Tian T, Zhang B Y, Wang K D, Zhang B F, Huang M
Department of Occupational Health and Environmental Hygiene, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2020 Nov 20;38(11):801-808. doi: 10.3760/cma.j.cn121094-20200121-00036.
To investigate the protective effect of taurine (Tau) on hippocampus, substantia nigra neurons and microglia in paraquat (PQ) -induced Pakinson's disease-like mice. In April 2019, the specific pathogen free (SPF) C57BL/6 mice (=36) were randomly divided into control group (NaCl) , Tau control group (150 mg/kg) , PQ exposure group (10 mg/kg PQ group, 15 mg/kg PQ group) , Tau intervention group (Tau+10 mg/kg PQ group, Tau+15 mg/kg PQ group) , respectively. Tau was used in 1 h before PQ administration for consecutive 6 weeks (twice per week) . General and neurobehavioral tests (Traction test, Open field test, Forced Swimming test, Tail suspension test, High plus maze and Object recognition test) were performed to test motor and cognitive function. After neuroethology detection, mice were euthanized and brains were collected. Nissl staining was used to detect the changes of the number and morphology of Nissl bodies in hippocampus and substantia nigra neurons of mice. Immunohistochemistry (IHC) was used to test the levels of neuron marker neuronal nuclei antigen (NeuN) , substantia nigra dopaminergic neuron marker tyrosine hydroxylase (TH) , α-synuclein (α-syn) , microglia markers ionized calcium bindingadaptor molecule-1 (Iba-1) , inducible nitric oxide synthase (iNOS) and interleukin-1β (IL-1β) in mice substantia nigra. The coexpression of Iba-1 and TH double-labeling, α-syn and TH double-labeling in mice substantia nigra were measured by immunofluorescence double staining. General behavioral changes such as slow reaction and reduced action occurred in mice of PQ group. Compared with the control group, the scores of Traction test, and the time ratio of new object recognition in the PQ group decreased (<0.05) , the fixed time of Swimming test and Tail suspension test increased (<0.05) , the horizontal crawl number and vertical times of Open field test and the ratio of open arm residence time of High plus maze in the 15 mg/kg PQ group decreased (<0.05) . Compared with the PQ group, the same dose of Tau+PQ group showed increased scores in Traction test (<0.05) and decreased fixed time of Swimming test and Tail suspension test (<0.05) . Compared with the 15 mg/kg PQ group, the horizontal crawl number of Open field test and the time ratio of new object recognition increased in the Tau+15 mg/kg PQ group (<0.05) . Compared with the control group, the PQ group showed a decrease in the number of Nissl body in the hippocampus and substantia nigra (<0.05) , a decrease in the number of NeuN and TH positive cells in the substantia nigra (<0.05) , with a large number of α-syn deposition, Iba-1 activation of microglia cells, and an increase in the expression of inflammatory factors (IL-1β, iNOS) in the hippocampus and substantia nigra (<0.05) . Compared with the PQ group, the same dose of Tau+PQ group showed the number of Nissl in the hippocampus and substantia nigra was significantly increased (<0.05) , the number of NeuN and TH positive cells in the substantia nigra was significantly increased (<0.05) , the expression levels of α-syn, Iba-1 and inflammatory factors (IL-1β, iNOS) in the substantia nigra were significantly decreased (<0.05) . Tau could protect PQ-induced degeneration of substantia nigra dopaminergic neurons and hippocampal neuron loss by inhibiting the activation of microglia cells and release of inflammatory factors, and effectively improve the neurobehavioral and brain histopathological changes of PQ-induced PD-like mice.
探讨牛磺酸(Tau)对百草枯(PQ)诱导的帕金森病样小鼠海马、黑质神经元及小胶质细胞的保护作用。2019年4月,将36只无特定病原体(SPF)的C57BL/6小鼠随机分为对照组(NaCl)、Tau对照组(150 mg/kg)、PQ暴露组(10 mg/kg PQ组、15 mg/kg PQ组)、Tau干预组(Tau + 10 mg/kg PQ组、Tau + 15 mg/kg PQ组)。在给予PQ前1小时使用Tau,连续6周(每周两次)。进行一般和神经行为测试(牵引试验、旷场试验、强迫游泳试验、悬尾试验、高架十字迷宫和物体识别试验)以测试运动和认知功能。神经行为学检测后,对小鼠实施安乐死并采集大脑。采用尼氏染色检测小鼠海马和黑质神经元中尼氏体数量和形态的变化。采用免疫组织化学(IHC)检测小鼠黑质中神经元标志物神经元细胞核抗原(NeuN)、黑质多巴胺能神经元标志物酪氨酸羟化酶(TH)、α-突触核蛋白(α-syn)、小胶质细胞标志物离子钙结合衔接分子-1(Iba-1)、诱导型一氧化氮合酶(iNOS)和白细胞介素-1β(IL-1β)的水平。通过免疫荧光双染色检测小鼠黑质中Iba-1和TH双标、α-syn和TH双标的共表达。PQ组小鼠出现反应迟钝、活动减少等一般行为变化。与对照组相比,PQ组牵引试验得分及新物体识别时间比例降低(<0.05),游泳试验和悬尾试验的固定时间增加(<0.05),15 mg/kg PQ组旷场试验的水平爬行次数和垂直次数以及高架十字迷宫开放臂停留时间比例降低(<0.05)。与PQ组相比,相同剂量的Tau + PQ组牵引试验得分增加(<0.05),游泳试验和悬尾试验的固定时间降低(<0.05)。与15 mg/kg PQ组相比,Tau + 15 mg/kg PQ组旷场试验的水平爬行次数和新物体识别时间比例增加(<0.05)。与对照组相比,PQ组海马和黑质中尼氏体数量减少(<0.05),黑质中NeuN和TH阳性细胞数量减少(<0.05),有大量α-syn沉积,小胶质细胞Iba-1激活,海马和黑质中炎症因子(IL-1β、iNOS)表达增加(<0.05)。与PQ组相比,相同剂量的Tau + PQ组海马和黑质中尼氏体数量显著增加(<0.05),黑质中NeuN和TH阳性细胞数量显著增加(<0.05),黑质中α-syn、Iba-1和炎症因子(IL-1β、iNOS)的表达水平显著降低(<0.05)。牛磺酸可通过抑制小胶质细胞激活和炎症因子释放,保护PQ诱导的黑质多巴胺能神经元变性和海马神经元丢失,并有效改善PQ诱导的帕金森病样小鼠的神经行为和脑组织病理学变化。