Protective effects of DJ-1 medicated Akt phosphorylation on mitochondrial function are promoted by Da-Bu-Yin-Wan in 1-methyl-4-phenylpyridinium-treated human neuroblastoma SH-SY5Y cells
Protective effects of DJ-1 medicated Akt phosphorylation on mitochondrial function are promoted by Da-Bu-Yin-Wan in 1-methyl-4-phenylpyridinium-treated human neuroblastoma SH-SY5Y cells
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大补饮丸在 1-甲基-4-苯基吡啶鎓处理的人神经母细胞瘤 SH-SY5Y 细胞中促进 DJ-1 药物 Akt 磷酸化对线粒体功能的保护作用
DOI：
10.1016/j.jep.2016.04.029
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发表时间：
2016
影响因子：
5.4
通讯作者：
Nai-Hong Chen
中科院分区：
文献类型：
--
作者：
Yi Zhang;Xiao-Gang Gong;Zhen-Zhen Wang;Hong-Mei Sun;Zhen-Yu Guo;Cong Gai;Jing-Hong Hu;Ling Ma;Ping Li;Nai-Hong Chen
民族药理学相关性大补阴丸（DBYW）是一个历史悠久的传统中药配方，最初定义于600多年前。本研究从DJ-1和Akt信号通路的角度探讨了DBYW对线粒体功能的影响，旨在探讨DBYW对线粒体功能的潜在机制。转染的细胞用1-甲基-4-苯基吡啶鎓（MPP+）（一种PD相关的线粒体复合物I抑制剂）在不存在和存在DBYW的情况下处理。通过细胞计数试剂盒-8测定法评估细胞活力。Western blotting检测DJ-1和Akt信号通路蛋白的表达。通过共聚焦荧光显微镜评估线粒体质量。结果转染pcDNA 3-Flag-DJ-1可降低MPP+诱导的细胞毒性，并过表达DJ-1。在DJ-1过表达的细胞中，线粒体质量增加，线粒体复合物I活性提高，细胞ATP含量增加。此外，DJ-1的过表达增强了苏氨酸308和丝氨酸473上的Akt磷酸化。结论DJ-1可能通过抑制MPP+诱导的SH-SY 5 Y细胞Akt磷酸化而保护线粒体功能。此外，DBYW增强DJ-1介导的Akt磷酸化对线粒体功能的保护作用。
Ethnopharmacological relevanceDa-Bu-Yin-Wan (DBYW), a historically traditional Chinese medicine formula, was originally defined over 600 years ago. In recent decades, DBYW was clinically employed to treat Parkinson's disease (PD).Aim of the studyTo explore the underlying mechanism of DBYW on mitochondrial function, we investigated the effect of DBYW on mitochondrial function from the perspectives of DJ-1 and Akt signaling.Materials and methodsHuman derived neuroblastoma SH-SY5Y cells were transiently transfected with the plasmid pcDNA3-Flag-DJ-1 aimed to overexpress the DJ-1 protein. Transfected cells were treated with 1-methyl-4-phenylpyridinium (MPP+), a PD-related mitochondrial complex I inhibitor, in the absence and presence of DBYW. The cell viability was assessed by Cell Counting Kit-8 assay. The protein expressions of DJ-1 and Akt signaling were examined by western blotting. The mitochondrial mass was evaluated by confocal fluorescence microscopy. The mitochondrial complex I activity and cellular ATP content were measured by commercial kits.ResultsTransfection with pcDNA3-Flag-DJ-1 decreased the MPP+-induced toxicity and overexpressed the DJ-1. In DJ-1 overexpressed cells, the mitochondrial mass was raised, mitochondrial complex I activity was improved, and cellular ATP content was increased. In addition, overexpression of DJ-1 augmented the Akt phosphorylation on threonine 308 and serine 473. Moreover, DBYW promoted the above effects in DJ-1 expressed cells.ConclusionsThese data suggest that DJ-1 protects the mitochondrial function by medicating Akt phosphorylation in MPP+-treated SH-SY5Y cells. Moreover, DBYW enhances the protective effect of DJ-1 medicated Akt phosphorylation on mitochondrial function.