丁苯酞对LPS腹腔注射诱导帕金森病小鼠模型的保护作用

发布时间：2018-05-12 16:00

本文选题：帕金森 + 丁苯酞　；参考：《蚌埠医学院》2017年硕士论文

【摘要】：目的:1.采用腹腔注射LPS法诱导C57BL/6小鼠帕金森病模型,从病因学角度模拟帕金森病自然发病过程;2.探讨丁苯酞对该模型小鼠的神经保护作用及可能机制。方法:将60只C57BL/6小鼠随机分为空白组(Control)、模型组(LPS)、治疗组(LPS+NBP)。模型组给予单次腹腔注射LPS(5mg/kg)诱导帕金森病模型;治疗组在建模的同时(具体步骤同模型组)给予丁苯酞(120mg/kg,溶于1ml植物油)灌胃治疗,每天一次,持续30天;空白对照组在对应的时间给予相等体积生理盐水腹腔注射。1.分别于实验开始后的第30天、第90天、第150天和第210天,在空白组与模型组各随机取出6只小鼠,采用转棒实验及旷场试验评估小鼠行为变化情况,完成行为学观测后麻醉小鼠,心脏固定液灌流,取脑组织,免疫组织化学染色法观察小鼠炓质区酪氨酸羟化酶(TH)阳性细胞、IBA-1阳性细胞(活化的小胶质细胞)数目的变化,观察炎性因子IL-1β的表达情况。2.分别于实验开始后的第210天,取空白组、模型组、治疗组每组各6只小鼠行转棒实验及旷场实验,完成行为学观察后麻醉小鼠,每组取三只小鼠心脏固定液灌流,取脑组织,免疫组织化学染色法观察小鼠炓质TH、IBA-1、IL-1β、CHOP的表达情况,另三只小鼠取中脑炓质部位,用蛋白质印迹法(Western blot)测定P-JNK、P-P38及P-ERK的表达水平的。结果:1.单次LPS腹腔注射能够能诱发C57BL/6小鼠行为学障碍:与对照组相比,模型组小鼠的运动能力在第30天并未表现出明显变化(P0.01),但从第90天起模型组小鼠表现出明显的运动能力下降(P0.01),差异有统计学意义,并逐渐表现出震颤、竖毛等PD症状,模型组小鼠提前出现毛色变白、胡须脱落等老化现象。2.单次LPS腹腔注射能够诱发C57BL/6小鼠长期脑内炎症反应:与对照组相比,模型组小鼠中脑黑质区IBA-1、IL-1β阳性细胞数明显增多(P0.01),差异有统计学意义。3.单次LPS腹腔注射能够诱发C57BL/6小鼠多巴胺能神经元丢失:与对照组相比,模型组小鼠中脑黑质区TH阳性神经元细胞树突减少,细胞染色变浅,细胞数目明显下降(P0.01),差异有统计学意义。4.丁苯酞可以缓解帕金森病模型小鼠的行为学障碍:与模型组相比,治疗组小鼠表现出较强的运动能力(P0.01),5.丁苯酞可以缓解帕金森病模型小鼠的多巴胺能神经元损伤:与模型组相比,治疗组小鼠炓质TH阳性细胞形态趋于正常,神经元凸起增多,细胞数量明显增加(P0.01)。6.丁苯酞可以缓解帕金森病模型小鼠脑内炎症反应:与模型组相比,治疗组小鼠炓质区IBA-1、IL-1β阳性细胞数量明显降低(P0.01),7.丁苯酞可以影响帕金森病模型小鼠MAPK通路的激活:与对照组相比,模型组小鼠炓质区P-JNK、P-P38的表达水平明显升高(P0.01);与模型组相比治疗组的P-JNK、P-P38的表达水平出现显著的下降(P0.01);而三组小鼠P-ERK的水平未见明显异常(P0.01)。8.丁苯酞可以改善帕金森病模型小鼠炓质区内质网应激:与对照组相比,模型组小鼠炓质区CHOP的表达水平明显升高(P0.01),治疗组小鼠黑质区CHOP的表达水平未见明显差异(P0.01)结论:1.单次给予腹腔注射LPS能够成功诱导慢性帕金森病小鼠模型,其发病机制可能与长期持续性的脑内炎症反应有关。2.丁苯酞对LPS腹腔注射诱导帕金森病模型小鼠具有神经保护作用,该保护机制可能与缓解脑内慢性炎症反应、降低MAPK通路的激活及改善内质网应激有关。
[Abstract]:Objective: 1. the model of Parkinson's disease in C57BL/6 mice was induced by intraperitoneal injection of LPS, and the natural pathogenesis of Parkinson's disease was simulated from the etiological point of view; and 2. to explore the neuroprotective effect and possible mechanism of butylphthalide on the model mice. Methods: 60 C57BL/6 mice were randomly divided into blank group (Control), model group (LPS), and treatment group (LPS+NBP). The group was given a single intraperitoneal injection of LPS (5mg/kg) to induce Parkinson's disease model, and the treatment group was given the treatment of butylphthalide (120mg/kg, soluble in 1ml plant oil) at the same time (120mg/kg, 1ml plant oil) at the same time (the model group). The control group was given the same volume of normal saline at the corresponding time and intraperitoneal injection of.1. at the beginning of the experiment. After thirtieth days, ninetieth days, 150th days and 210th days, 6 mice were randomly removed from the blank group and the model group. The change of mice behavior was evaluated by the rod experiment and the open field test. After the behavior observation, the anesthetized mice were perfused, the brain tissue was taken and the immunohistochemical staining method was used to observe the tyrosine hydroxylase in the mice. TH) positive cells, the number of IBA-1 positive cells (activated microglia), and observe the expression of IL-1 beta of inflammatory factors,.2., 210th days after the start of the experiment, and take the blank group, model group, 6 mice in each group of each group in each group of the treatment group. After the behavior observation, the anesthetized mice were completed, and three mice in each group were taken. The expression of TH, IBA-1, IL-1 beta and CHOP was observed in the brain tissue by immunohistochemistry. The