Study on the effects of stress hormone and endocrine disrupter on an organism using the technology in neuroscience

利用神经科学技术研究应激激素和内分泌干扰物对生物体的影响

• 批准号: 15590504
• 负责人: HOSOKAWA Toshiyuki
• 金额: $ 1.86万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590504/
• 关键词: Stress; Endocrine disrupter; Cell Culture; Apoptosis; PC12; Hippocampus; ホルモン; シナプス

The object of this project was to study the effect of the stress hormones and the endocrine disrupters to the microscopic structure of nerve tissue.New imaging method was applied to the living hippocampal slices in the chapter 4. It showed spatiotemporal patterns of the long-term potentiation (LTP). It will be the strong method to study the brain under the effect of such endocrine disrupters.The effect of nonylphenol on apoptosis in PC12 cells was studied. We investigated effect of nonylphenol on apoptosis induced by serum deprivation in PC12 cells using TUNEL and DNA fragmentation assays. In addition, changes in contents of proapoptotic factors, Bad and Bax, and antiapoptotic factor, Bcl-2, and enzyme activity of caspase-3 were studied. Below 100 ng/ml of nonylphenol increased TUNEL signals, DNA fragmentation and content of proapoptoticfactor, Bad as compared to those by serum deprivation without nonylphenol. Furthermore, addition of nonylphenol enhanced caspase-3 activity and Z-VAD, caspase-3 inhibitor, diminished such effect. These results indicated that below 100 ng/ml of nonylphenol enhanced apoptosis induced by serum deprivation via caspase-3 activation in PC12 cell.The changes of apoptosis of cultured cells are investigated under the existence of small quantities of cortisol. Moreover the changes of the relational factors of the apoptosis were also studied using PC12 or CHO cells. The apoptosis was induced with 6OHDA or without FBS. On set of the apoptosis was measured by TUNEL method, The RT-PCR method was used to measure the m-RNA of apoptosis related factors Bax and Bcl-2. 100 nM and 10μM of cortisol inhibits the apoptosis. The signals of Bax showed significant increase compared with control (P<0.1).

本课题的目的是研究应激激素和内分泌干扰物对神经组织微观结构的影响。第四章将新的成像方法应用于活体海马片，显示了长时程增强(LTP)的时空模式。这将是研究这类内分泌干扰物作用下大脑的有力手段。本文研究了壬基酚对PC12细胞凋亡的影响。采用原位末端标记法和DNA片段化分析方法，研究了壬基酚对血清剥夺诱导PC12细胞凋亡的影响。此外，还观察了促凋亡因子Bad、Bax和抗凋亡因子Bcl2含量及caspase 3酶活性的变化。低于100 ng/ml的壬基酚可增加TUNEL信号、DNA片段化和促凋亡因子的含量，与无血清剥夺组比较，差异有统计学意义。此外，加入壬基酚可增强caspase-3的活性，而caspase-3的抑制剂Z-VAD可减弱这种作用。这些结果表明，低于100 ng/ml的壬基酚通过激活caspase-3来增强血清剥夺诱导的PC12细胞的凋亡。此外，还利用PC12和CHO细胞研究了细胞凋亡相关因素的变化。分别用6OHDA或不加FBS诱导细胞凋亡。原位末端标记法检测细胞凋亡率，RT-PCR法检测细胞凋亡相关因子Bax、Bcl2的m-RNA表达。100 nM和10μM的皮质醇对细胞凋亡有抑制作用。与对照组相比，Bax蛋白表达明显增强(P&lt；0.1)。

项目成果

T.Hosokawa et al.: "Imaging Spatiotemporal Patterns of Long-Term Potentiation in Mouse Hippocampus"Philos Trans R Soc Lond B Biol Sci.. 358(1432). 689-693 (2003)

T.Hosokawa 等人：“小鼠海马长期增强的时空模式成像”Philos Trans R Soc Lond B Biol Sci. 358(1432)。

Decrease in cytosolic phospholipase A2alpha mRNA levels by reactive oxygen species via MAP kinase pathways in PC12 cells : effects of dopaminergic neurotoxins

活性氧通过 PC12 细胞中 MAP 激酶途径降低胞质磷脂酶 A2α mRNA 水平：多巴胺能神经毒素的影响

• 发表时间: 2005
• 期刊: Cellular Signalling 17
• 作者: Akiyama;N
• 通讯作者: N

Nonylphenol enhances apoptosis induced by serum deprivation in PC12 cells.

• DOI: 10.1016/j.lfs.2003.09.066
• 发表时间: 2004-03
• 期刊: Life sciences
• 影响因子: 6.1
• 作者: M. Aoki;M. Kurasaki;T. Saito;Sayaka Seki;T. Hosokawa;Yasumitsu Takahashi;H. Fujita;T. Iwakuma

Long-term potentiation in the optic tectum of rainbow trout

虹鳟鱼视顶盖的长时程增强

• 发表时间: 2004
• 期刊: Neuroscience Letters 370巻
• 作者: T.Fujita;R.Katoh;A.H.Arieta;H.Yokoi;T.Arai;花田 智之;荻原克益;荻原克益;Masae Kinoshita;Masae Kinoshita
• 通讯作者: Masae Kinoshita

Immunohistochemical study of macrophage migration inhibitory factor in rat liver fibrosis induced by thioacetamide.

硫代乙酰胺诱导大鼠肝纤维化巨噬细胞迁移抑制因子的免疫组织化学研究

• 发表时间: 2003
• 期刊: European Journal of Histochemistry 47
• 作者: Y.Hori;et al.
• 通讯作者: et al.