WAT)中,TG的含量升高,NEFA的含量降低。在s WAT和BAT中的变化不明显。Elisa分别检测r WAT、e WAT、s WAT的脂解相关酶PKA/P-HSL/MAGL/ATGL的水平,结果显示,在r WAT和e WAT中上述5种脂解酶的含量在去神经组中明显降低,与β3R的表达呈一致性。而s WAT中变化不明显。结论:黑质纹状体通路多巴胺能神经元的损毁,降低了对支配棕色脂肪组织交感神经高级中枢核团的抑制,间接地促进了支配棕色脂肪组织交感神经活性的增加,使BAT产热功能增强,对NEFA的利用率增加,从而促进了白色脂肪组织的分解,导致PD大鼠脂肪组织的减少和体重的减轻。
[Abstract]:Objective: to dopamine in the substantia nigra neurons lack the main pathological features of Parkinson's disease (Parkinson's, disease, PD) the prevalence and incidence increased year by year, has become one of the important diseases seriously affect human health. Its main clinical manifestations and symptoms of motion system of the non motor symptoms, and weight loss is the the non motor symptoms of PD patients. To explore the influence factors of weight loss, reduced fat, is the key to improve the quality of life of PD patients. Brown adipose tissue (BAT) as a special adipose tissue, and skeletal muscle of the same source, guide the uncoupling effect of mitochondrial respiratory chain mediated by the consumption of triglycerides and free fatty acids involved in lipid metabolism, the body is the main source of non tremor thermogenesis. In previous study, we found that enhanced sympathetic activity in brown adipose tissue of PD rats, suggesting that BAT may be with PD patients of fat reduction, weight loss is associated. Design and methods: 6-OHDA and saline was injected into the rat left medial forebrain bundle PD model (sham) and sham operation rats, and turn off the sympathetic nerve around one of the 25 PD rats and 10 sham rats, brown fat tissue, thus inhibiting the activity of brown adipose tissue after.9 weeks respectively in 25 PD rats and 10 sham rats at 4 C in the refrigerator, to detect and record the temperature every day after.10 weeks, four groups of rats were sacrificed, and anesthesia, respectively removed perirenal, epididymal and subcutaneous fat, brown tissue and weighed. Tyrosine hydroxylase activity of sympathetic nerve marker detection of brown adipose tissue, functional activity markers UCP1, 3R expression. The detection of perirenal, epididymal and subcutaneous adipose tissue triglyceride, feeling the freedom of expression of fatty acid and fat decomposition related molecules Results: 1. cases. Control of tyrosine hydroxylase activity and BAT molecular detection of sympathetic nerve function in BAT rats is sympathetic nerve activity markers classic, through Western blot experiments showed that the rat model of PD BAT (Tyrosine hydroxylase) TH expression was significantly higher than that of sham group, functional marker (uncoupling protein 1) UCP1 the decomposition factor (beta 3-adrenergic, receptor) levels of beta 3R also increased significantly. The model was placed in the temperature of 4 DEG C for 9 days, the temperature test results showed that the expression level of the rats in group PD the recovery rate of group sham was significantly higher than.2. to detect BAT activity and neural function of rats weight denervation group in BAT TH and UCP1 was significantly higher than the non denervated group decreased, and has showed significant difference in the.RNA level, the expression of 3R beta to nerve group BAT also decreased significantly. Therefore, our denervation surgery was successful, significant Reduce the activity and function of BAT. And the denervation group than in the non denervated group weight increased significantly, detected in perirenal adipose tissue and epididymal adipose tissue also showed the same trend.3. adipose tissue lipolysis related factors we use Elisa to detect the WATs and BAT in Gansu oil ester (triglyceride, TG three free fatty acid (nonesterified) and fatty acid, NEFA) display the results of the content, and non denervation group than in perirenal adipose tissue denervation group (retroperitoneal WAT, R WAT) and epididymal adipose tissue (epididymal WAT, e WAT), the content of TG increased, the content of NEFA was decreased. Changes in s WAT and BAT.Elisa were not obvious in detection of R WAT, e WAT, PKA/P-HSL/MAGL/ATGL s WAT the lipolytic enzyme level, results showed that in R WAT and E WAT in the content of the 5 kinds of lipolytic enzymes in to significantly lower nerve group, and 3R expression was Consistency. S WAT did not change significantly. Conclusion: nigrostriatal dopaminergic neuron damage reduced the inhibitory innervation of brown adipose tissue sympathetic senior central nuclei, indirectly promoting the increase of sympathetic activity of brown adipose tissue, increased BAT heat production rate increased to NEFA function. Use to promote the decomposition of white adipose tissue, lead to lessening the fat tissue of PD rats and weight.

【学位授予单位】：首都医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742.5

【相似文献】