【摘要】 目的:探讨脂多糖诱导帕金森病大鼠模型过程中肿瘤坏死因子α、白细胞介素1β、诱导型一氧化氮合酶等细胞毒性因子的作用。方法:实验于2003-07/2004-07在中山大学附属第一医院神经科实验室进行,取35只雄性SD大鼠随机分为4组:7,14,30d组,每组10只,对照组5只。对照组不做任何处理,3个不同时间点组立体定位注射脂多糖(20μg,质量浓度5g/L)入大鼠黑质。于注射后不同时间点观察大鼠行为学改变,并采用免疫组织化学及原位杂交等方法动态观察酪氨酸羟化酶神经元、肿瘤坏死因子α、白细胞介素1β、诱导型一氧化氮合酶等的表达。结果:29只动物进入结果分析。①行为学改变:脂多糖术后7d仅表现为轻度的旋转,每30min旋转(85±13)r,至14d时旋转次数增多,每30min旋转(121±17)r,30d时达高峰,每30min旋转(295±21)r。②7,14,30d组术侧黑质酪氨酸羟化酶阳性神经元数量较正常组明显下降(P<0.001),30d组下降达高峰;正常组仅有少量或无肿瘤坏死因子α、白细胞介素1β、诱导型一氧化氮合酶的表达,术后7d各阳性神经元表达较正常组显著增多(P<0.05),术后14d表达至高峰,术后30d较14d明显下降但仍高于正常对照组,其中诱导型一氧化氮合酶阳性神经元表达于术后30d仍维持较高水平。③各组均检测到酪氨酸羟化酶mRNA表达,7,14,30d组表达更多还原

【Abstract】 AIM: To study the role of tumor necrosis factor α , interleukin 1β and induced nitricoxide synthase in rats with Parkinson disease due to lipopolysaccharide.METHODS: The experiment was performed in the Laboratory of Department of Neurology, First Affiliated Hospital of Sun Yat sen University from July 2003 to July 2004.A total of 35 SD rats were divided randomly into 7, 14, 30 days groups(n=10) and control group(n=5). Rats at three different time points were injected with lipopolysaccharide(20 μ g, concentration >5 g/L). The behavior of rats was observed, and the expression of tyroxine hydroxylase neurons, tumor necrosis factor α , interleukin 1β and induced nitricoxide synthase was observed by using immunohistochemistry and in situ hybridization.RESULTS: A total of 29 rats were involved in the result analysis. ① The slight rotation was observed on the 7th day after operation [(85± 13)r per 30 minutes], the times increased on the 14th day [(121± 17)r per 30 minutes], and peaked on the 30th day [(295± 21)r per 30 minutes].② The positive neurons of tyroxine hydroxylase decreased significantly on the 7th, 14th and 30th days as compared with that in control group (P< 0.001), which reached to the lowest on the 30th day. The positive expression of tumor necrosis factor α , interleukin 1β and induced nitricoxide synthase increased obviously on the 7th day as compared with that in control group with little or no expression (P< 0.001), and increased to the peak on the 14th day; on the 30th day, it decreased significantly but was still higher than that in control group; the positive expression of induced nitricoxide synthase was still at a higher level on the 30th day. ③ The expression of tyroxine hydroxylase mRNA was detected in every group, and decreased on the 7th , 14th and 30th days as compared with that in control group (P < 0.01), reached to the lowest on 30th day(P< 0.001). The expression of tumor necrosis factor α , interleukin 1β and induced nitricoxide synthase mRNA was slight, which increased significantly on the 7th day after operation, and reached to the peak on the 14th day; the expression of tumor necrosis factor α , interleukin 1β decreased obviously on the 30th day following operation, but the expression of induced nitricoxide synthase mRNA still remained at a higher level.CONCLUSION: The positive expression of tumor necrosis factor α , interleukin 1β and induced nitricoxide synthase can be induced by lipopolysaccharide; the expressive peak is obviously earlier than the decreasing period of dopaminergic neuron, which indicates the cytotoxicity factors play a concerted role in dopaminergic neuronal degeneration, and accelerate the injury of neurons and the development of Parkinson disease.So it can block or relieve the vicious cycle to inhibit the development of immunity or inflammation.更多还原