[Effect of chlorpyrifos on autophagy-related protein expression in hippocampal neurons of SD rats]

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2021 Sep 20;39(9):648-652. doi: 10.3760/cma.j.cn121094-20191120-00535.
[Article in Chinese]

Abstract

Objective: To analyze the effect of chlorpyrifos on the expression of autophagy related proteins in rat hippocampal neurons, and to explore the role of autophagy in central nerve injury caused by acute chlorpyrifos poisoning. Methods: In October 2018, 35 male clean grade SD rats were randomly divided into 7 groups according to the observation time point, namely 0.5 d, 1 d, 2 d, 3 d, 5 d and 7 d groups and the control group, with 5 rats in each group. Each observation group was given 81.5 mg/kg chlorpyrifos by gavage, and the control group was given olive oil by gavage. The general conditions and poisoning symptoms of rats were observed continuously after exposure. The expressions of autophagy related proteins Beclin1, P62/SQSTM1 and LC3 in hippocampus were detected by Western blot. The cell morphology and LC3 expression in brain were observed by immunohistochemical staining. Results: Western blot results showed that compared with the control group, the expression of Beclin1 protein in hippocampal neurons of rats in the 1 d, 2 d, and 3 d groups increased, while the expression of P62/SQSTM1 protein in the 0.5 d, 1 d, and 2 d groups decreased, and the expression of LC3 protein was decreased in the 2 d group, and the differences were statistically significant (P<0.05) . The results of immunohistochemistry showed that the hippocampal neurons of rats in the 5 d group were arranged disorderly, and some nuclei contours disappeared, especially in the 7 d group. The LC3 protein was expressed in the cytoplasm, and the expression level gradually increased, reaching a peak on the second day. Conclusion: The early activation of autophagy in rats with acute chlorpyrifos poisoning may be involved in chlorpyrifos induced hippocampal neuronal injury.

目的: 分析毒死蜱对大鼠海马神经元自噬相关蛋白表达的影响,探讨自噬在急性毒死蜱中毒中枢神经损伤中的作用。 方法: 于2018年10月,将35只雄性清洁级SD大鼠根据观察时间点随机分成7组,即0.5 d、1 d、2 d、3 d、5 d、7 d组和对照组,每组5只。各观察组采用81.5 mg/kg毒死蜱一次性灌胃处理,对照组给予橄榄油灌胃。连续观察大鼠一般情况和中毒症状,蛋白质免疫印迹(Western blot)法检测海马组织中自噬相关蛋白Beclin1、P62/SQSTM1、LC3的表达,免疫组织化学染色观察脑组织细胞形态和LC3表达。 结果: Western blot结果显示,与对照组比较,1 d、2 d、3 d组大鼠海马神经元Beclin1蛋白表达升高,0.5 d、1 d、2 d组P62/SQSTM1蛋白表达降低,2 d组大鼠海马神经元LC3蛋白表达升高,差异均有统计学意义(P<0.05)。免疫组织化学结果显示,5 d组大鼠海马神经元排列紊乱,部分细胞核轮廓消失,7 d组尤其明显,LC3蛋白呈胞质表达,表达量逐渐增高,第2天达高峰。 结论: 急性毒死蜱中毒大鼠自噬的早期激活可能参与了毒死蜱诱导的海马神经元损伤。.

Keywords: Autophagy; Beclin1; Chlorpyrifos; Hippocampus; LC3; Neurons; P62/SQSTM1; Rats.

MeSH terms

  • Animals
  • Autophagy-Related Proteins
  • Chlorpyrifos* / toxicity
  • Hippocampus
  • Male
  • Neurons
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Autophagy-Related Proteins
  • Chlorpyrifos