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.
BACKGROUND
Da-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).


OBJECTIVE
To 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.


METHODS
Human 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.


RESULTS
Transfection 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.


CONCLUSIONS
These 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.
Department of Anatomy, School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing 100029, China; Center for Neurologic Diseases, Department of Neurology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA. Electronic address: zy_dalian@yahoo.com.
Yi
Y
Zhang
Department of Anatomy, School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing 100029, China; College of Special Education, Beijing Union University, Beijing 100075, China.
Xiao-Gang
XG
Gong
State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica & Neuroscience Center, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Zhen-Zhen
ZZ
Wang
Department of Anatomy, School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing 100029, China. Electronic address: hm.sun@yahoo.com.
Hong-Mei
HM
Sun
Department of Anatomy, School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
Zhen-Yu
ZY
Guo
Department of Anatomy, School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
Cong
C
Gai
Center for Scientific Research, School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
Jing-Hong
JH
Hu
Department of Anatomy, School of Preclinical Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
Ling
L
Ma
Beijing Key Lab for Immune-Mediated Inflammatory Diseases, Institute of Clinical Medical Science, China-Japan Friendship Hospital, Beijing 100029, China.
Ping
P
Li
State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica & Neuroscience Center, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China; College of Pharmacy, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China.
Nai-Hong
NH
Chen
D015655:1-Methyl-4-phenylpyridinium; D000255:Adenosine Triphosphate / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D042967:Electron Transport Complex I / Q000378:metabolism; D006801:Humans; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D009447:Neuroblastoma; D018696:Neuroprotective Agents / Q000494:pharmacology*; D000071617:Protein Deglycase DJ-1 / Q000235:genetics / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects
D004365:Drugs, Chinese Herbal; D018696:Neuroprotective Agents; C530014:dabuyin wan; D000255:Adenosine Triphosphate; D051057:Proto-Oncogene Proteins c-akt; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1; D042967:Electron Transport Complex I; D015655:1-Methyl-4-phenylpyridinium
Akt; DJ-1; Da-Bu-Yin-Wan; Mitochondrial dysfunction; Parkinson's disease; Phosphorylation