Lin Chia-Yuan, Fu Ru-Huei, Chou Ruey-Hwang, Chen Jing-Hsien, Wu Chi-Rei, Chang Shu-Wei, Tsai Chia-Wen
Graduate Institute of Biomedical Sciences and Center for Molecular Medicine, China Medical University, Taichung, Taiwan.
Graduate Institute of Biomedical Sciences, China Medical University, Taichung, Taiwan; Translational Medicine Research Center, China Medical University Hospital, Taichung, Taiwan.
Food Chem Toxicol. 2017 May;103:194-202. doi: 10.1016/j.fct.2017.03.020. Epub 2017 Mar 10.
Pi class of glutathione S-transferase (GST) is known to suppress c-Jun N-terminal kinase (JNK)-related apoptosis through protein-protein interactions. Moreover, signaling by PKA/cAMP response element binding protein (CREB) is necessary for GSTP up-regulation. This study explored whether carnosic acid (CA) from rosemary prevents 6-hydroxydopamine (6-OHDA)-induced neurotoxicity by inhibition of JNK through GSTP via PKA/CREB signaling. Results indicated that the GSTP protein was increased in SH-SY5Y cells treated with CA for 18 and 24 h. However, CA had no significant effect on alpha or mu class of GST. Treatment of CA increased the induction of p-PKAα, nuclear p-CREB, and CRE-DNA binding activity. These effects of CA were attenuated in cells pretreated with the PKA inhibitor H89. CA pretreatment suppressed 6-OHDA-induced apoptosis by inhibition of JNK phosphorylation, poly(ADP)-ribose polymerase cleavage, and nuclear condensation. Pretreatment with H89 and GSTP siRNA attenuated the ability of CA to reverse 6-OHDA-induced apoptosis. By use of immunoprecipitation with JNK antibody to examine the interaction of GSTP-JNK with CA, we showed that CA pretreatment increased the immunoprecipitation of GSTP after 6-OHDA treatment, which suggests that CA promoted the interaction between GSTP and JNK.
CA prevents 6-OHDA-induced apoptosis via inhibition of JNK by GSTP through the PKA/CREB pathway.
已知谷胱甘肽S-转移酶（GST）的Pi类通过蛋白质-蛋白质相互作用抑制c-Jun氨基末端激酶（JNK）相关的细胞凋亡。此外，蛋白激酶A/环磷酸腺苷反应元件结合蛋白（PKA/CREB）信号传导对于GSTP的上调是必需的。本研究探讨了迷迭香中的鼠尾草酸（CA）是否通过PKA/CREB信号传导途径经由GSTP抑制JNK来预防6-羟基多巴胺（6-OHDA）诱导的神经毒性。结果表明，用CA处理18和24小时的SH-SY5Y细胞中GSTP蛋白增加。然而，CA对GST的α或μ类没有显著影响。CA处理增加了p-PKAα、核p-CREB的诱导以及CRE-DNA结合活性。CA的这些作用在用PKA抑制剂H89预处理的细胞中减弱。CA预处理通过抑制JNK磷酸化、聚（ADP-核糖）聚合酶裂解和核浓缩来抑制6-OHDA诱导的细胞凋亡。用H89和GSTP siRNA预处理减弱了CA逆转6-OHDA诱导的细胞凋亡的能力。通过使用JNK抗体免疫沉淀来检测GSTP-JNK与CA的相互作用，我们发现CA预处理增加了6-OHDA处理后GSTP的免疫沉淀，这表明CA促进了GSTP与JNK之间的相互作用。
CA通过PKA/CREB途径经由GSTP抑制JNK来预防6-OHDA诱导的细胞凋亡。
