阿特拉津对青春期大鼠黑质区多巴胺能神经元的损伤作用

doi:10.3969/j.issn.1004-616x.2017.04.002

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (4): 251-255.doi: 10.3969/j.issn.1004-616x.2017.04.002

阿特拉津对青春期大鼠黑质区多巴胺能神经元的损伤作用

马焜¹, 吴昊宇¹, 李鹏¹, 王丽², 李百祥¹

1. 哈尔滨医科大学公共卫生学院卫生毒理学教研室, 黑龙江哈尔滨 150081;
2. 哈尔滨商业大学校医院, 黑龙江哈尔滨 150028

收稿日期:2016-11-17 修回日期:2017-04-10 出版日期:2017-07-31 发布日期:2017-07-31
通讯作者: 李百祥,E-mail:lbxhrb@163.com E-mail:lbxhrb@163.com
作者简介:马焜,E-mail:kris_mamm@hotmail.com。
基金资助:
国家自然科学基金资助项目（81273109）

Induction of dopaminergic neuron degeneration by atrazine in substantia nigra in adolescent rats

MA Kun¹, WU Haoyu¹, LI Peng¹, WANG Li², LI Baixiang¹

1. Department of Toxicology, Public Health College of Harbin Medical University, Harbin 150081;
2. School Hospital, Harbin University of Commerce, Harbin 150028, Heilongjiang, China

Received:2016-11-17 Revised:2017-04-10 Online:2017-07-31 Published:2017-07-31

摘要/Abstract

摘要：

目的：探讨阿特拉津对青春期大鼠黑质区多巴胺能神经元的损伤作用。方法：将4周龄的Wistar雄性大鼠40只随机分为4组，分别为对照组（1%甲基纤维素）、阿特拉津低（50 mg/kg）、中（100 mg/kg）和高（200 mg/kg）剂量组，每组10只，连续灌胃45 d后处死。收集中脑黑质部分，采用免疫组织化学法（SP法）检测神经元细胞中酪氨酸羟化酶（TH）的变化，并采用实时定量PCR法（qPCR）和Western blot检测多巴胺能神经元损伤相关基因mRNA和蛋白的表达情况。结果：免疫组化结果显示，200 mg/kg ATR组中TH阳性细胞数较对照组显著减少，差异具有统计学意义（P < 0.01）。qPCR结果显示，与对照组相比，100和200 mg/kg ATR染毒组TH mRNA和Bcl-2 mRNA的表达量显著降低；200 mg/kg ATR组p53 mRNA和Caspase-9 mRNA的表达量显著增加，差异具有统计学意义（P < 0.05）。Western blot结果显示，与对照组相比，100和200 mg/kg ATR染毒组TH和Bcl-2的蛋白表达量显著降低；100和200 mg/kgATR组p53蛋白表达量显著增加；50、100和200 mg/kg ATR组Caspase-9蛋白表达量均显著增加，差异均具有统计学意义（P均 < 0.05）。结论：阿特拉津可以影响大鼠黑质区多巴胺能神经元损伤相关基因mRNA和蛋白的表达。

关键词: 阿特拉津, 多巴胺能神经元, 免疫组织化学, 酪氨酸脱氢酶

Abstract:

OBJECTIVE: To study effects of atrazine (ATR) on neuronal degeneration in adolescent rats. METHODS: 4 weeks old Wistar male rats were randomly subdivided into 4 groups:control and ATR test groups. All rats were treated with ATR:50,100 and 200 mg/kg body weight per day (five times per week) for 45 days by oral gavage. They were sacrificed at 12 h after the last ATR injection. The entire midbrain was collected and stored frozen at -80℃. The presence of tyrosine hydroxylase(TH) was detected by immunohistochemistry SP in dopaminergic neurons. TH expression in mRMA and protein was measured using real-time PCR and Western blot. RESULTS: Immunohistochemical results showed that,compared with the control group,the number of TH positive cells in the 200 mg/kg ATR group was significantly decreased (P < 0.01). In addition,qPCR results show that expression of TH mRNA and Bcl-2 mRNA was significantly decreased in the 100 and 200 mg/kg ATR groups,and expression of p53 mRNA and Caspase-9 mRNA was significantly increased in the 200 mg/kg ATR group,the difference was statistically significant (P < 0.05). Western blot results show that expression of TH and Bcl-2 protein was significantly decreased in the 100 and 200 mg/kg ATR groups compared with the control group. Expression of p53 protein in the ATR group was significantly increased at 100 and 200 mg/kg,and Caspase-9 protein in he 200 mg/kg ATR group was significantly increased,the difference was statistically significant (P < 0.05). CONCLUSION: Atrazine caused damage to the dopaminergic neurons in the substantia nigra of rats and activated the apoptotic pathway which led to the dopaminergic neuronal death.

Key words: atrazine, dopaminergic neurons, immunohistochemistry, tyrosine hydroxylase

中图分类号:

R741

马焜, 吴昊宇, 李鹏, 王丽, 李百祥. 阿特拉津对青春期大鼠黑质区多巴胺能神经元的损伤作用[J]. 癌变·畸变·突变, 2017, 29(4): 251-255.

MA Kun, WU Haoyu, LI Peng, WANG Li, LI Baixiang. Induction of dopaminergic neuron degeneration by atrazine in substantia nigra in adolescent rats[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(4): 251-255.

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阿特拉津对青春期大鼠黑质区多巴胺能神经元的损伤作用

Induction of dopaminergic neuron degeneration by atrazine in substantia nigra in adolescent rats

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