Abstract | OBJECTIVE: METHODS: The pheochromocytoma (PC12) cell line was employed in the present study. Cells were treated with 2 mumol/L PKCdelta inhibitor Rottlerin, 10 nmol/L protein kinase C delta subtype (PKCdelta) inhibitor bisindolylmaleimide I, or 5 nmol/L Gö6976 that could specifically inhibit the calcium-dependent PKCdelta isoforms, respectively. PKC activator phorbol-12-myristate-13-acetate (PMA, 100 nmol/L) was also used in this study. All these agents were added to the medium before cells were incubated with 6-OHDA. Cells with no treatment served as control. The cytotoxicity of 6-OHDA was determined by methyl thiazolyl tetrazolium (MTT) reduction assay and PKCdelta phosphorylation levels in various groups were measured by western blotting. RESULTS:
Bisindolylmaleimide I and Gö6976 exerted no significant attenuation on the cytotoxicity of 6-OHDA, nor any effects on PKCdelta phosphorylation in PC12 cells. However, Rottlerin could inhibit the phosphorylation of PKCdelta and attenuate 6-OHDA-induced cell death, and the cell viability was raised to (69.6+/-2.63)% of that in control group (P<0.05). In contrast, PMA induced a significant increase in PKCdelta phosphorylation and also strengthened the cytotoxic effects of 6-OHDA. The cell viability of PMA-treated PC12 cells decreased to (49.8+/-5.06)% of that in control group (P<0.001). CONCLUSION:
Rottlerin can protect PC12 cells from cytotoxicity of 6-OHDA probably by inhibiting PKCdelta phosphorylation. The results suggest that PKCdelta may be a key regulator of neuron loss in Parkinson's disease.
|
Authors | Ying Fan, Yan-Qiao Zhang, Dian-Jun Sun, Yi-Na Zhang, Xiao-Wei Wu, Jing Li |
Journal | Neuroscience bulletin
(Neurosci Bull)
Vol. 25
Issue 4
Pg. 187-95
(Aug 2009)
ISSN: 1995-8218 [Electronic] Singapore |
PMID | 19633700
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
|
Chemical References |
- Acetophenones
- Adrenergic Agents
- Benzopyrans
- Carbazoles
- Enzyme Inhibitors
- Indoles
- Maleimides
- Phorbol Esters
- Go 6976
- 12-O-retinoylphorbol-13-acetate
- Oxidopamine
- rottlerin
- Protein Kinase C
- bisindolylmaleimide I
- Dopamine
|
Topics |
- Acetophenones
(pharmacology)
- Adrenergic Agents
(toxicity)
- Animals
- Benzopyrans
(pharmacology)
- Carbazoles
(pharmacology)
- Cell Survival
(drug effects)
- Dopamine
(metabolism)
- Dose-Response Relationship, Drug
- Drug Interactions
- Enzyme Activation
(drug effects)
- Enzyme Inhibitors
(pharmacology)
- Gene Expression Regulation, Enzymologic
(drug effects)
- Indoles
(pharmacology)
- Maleimides
(pharmacology)
- Oxidopamine
(toxicity)
- PC12 Cells
- Phorbol Esters
(pharmacology)
- Phosphorylation
(drug effects)
- Protein Kinase C
(metabolism)
- Rats
- Time Factors
|