铅和高脂饮食暴露对中枢神经系统炎性微环境的影响及其机制研究

发布时间：2018-05-06 15:12

本文选题：铅 + 高脂饮食　；参考：《华北理工大学》2017年硕士论文

【摘要】：目的本研究从脉络丛调控免疫细胞进入中枢神经系统的角度探讨在铅和高脂饮食条件下中枢神经系统炎性微环境的变化的机制,以期为铅和高脂饮食暴露致神经损伤的生物标志物的研究提供新的靶点。方法1实验动物处理及分组:60只健康C57BL/6J雄性小鼠,随机分为对照组,高脂饮食组,铅暴露组,铅暴露+高脂饮食组。采用自由饮水的方式染毒,染毒时间为12周。对照组予基础饲料喂养,高脂饮食组予高脂饲料喂养,铅暴露组予基础饲料喂养同时饮用质量浓度为300mg/L的醋酸铅饮用水;铅暴露+高脂饮食组予饮用质量浓度为300 mg/L的醋酸铅饮用水同时予高脂饲料喂养。2应用Morris水迷宫、新物体识别实验及旷场试验对小鼠进行神经行为测试。3应用ICP-MS法检测小鼠海马组织中铅元素的含量。4应用ELISA法和试剂盒检测小鼠血清中总胆固醇(T-CHO)、甘油三酯(TG)的含量和海马中炎症因子及BDNF含量的变化。5应用流式细胞术检测血液、脾脏中Th1/CD4+、Th2/CD4+、Th17/CD4+百分率的变化。6应用real-time PCR检测小鼠脉络丛中粘连蛋白VCAM-1、ICAM-1的m RNA表达的变化。计量资料的统计学描述以-x±s表示,不同组别间比较应用方差分析和两两比较的LSD检验,P0.05具有统计学意义。结果1.实验小鼠生长发育情况:各组小鼠体重随时间延长均升高,给予高脂饮食小鼠体重显著高于普通饲料喂养小鼠(P0.05)。2.铅和高脂饮食暴露对小鼠血清中总胆固醇(T-CHO)、甘油三酯(TG)的影响:高脂饮食小鼠血液中总胆固醇(T-CHO)、甘油三酯(TG)含量升高(P0.05);未见铅暴露+高脂饮食组小鼠血清中T-CHO、TG含量较高脂饮食组有明显变化。3.铅和高脂饮食暴露对小鼠神经行为功能的影响:高脂饮食组、铅暴露组、铅暴露+高脂饮食组小鼠的潜伏期高于对照组,穿台次数下降(P0.05)。与高脂饮食组及铅暴露组相比,铅暴露+高脂饮食组第3天潜伏期明显延长,穿台次数减少(P0.05)。与对照组比较,高脂饮食组、铅暴露组、铅暴露+高脂饮食组小鼠中央区域路程及站立次数均减少(P0.05);铅暴露+高脂饮食组小鼠中央区域路程较高脂饮食组或铅暴露组明显下降,差异有统计学意义(P0.05)4.铅和高脂饮食暴露对小鼠海马组织中铅含量的影响:铅暴露组、铅暴露+高脂饮食组小鼠海马中的铅含量分别为对照组的8.8倍、9.6倍;与高脂饮食组相比,铅暴露组、铅暴露+高脂饮食组小鼠海马中的铅含量均增加(P0.05)。5.铅和高脂饮食暴露对海马中炎症因子及BDNF的影响:高脂饮食或铅暴露小鼠海马中炎症因子IL-17、IL-6、IL-1b、IFN-g水平高于对照组,铅暴露和高脂饮食联合暴露小鼠海马中IL-17、IL-1b的水平高于单独暴露组(P0.05)。与对照组相比,高脂饮食组、铅暴露组BDNF含量均下降;未见铅暴露+高脂饮食组海马中BDNF含量较高脂饮食组或铅暴露组有明显差异。6.铅和高脂饮食暴露对小鼠血清中炎症因子的影响:与对照组比较,高脂饮食组、铅暴露组、铅暴露+高脂饮食组血清中IL-17含量增加,TGF-β1含量下降(P0.05)。铅暴露+高脂饮食组血清中IL-17含量较高脂饮食组或铅暴露组明显升高(P0.05)。7.铅和高脂饮食暴露对小鼠血液、脾脏中Th1/CD4+、Th2/CD4+、Th17/CD4+百分率的影响:高脂饮食组、铅暴露组、铅暴露+高脂饮食组血液中Th1/CD4+的百分率分别为2.10%、3.48%、4.18%,高于对照组的1.74%。Th17/CD4+的百分率分别为1.42%、1.46%、1.56%;高于对照组的0.67%。高脂饮食组、铅暴露组、铅暴露+高脂饮食组脾脏中Th1/CD4+的百分率分别为11.16%、13.23%、12.11%,较对照组的6.90%明显升高。Th17/CD4+的百分率分别为3.58%、3.78%、4.18%,高于对照组的2.62%;铅暴露+高脂饮食组血液、脾脏中Th2/CD4+的百分率显著降低(P0.05)。分别与高脂饮食组和铅暴露相比较,铅暴露+高脂饮食组血液中Th1/CD4+的百分率升高,血液及脾脏中Th2/CD4+的百分率降低(P0.05)。8.铅和高脂饮食暴露对小鼠脉络丛中粘连蛋白VCAM-1、ICAM-1 m RNA表达影响:对照组比较,铅暴露、高脂饮食可导致小鼠脉络丛中VCAM-1、ICAM-1的m RNA表达增加(P0.05);与高脂饮食组和铅暴露组相比,铅暴露+高脂饮食组VCAM-1的m RNA表达均显著上调(P0.05)。结论铅暴露和高脂饮食均可导致小鼠认知功能所伤,学习记忆能力和探究能力下降,且铅与高脂饮食联合暴露可加剧神经损伤,这可能与外周血中的免疫细胞及脉络丛的免疫监视功能失调,引起中枢神经系统炎性微环境的改变有关。
[Abstract]:Objective to explore the mechanism of the changes in the inflammatory microenvironment of the central nervous system under the lead and high fat diet, in order to provide new targets for the study of biomarkers in lead and high fat diet exposure, in order to provide a new target for the study of biomarkers in lead and high fat diet exposure. Methods 1 experimental animals were treated and divided into groups: 60 Only healthy C57BL/6J male mice were randomly divided into control group, high fat diet group, lead exposure group, lead exposure + high fat diet group. The free drinking water was used for 12 weeks. The control group was fed with basic diet, high fat diet group was fed with high fat diet, and the lead exposure group was given the quality of