Limits...
The protective role of (-)-epigallocatechin-3-gallate in thrombin-induced neuronal cell apoptosis and JNK-MAPK activation.

He Q, Bao L, Zimering J, Zan K, Zhang Z, Shi H, Zu J, Yang X, Hua F, Ye X, Cui G - Neuroreport (2015)

Bottom Line: Flow cytometric analysis and western blotting demonstrated that thrombin-induced neuron degeneration occurs through apoptosis.These data suggest that EGCG may have protective effects against thrombin-induced neuroapoptosis by inhibiting the activation of JNK, leading to caspase 3 cleavage.EGCG is a novel candidate neuroprotective agent against intracerebral hemorrhage-induced neurotoxicity.

View Article: PubMed Central - PubMed

Affiliation: aDepartment of Neurology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu, China bUniversity of Rochester School of Medicine and Dentistry, Rochester, New York, USA.

ABSTRACT
(-)-Epigallocatechin-3-gallate (EGCG), the major polyphenolic component of green tea, has anti-inflammatory and antioxidant properties and provides neuroprotection against central nervous system diseases. Yet, it is not known whether EGCG may be neuroprotective against intracerebral hemorrhage. In this study, we used a simplified in-vitro model of thrombin neurotoxicity to test whether EGCG provides neuroprotection against thrombin-associated toxicity. Exposure of primary cortical neurons to thrombin (100 U/ml) caused dose-dependent and time-dependent cytotoxicity. Cell Counting Kit 8 and lactate dehydrogenase were used to monitor cell viability after exposure of neurons to thrombin or EGCG and after EGCG pretreatment. Flow cytometric analysis and western blotting demonstrated that thrombin-induced neuron degeneration occurs through apoptosis. A concentration of 25 μM EGCG significantly abolished thrombin-induced toxicity and prevented apoptosis by suppressing c-Jun-N-terminal kinase (JNK) phosphorylation, and the JNK inhibitor SP600125 reduced thrombin-induced caspase 3 activation and apoptosis. These data suggest that EGCG may have protective effects against thrombin-induced neuroapoptosis by inhibiting the activation of JNK, leading to caspase 3 cleavage. EGCG is a novel candidate neuroprotective agent against intracerebral hemorrhage-induced neurotoxicity.

Show MeSH

Related in: MedlinePlus

Assessment of neuron purity. (a) β3-Tublin-marked neurons showed red fluorescence. (b) Nuclei counterstained with DAPI showed blue fluorescence. (c) Colocalization of cytoplasm β3-tublin and nuclear DAPI (scale bar, 50 μm). DAPI, 4',6-diamidino-2-phenylindole.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4390119&req=5

Figure 1: Assessment of neuron purity. (a) β3-Tublin-marked neurons showed red fluorescence. (b) Nuclei counterstained with DAPI showed blue fluorescence. (c) Colocalization of cytoplasm β3-tublin and nuclear DAPI (scale bar, 50 μm). DAPI, 4',6-diamidino-2-phenylindole.

Mentions: Perikarya and dendrites of rat cortical neurons were labeled with a neuron-specific antibody, β3-tublin, and stained red. The nuclei were stained blue with DAPI. Neurons were seen to comprise roughly 90% of all nucleated cells (Fig. 1).


The protective role of (-)-epigallocatechin-3-gallate in thrombin-induced neuronal cell apoptosis and JNK-MAPK activation.

He Q, Bao L, Zimering J, Zan K, Zhang Z, Shi H, Zu J, Yang X, Hua F, Ye X, Cui G - Neuroreport (2015)

Assessment of neuron purity. (a) β3-Tublin-marked neurons showed red fluorescence. (b) Nuclei counterstained with DAPI showed blue fluorescence. (c) Colocalization of cytoplasm β3-tublin and nuclear DAPI (scale bar, 50 μm). DAPI, 4',6-diamidino-2-phenylindole.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390119&req=5

Figure 1: Assessment of neuron purity. (a) β3-Tublin-marked neurons showed red fluorescence. (b) Nuclei counterstained with DAPI showed blue fluorescence. (c) Colocalization of cytoplasm β3-tublin and nuclear DAPI (scale bar, 50 μm). DAPI, 4',6-diamidino-2-phenylindole.
Mentions: Perikarya and dendrites of rat cortical neurons were labeled with a neuron-specific antibody, β3-tublin, and stained red. The nuclei were stained blue with DAPI. Neurons were seen to comprise roughly 90% of all nucleated cells (Fig. 1).

Bottom Line: Flow cytometric analysis and western blotting demonstrated that thrombin-induced neuron degeneration occurs through apoptosis.These data suggest that EGCG may have protective effects against thrombin-induced neuroapoptosis by inhibiting the activation of JNK, leading to caspase 3 cleavage.EGCG is a novel candidate neuroprotective agent against intracerebral hemorrhage-induced neurotoxicity.

View Article: PubMed Central - PubMed

Affiliation: aDepartment of Neurology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu, China bUniversity of Rochester School of Medicine and Dentistry, Rochester, New York, USA.

ABSTRACT
(-)-Epigallocatechin-3-gallate (EGCG), the major polyphenolic component of green tea, has anti-inflammatory and antioxidant properties and provides neuroprotection against central nervous system diseases. Yet, it is not known whether EGCG may be neuroprotective against intracerebral hemorrhage. In this study, we used a simplified in-vitro model of thrombin neurotoxicity to test whether EGCG provides neuroprotection against thrombin-associated toxicity. Exposure of primary cortical neurons to thrombin (100 U/ml) caused dose-dependent and time-dependent cytotoxicity. Cell Counting Kit 8 and lactate dehydrogenase were used to monitor cell viability after exposure of neurons to thrombin or EGCG and after EGCG pretreatment. Flow cytometric analysis and western blotting demonstrated that thrombin-induced neuron degeneration occurs through apoptosis. A concentration of 25 μM EGCG significantly abolished thrombin-induced toxicity and prevented apoptosis by suppressing c-Jun-N-terminal kinase (JNK) phosphorylation, and the JNK inhibitor SP600125 reduced thrombin-induced caspase 3 activation and apoptosis. These data suggest that EGCG may have protective effects against thrombin-induced neuroapoptosis by inhibiting the activation of JNK, leading to caspase 3 cleavage. EGCG is a novel candidate neuroprotective agent against intracerebral hemorrhage-induced neurotoxicity.

Show MeSH
Related in: MedlinePlus