Th17和MDSC在帕金森病发生发展中的作用及机制

发布时间：2018-02-03 19:50

本文关键词： 帕金森病 1-甲基-4-苯基-1 2 3 6-四氢吡啶辅助性T细胞17 髓系抑制性细胞神经炎性　出处：《江苏大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:通过对辅助性T细胞17(Th17)和髓系抑制性细胞(MDSC)在初诊帕金森病(PD)患者、PD小鼠模型中表达及相关性的检测,探究Th17和MDSC在PD病程中的表达情况并阐明二者在PD中的作用机制。方法:1、收集临床上初诊且未行相关抗PD治疗的PD患者32例作为PD组,采用Hoehr-Yahr分级和统一帕金森病评定量表评价患者病情的严重程度,并选入32名同年龄段体检正常的健康志愿者作为对照组。流式细胞仪检测两组人员外周血Th17和MDSC的细胞百分率,并分析Th17、MDSC和PD病情严重程度间的相关性。2、选择C57BL/6小鼠40只随机平均分为PD组、PD+吉西他滨组、吉西他滨组和对照组。PD组和PD+吉西他滨组小鼠予1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)建立PD模型,PD+吉西他滨组和吉西他滨组小鼠另予吉西他滨腹腔注射,对照组小鼠予相同剂量生理盐水腹腔注射。通过小鼠一般运动情况(震颤、运动、步态、毛发等表现)、行为学实验(爬杆实验、悬挂实验、强迫游泳实验)、免疫组化检测中脑黑质区酪氨酸羟化酶(TH)阳性细胞数来评价PD小鼠模型建立情况。流式细胞仪检测各组小鼠脾脏悬液中Th17和MDSC的细胞百分率,并分析二者之间的相关性。结果:1、在PD患者外周血中,Th17和MDSC的细胞百分率较对照组明显升高,且二者升高程度呈正相关,结果均有显著差异。而Th17和MDSC与PD病情严重程度间未呈现相关性。2、MPTP建立的PD小鼠均出现不同程度的运动能力减退和运动行为异常。行为学实验中,相较于吉西他滨组和对照组,PD组和PD+吉西他滨组的小鼠在爬杆实验中调头与爬杆时间延长,悬挂实验中悬挂时间减少,强迫游泳实验中静止时间延长,结果均有显著差异。PD组和PD+吉西他滨组、吉西他滨组和对照组之间各行为学实验检查指标无明显差异。中脑黑质区TH细胞染色中,PD组和PD+吉西他滨组小鼠中脑黑质区TH细胞数量较吉西他滨组和对照组小鼠减少,结果具有显著差异。3、Th17细胞百分率在PD组小鼠脾脏悬液中较吉西他滨组和对照组升高,PD+吉西他滨组小鼠脾脏悬液中Th17细胞百分数较其余三组升高,结果均有显著差异。吉西他滨组和对照组之间无明显差异。4、MDSC细胞百分率在PD组小鼠脾脏悬液中较其他三组升高,在PD+吉西他滨组较吉西他滨组和对照组升高,结果均有显著差异。吉西他滨组和对照组之间无明显差别。5、在PD小鼠模型脾脏悬液中,各组Th17和MDSC未呈现相关性。结论:1、PD中存在神经免疫炎性反应,Th17与MDSC参与了该病理机制。2、在初诊PD患者外周血中,Th17与MDSC的表达较健康人明显升高。3、本研究中PD模型建立成功,且吉西他滨在PD模型中未对PD病理和症状产生明显影响。4、在PD小鼠模型的脾脏悬液中,Th17与MDSC的表达较正常小鼠明显升高。5、Th17和MDSC可能共同参与并促进了PD炎性反应的发生发展。6、在MPTP建立的亚急性PD小鼠模型中,MDSC可发挥其免疫调节作用抑制Th17扩增。
[Abstract]:Objective: to investigate the expression and correlation of helper T cell 17 (T17) and myeloid suppressor cell (MDSC) in PD mice with newly diagnosed Parkinson's disease (PD). To investigate the expression of Th17 and MDSC in the course of PD and to elucidate the mechanism of their action in PD. Method: 1. A total of 32 PD patients who were newly diagnosed and not treated with anti-PD were selected as PD group. The severity of the disease was evaluated by Hoehr-Yahr grading and unified Parkinson's disease rating scale. 32 healthy volunteers of the same age were selected as control group. Flow cytometry was used to detect the percentage of Th17 and MDSC in peripheral blood of the two groups, and Th17 was analyzed. The correlation between MDSC and severity of PD. 2. Forty C57BL / 6 mice were randomly divided into PD group and PD gemcitabine group. PD model was established with 1-methyl-4-phenyl-1-trihydropyridine-6-tetrahydropyridine (MPTP) in mice of gemcitabine group and control group (PD group and PD gemcitabine group). Mice in PD gemcitabine group and gemcitabine group were intraperitoneally injected with gemcitabine, while mice in control group were intraperitoneally injected with the same dose of normal saline. Hair and other manifestations, behavioral experiments (climbing test, hanging experiment, forced swimming experiment). Immunohistochemical detection of tyrosine hydroxylase in substantia nigra. To evaluate the establishment of PD mouse model, the percentage of Th17 and MDSC in spleen suspension of each group was detected by flow cytometry. Results the percentage of Th17 and MDSC cells in peripheral blood of PD patients was significantly higher than that of control group, and there was a positive correlation between them. There was no correlation between Th17 and MDSC and the severity of PD. PD mice established by MPTP showed different degrees of motor dysfunction and abnormal motor behavior. In behavioral experiment, compared with gemcitabine group and control group. PD group and PD gemcitabine group of mice in the climbing rod test in the rotation and climbing time prolonged, suspension test in the suspension time decreased, forced swimming test in the static time prolonged. Results there were significant differences between PD group and PD gemcitabine group. There was no significant difference in the behavioral parameters between the PD group and the control group. In the th cell staining of substantia nigra area of the mesencephalon, there was no significant difference between the two groups. The number of th cells in substantia nigra of PD group and PD gemcitabine group was significantly lower than that in gemcitabine group and control group. The percentage of Th17 cells in spleen suspension of PD group was higher than that of gemcitabine group and control group. The percentage of Th17 cells in spleen suspension of PD gemcitabine group was higher than that of other three groups. There was no significant difference between the gemcitabine group and the control group. The percentage of MDSC cells in the spleen suspension of PD group was higher than that of the other three groups. In PD gemcitabine group compared with gemcitabine group and control group, the results were significantly different. There was no significant difference between gemcitabine group and control group. There was no correlation between Th17 and MDSC in each group. Conclusion Th17 and MDSC are involved in the pathological mechanism of PD. Conclusion Th17 and MDSC are involved in the peripheral blood of patients with PD. The expression of Th17 and MDSC was significantly higher than that of healthy subjects. The PD model was successfully established in this study, and gemcitabine had no significant effect on pathology and symptoms of PD in PD model. The expression of Th17 and MDSC in spleen suspension of PD mice was significantly higher than that in normal mice. Th17 and MDSC may be involved in and promote the development of PD inflammatory response. 6, in the subacute PD mouse model established by MPTP. MDSC can exert its immunomodulatory function to inhibit Th17 amplification.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742.5

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