basic diet for 300mg/L. Lead acetate drinking water; lead exposure + high fat diet group give drinking water with a drinking mass of 300 mg/L of lead acetate drinking water and high fat diet feeding.2 Morris water maze; new object identification experiment and open field test on mice neurobehavioral test.3 application ICP-MS method to detect the content of lead in hippocampus of mice.4 application ELISA method The changes of total cholesterol (T-CHO), triglyceride (TG), inflammatory factors and BDNF content in the hippocampus were detected by the kit and kits..5 applied flow cytometry to detect the blood and the changes of Th1/CD4+, Th2/CD4+, Th17/CD4+ percentage in the spleen. Real-time PCR was used to detect the VCAM-1 of the adhesion protein in the choroid series of mice. The statistical description of the measurement data was expressed as -x + s, and the variance analysis and the 22 comparison LSD test were used among the different groups. The growth and development of the 1. experimental mice were increased. The weight of the mice in each group increased with time, and the weight of the mice was significantly higher than that of the normal feed mice (P 0.05) the effect of.2. lead and high fat diet exposure on serum total cholesterol (T-CHO) and triglyceride (TG) in mice: high fat diet mice blood total cholesterol (T-CHO), triglyceride (TG) content increased (P0.05); no lead exposure + high fat diet group of mice serum T-CHO, TG content high fat diet group has obvious changes of.3. lead and high fat diet exposure The effects on the neurobehavioral function of mice: high fat diet group, lead exposure group, lead exposure + high fat diet group were higher than the control group, the number of wear stage decreased (P0.05). Compared with the high fat diet group and lead exposure group, the third day latency of lead exposure + high fat diet group was obviously prolonged and the number of table wearing decreased (P0.05). In the diet group, lead exposure group, lead exposure + high fat diet group, the central area and the number of standing were decreased (P0.05); lead exposure + high fat diet group had a significant decrease in the central area of the central region of the high fat diet group or lead exposure group. The difference was statistically significant (P0.05) the effect of lead and high fat diet exposure on the lead content in the hippocampus of mice: Lead Lead exposure in the hippocampus of exposure group, lead exposure + high fat diet group was 8.8 times and 9.6 times as high as that of the control group. Lead exposure group, lead exposure group, lead exposure + high fat diet group increased the lead content in hippocampus of mice (P0.05).5. lead and high fat diet exposure on the hippocampus inflammatory factors and BDNF: high fat diet or lead exposure small exposure The levels of inflammatory factors IL-17, IL-6, IL-1b, and