Protective effect of ginsenoside Rg1 on dopamine-induced apoptosis in PC12 cells.
OBJECTIVE
To explore the possible molecular mechanism of exogenous dopamine-induced apoptosis in PC12 cells and the protective effect of ginsenoside Rg1.


METHODS
Flow cytometric assay was used to quantify the apoptotic cells and measure the percentage of cells with positive Bcl-2 and Bax proteins. The morphology of apoptotic cells was evaluated by transmission electron microscope. DNA fragmentation was observed by gel electrophoresis. Caspase-3 activity was determined by fluorescent spectrofluorometer and the expressive bcl-2 and bax mRNA by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).


RESULTS
Dopamine 0.15, 0.30, 0.45, and 0.60 mmol/L induced PC12 cell apoptosis from 1.1 % +/- 0.4 % (control) to 41 % +/- 3 %, 46.4 % +/ -2.7 %, 53 % +/ -3 %, and 64.5 % +/- 2.7 %, respectively. After treatment with dopamine 0.45 mmol/L following pretreatment with Rg1 10 micromol/L for 24 h, the percentage of apoptotic cells and caspase-3 activity decreased from 53 % +/- 3 % and 683 +/- 8 (mean fluorescence intensity, MFI) to 1.9 % +/- 0.6 % and 325 +/- 5, and the percentage of cells with positive Bcl-2 protein increased from 14.3 % +/- 1.1 % to 25.9 % +/- 1.6 %, however, the percentage of cells with positive Bax protein decreased from 48 % +/- 3 % to 35 % +/- 3 %, compared with group treated with DA 0.45 mmol/L alone.


CONCLUSIONS
Ginsenoside Rg1 protected PC12 cells against apoptosis by inhibiting the activation of caspase-3 and regulating the ratio of Bcl-2 to Bax protein.
Chen
X C
XC
Fujian Institute of Geriatrics, Union Hospital, Fujian Medical Univisity, Fuzhou 350001, China. ZJCXCH@Public5.FZ.FJ.cn
Zhu
Y G
YG
Wang
X Z
XZ
Zhu
L A
LA
Huang
C
C
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D004298:Dopamine / Q000494:pharmacology*; D036145:Ginsenosides / Q000494:pharmacology*; D016716:PC12 Cells; D020011:Protective Agents / Q000494:pharmacology*; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D051028:bcl-2-Associated X Protein
D000975:Antioxidants; C494828:Bax protein, rat; D036145:Ginsenosides; D020011:Protective Agents; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; C035054:ginsenoside Rg1; D004298:Dopamine