expression level of P-JNK, P-P38 and P-ERK in the other three mice was measured by the Western blot method (Western blot). Results: 1. single LPS intraperitoneal injection could induce the behavioral barrier of C57BL/6 mice. Hinder: compared with the control group, the exercise ability of the model mice did not show significant changes at thirtieth days (P0.01), but from the ninetieth day, the mice in the model group showed significant movement ability (P0.01), and the difference was statistically significant, and the PD symptoms such as tremor, erect hair and other symptoms were gradually displayed. Aging phenomenon.2. single LPS intraperitoneal injection can induce long-term intracerebral inflammatory response in C57BL/6 mice: compared with the control group, the number of IL-1 beta positive cells in the middle brain of the model group was significantly increased (P0.01), and the difference was statistically significant in.3. single LPS intraperitoneal injection to induce the loss of dopaminergic neurons in the C57BL/6 mice: with the control group. In the model group, the dendrites of TH positive neurons in the mesencephalic area of the mice decreased, the cell dyed light and the number of cells decreased significantly (P0.01). The difference was statistically significant that the.4. butylphthalide could relieve the behavioral disorders of the Parkinson's disease model mice. Compared with the model group, the mice showed strong exercise ability (P0.01), and 5. butylphthalide could be found in the model group. To alleviate the dopaminergic neuron damage in Parkinson's disease model mice: compared with the model group, the morphology of the TH positive cells in the treatment group tended to be normal, the neuron bulge increased and the number of cells increased significantly (P0.01).6. butylphthalide could alleviate the inflammation in the brain of the Parkinson's disease model mice. In area IBA-1, the number of IL-1 beta positive cells decreased significantly (P0.01). 7. butylphthalide could affect the activation of MAPK pathway in Parkinson's disease model mice. Compared with the control group, the expression level of P-JNK and P-P38 in the model group was significantly increased (P0.01), and the expression level of P-P38 in the treatment group was significantly lower than that in the model group (P0.01). There was no obvious abnormality in the level of P-ERK in the three groups (P0.01).8. butylphthalide could improve the endoplasmic reticulum stress in the Parkinson's disease model mice. Compared with the control group, the expression level of CHOP was significantly increased in the model group (P0.01). The level of CHOP in the substantia nigra area of the treatment group was not significantly different (P0.01): the 1. single abdomen was given to the abdomen. Intraperitoneal injection of LPS can successfully induce a mouse model of chronic Parkinson's disease. The pathogenesis may be related to the long-term persistent intracerebral inflammation reaction related to the neuroprotective effect of.2. butylphthalide on LPS induced Parkinson's disease model mice induced by intraperitoneal injection. The protective mechanism may be associated with the response to slow inflammation in the brain and the reduction of the activation and modification of the MAPK pathway. Good endoplasmic reticulum stress is related.

【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742.5;R-332

【相似文献】