α-Naphthoflavone (αNF) is a prototype flavone, also known as a modulator of aryl hydrocarbon receptor (AhR). In the present study, we investigated the molecular mechanisms of αNF-induced cytotoxic effects in HT22 mouse hippocampal neuronal cells. αNF induced apoptotic cell death via activation of caspase-12 and -3 and increased expression of endoplasmic reticulum (ER) stress-associated proteins, including C/EBP homologous protein (CHOP). Inhibition of ER stress by treatment with the ER stress inhibitor, salubrinal, or by CHOP siRNA transfection reduced αNF-induced cell death. αNF activated mitogen-activated protein kinases (MAPKs), such as p38, JNK, and ERK, and inhibition of MAPKs reduced αNF-induced CHOP expression and cell death. αNF also induced accumulation of reactive oxygen species (ROS) and an antioxidant, N-acetylcysteine, reduced αNF-induced MAPK phosphorylation, CHOP expression, and cell death. Furthermore, αNF activated c-Src kinase, and inhibition of c-Src by a kinase inhibitor, SU6656, or siRNA transfection reduced αNF-induced ROS accumulation, MAPK activation, CHOP expression, and cell death. Inhibition of AhR by an AhR antagonist, CH223191, and siRNA transfection of AhR and AhR nuclear translocator reduced αNF-induced AhR-responsive luciferase activity, CHOP expression, and cell death. Finally, we found that inhibition of c-Src and MAPKs reduced αNF-induced transcriptional activity of AhR. Taken together, these findings suggest that αNF induces apoptosis through ER stress via c-Src-, ROS-, MAPKs-, and AhR-dependent pathways in HT22 cells.

中文翻译：

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α-萘黄酮通过内质网应激通过HT22海马神经元细胞中的c-Src-，ROS-，MAPKs-和芳基烃受体依赖性途径诱导细胞凋亡。

α-萘黄酮（αNF）是原型黄酮，也被称为芳烃受体（AhR）的调节剂。在本研究中，我们调查了HT22小鼠海马神经元细胞中αNF诱导的细胞毒性作用的分子机制。αNF通过激活caspase-12和-3并诱导内质网（ER）应激相关蛋白（包括C / EBP同源蛋白（CHOP））的表达诱导凋亡细胞死亡。通过用ER应激抑制剂，salubinal或CHOP siRNA转染治疗来抑制ER应激可减少αNF诱导的细胞死亡。αNF激活了丝裂原激活的蛋白激酶（MAPK），例如p38，JNK和ERK，而抑制MAPK则降低了αNF诱导的CHOP表达和细胞死亡。αNF还诱导了活性氧（ROS）和抗氧化剂N-乙酰半胱氨酸的积累，降低αNF诱导的MAPK磷酸化，CHOP表达和细胞死亡。此外，αNF激活c-Src激酶，并通过激酶抑制剂SU6656或siRNA转染抑制c-Src减少了αNF诱导的ROS积累，MAPK激活，CHOP表达和细胞死亡。AhR拮抗剂CH223191对AhR的抑制作用以及ahRNA和AhR核转运子的siRNA转染降低了αNF诱导的AhR反应性荧光素酶活性，CHOP表达和细胞死亡。最后，我们发现抑制c-Src和MAPKs可降低αNF诱导的AhR转录活性。综上所述，这些发现表明αNF通过HT22细胞中的c-Src-，ROS-，MAPKs-和AhR依赖性途径通过内质网应激诱导凋亡。激酶抑制剂SU6656或siRNA转染抑制c-Src可减少αNF诱导的ROS积累，MAPK活化，CHOP表达和细胞死亡。AhR拮抗剂CH223191对AhR的抑制作用以及ahRNA和AhR核转运子的siRNA转染降低了αNF诱导的AhR反应性荧光素酶活性，CHOP表达和细胞死亡。最后，我们发现抑制c-Src和MAPKs可降低αNF诱导的AhR转录活性。综上所述，这些发现表明αNF通过HT22细胞中的c-Src-，ROS-，MAPKs-和AhR依赖性途径通过内质网应激诱导凋亡。激酶抑制剂SU6656或siRNA转染抑制c-Src可减少αNF诱导的ROS积累，MAPK活化，CHOP表达和细胞死亡。AhR拮抗剂CH223191对AhR的抑制作用以及ahRNA和AhR核转运子的siRNA转染降低了αNF诱导的AhR反应性荧光素酶活性，CHOP表达和细胞死亡。最后，我们发现抑制c-Src和MAPKs降低了αNF诱导的AhR转录活性。综上所述，这些发现表明αNF通过HT22细胞中的c-Src-，ROS-，MAPKs-和AhR依赖性途径通过内质网应激诱导凋亡。siRNA转染AhR和AhR核易位子可降低αNF诱导的AhR响应荧光素酶活性，CHOP表达和细胞死亡。最后，我们发现抑制c-Src和MAPKs可降低αNF诱导的AhR转录活性。综上所述，这些发现表明αNF通过HT22细胞中的c-Src-，ROS-，MAPKs-和AhR依赖性途径通过内质网应激诱导凋亡。siRNA转染AhR和AhR核易位子可降低αNF诱导的AhR响应荧光素酶活性，CHOP表达和细胞死亡。最后，我们发现抑制c-Src和MAPKs可降低αNF诱导的AhR转录活性。综上所述，这些发现表明αNF通过HT22细胞中的c-Src-，ROS-，MAPKs-和AhR依赖性途径通过内质网应激诱导凋亡。