脂多糖对大鼠中脑原代培养细胞的神经损伤作用  

Injury effects of LPS on mesencephalic neuron-glia cultures of rats

作　　者：樊建鑫[1] 李治华[1] 任秀花[1] 张华[1] 周明付[1] 臧卫东[1] 

机构地区：[1]郑州大学基础医学院人体解剖学教研室,郑州450001

出　　处：《郑州大学学报（医学版）》2011年第1期38-42,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目311700531164

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。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.

关 键 词：多巴胺能神经元 脂多糖 白细胞介素6 离子钙结合蛋白1 大鼠 

分 类 号：R742.5[医药卫生—神经病学与精神病学]