IFN-g in the hippocampus of rats were higher than those in the control group. The level of IL-17 and IL-1b in the hippocampus of mice exposed to lead exposure and high fat diet was higher than that of the single exposure group (P0.05). Compared with the control group, the levels of BDNF in the lead exposed group and the high fat diet group were lower than those in the control group, and there was no higher BDNF content in the hippocampus of the lead exposed and high fat diet group. The effects of.6. lead exposure and high fat diet exposure on the serum inflammatory factors in mice were significantly different. Compared with the control group, the content of IL-17 in the serum of high fat diet group, lead exposure group, lead exposure + high fat diet group increased, TGF- beta 1 decreased (P0.05). The IL-17 content in the serum of lead exposure + high fat diet group was higher than that of fat diet group or lead group The exposure group significantly increased the percentage of Th1/CD4+, Th2/CD4+ and Th17/CD4+ in the blood and spleen of mice (P0.05).7. lead and high fat diet exposure: the percentage of Th1/CD4+ in the blood of high fat diet group, lead exposure group, lead exposure + high fat diet group was 2.10%, 3.48%, 4.18% respectively, and the percentage of 1.74%.Th17/CD4+ in the control group was 1.42%, 1.4 respectively. 6%, 1.56%; the percentage of Th1/CD4+ in the 0.67%. high fat diet group, lead exposure group, lead exposure + high fat diet group was 11.16%, 13.23%, 12.11% respectively, compared with the control group, the percentage of.Th17/CD4+ was 3.58%, 3.78%, 4.18%, higher than the control group 2.62%, lead exposure + high fat diet group blood, Th2/CD4 in spleen. The percentage of + is significantly lower (P0.05). Compared with the high fat diet group and the lead storm, the percentage of Th1/CD4+ in the blood and the spleen in the high fat diet group increased, the percentage of Th2/CD4+ in the blood and spleen decreased (P0.05).8. lead and high fat diet exposure on the VCAM-1 and ICAM-1 m RNA expression in the choroid series of mice: comparison group, Lead exposure, high fat diet can lead to VCAM-1 and ICAM-1 m RNA expression in mouse choroid clusters (P0.05). Compared with high fat diet group and lead exposure group, the m RNA expression of VCAM-1 in lead exposure + high fat diet group is significantly up (P0.05). Conclusion lead exposure and high fat diet can lead to cognitive impairment, learning and memory ability and exploration ability in mice. Decrease, and combined exposure of lead and high fat diet can aggravate nerve damage, which may be associated with the immune monitoring of the immune cells and choroid plexus in peripheral blood, which may cause changes in the inflammatory microenvironment of the central nervous system.

【学位授予单位】：华北理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R114

【参考文献】