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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.

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Activation of JNK and caspase 3 in neuron cells. (a) The time dependence of JNK/caspase 3 activation in thrombin-induced neurons (*P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 vs. control group, ##P<0.05 vs. thrombin group). (b) EGCG pretreatment significantly decreased JNK and caspase 3 activation. (c) SP600125 pretreatment significantly decreased caspase 3 activation (*P<0.05 vs. thrombin group). Columns represent the mean±SE. n=3. DMSO, dimethyl sulfoxide; EGCG, (−)-epigallocatechin-3-gallate; JNK, c-Jun-N-terminal kinase; TM, thrombin.
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Figure 3: Activation of JNK and caspase 3 in neuron cells. (a) The time dependence of JNK/caspase 3 activation in thrombin-induced neurons (*P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 vs. control group, ##P<0.05 vs. thrombin group). (b) EGCG pretreatment significantly decreased JNK and caspase 3 activation. (c) SP600125 pretreatment significantly decreased caspase 3 activation (*P<0.05 vs. thrombin group). Columns represent the mean±SE. n=3. DMSO, dimethyl sulfoxide; EGCG, (−)-epigallocatechin-3-gallate; JNK, c-Jun-N-terminal kinase; TM, thrombin.

Mentions: Compared with the control group, the expression of phospho-JNK and cleaved-caspase 3 in the thrombin-treated group was increased. However, in the SP600125 or EGCG pretreated group, phospho-JNK and cleaved-caspase 3 levels were lower than in nonpretreated neurons (Fig. 3a–c).


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)

Activation of JNK and caspase 3 in neuron cells. (a) The time dependence of JNK/caspase 3 activation in thrombin-induced neurons (*P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 vs. control group, ##P<0.05 vs. thrombin group). (b) EGCG pretreatment significantly decreased JNK and caspase 3 activation. (c) SP600125 pretreatment significantly decreased caspase 3 activation (*P<0.05 vs. thrombin group). Columns represent the mean±SE. n=3. DMSO, dimethyl sulfoxide; EGCG, (−)-epigallocatechin-3-gallate; JNK, c-Jun-N-terminal kinase; TM, thrombin.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 3: Activation of JNK and caspase 3 in neuron cells. (a) The time dependence of JNK/caspase 3 activation in thrombin-induced neurons (*P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001 vs. control group, ##P<0.05 vs. thrombin group). (b) EGCG pretreatment significantly decreased JNK and caspase 3 activation. (c) SP600125 pretreatment significantly decreased caspase 3 activation (*P<0.05 vs. thrombin group). Columns represent the mean±SE. n=3. DMSO, dimethyl sulfoxide; EGCG, (−)-epigallocatechin-3-gallate; JNK, c-Jun-N-terminal kinase; TM, thrombin.
Mentions: Compared with the control group, the expression of phospho-JNK and cleaved-caspase 3 in the thrombin-treated group was increased. However, in the SP600125 or EGCG pretreated group, phospho-JNK and cleaved-caspase 3 levels were lower than in nonpretreated neurons (Fig. 3a–c).

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.

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Related in: MedlinePlus