【摘要】 目的:观察脂多糖(LPS)对大鼠中脑原代混合细胞多巴胺能神经元的损伤作用。方法:取孕14d的SD大鼠胚胎中脑原代培养。①将细胞分为正常对照组和不同剂量LPS(0.01、0.10、1.00和10.00mg/L)组,体外培养7d后,实验组加入不同浓度的LPS,持续作用72h后,经酪氨酸羟化酶(TH)免疫细胞化学染色,计数TH阳性细胞数目并观察形态变化。②将细胞分为正常对照组和0.1mg/L的LPS组。体外培养7d,实验组加入终浓度为0.1mg/L的LPS,分别作用8、24和72h,收集细胞上清液,用ELISA检测白细胞介素6(IL-6)的含量;分别对作用8、24和72h的原代细胞进行TH和离子钙结合蛋白1(Iba1)的免疫细胞化学染色,计数多巴胺能神经元和激活的小胶质细胞的数量。结果:TH阳性细胞减少,大部分细胞表现为突起数目的减少,突起变短并有串珠样改变。0.1mg/LLPS分别作用8、24和72h的给药组与对照组相比,IL-6的含量均有显著增加(t分别为15.787、16.712和15.910,P均<0.001)。激活的Iba1染色阳性神经元的数量分别为(40.0±6.1)、(76.0±3.4)和(69.0±5.2)(F=387.820,P<0.001)。在24和72h的LPS给药组中,TH染色阳性神经元的数量分别为(18.0±5.3)和(10.0±3.2)(F=94.840,P<0.001)。结论:LPS通过激活小胶质细胞释放IL-6发挥对多巴胺能神经元的损伤作用。更多还原

【Abstract】 Aim:To investigate the injury effects of dopaminergic neurons induced by LPS in rat mesencephalic neuron-glia cultures.Methods:Rat mesencephalic neuron-glia cultures were obtained from timed pregnant SD-rats on embryonic 14 d.Ⅰ:The mesencephalic neuron-glia cultures were categorized into five groups:control and LPS(0.01,0.10,1.00,and 10.00 mg/L)group.On the 7th day of culture,different concentrations of LPS were incubated for 72 h.Dopaminergic neurons were identified and counted by using tyrosine hydroxylase(TH)immunochemistry.Ⅱ:The mesencephalic neuron-glia cultures were categorized into two groups:control and LPS group(0.1 mg/L).On the 7th day of culture,0.1 mg/L LPS was incubated for different times(8,24,and 72 h).After administration supernatants were collected,IL-6 levels were detected by using ELISA.Dopaminergic neurons and actived microglias were identified and counted by using TH and ionized calcium binding adaptor molecule 1(Iba1)immunochemistry.Results:The number of TH positive cells reduced significantly with shorter cell processes showing bead-like.The IL-6 levels in 0.1 mg/L LPS-treated groups of 8,24 and 72 h after administration,increased significantly(t was 15.787,16.712 and 15.910,P<0.001).The number of actived microglias were(40.0±6.1),(76.0±3.4),and(69.0±5.2),respectively(F=387.820,P<0.001).The number of dopaminergic neurons were(18.0±5.3)and(10.0±3.2)in 0.1 mg/L LPS-treated groups of 24 and 72 h after administration,respectively(F=94.840,P<0.001).Conclusion:Microglias may play an important role in injury effects of dopaminergic neurons,activated by LPS to release IL-6.更多还原