Protective effect of tubuloside B on TNFalpha-induced apoptosis in neuronal cells.

Abstract

AIM:

To investigate the neuroprotective effect of tubuloside B, one of the phenylethanoids isolated from the stems of Cistanche salsa, on tumor necrosis factor-alpha (TNFalpha)-induced apoptosis in SH-SY5Y neuronal cells.

METHODS:

Cell viability was analyzed using MTT assay. Apoptotic cells were detected using Hoechst33342 staining, and confirmed by DNA fragmentation and flow cytometric analysis. The activity of caspase-3 was measured with special assay kit. The concentration of free intracellular calcium was determined with the probe Indo-1 by spectrometer. The level of intracellular reactive oxygen species and the potential of mitochondrial membrane were determined by laser scanning confocal microscopy (LSCM) combined with fluorescence probe H2DCFDA or JC-1 respectively.

RESULTS:

SH-SY5Y cells treated with TNFalpha 100 microg/L for 36 h showed typical morphological changes of apoptosis. DNA ladder could be observed by agarose gel electrophoresis. The highest percentage of apoptotic cells accumulated to 37.5 %. Following 36 h treatment with TNFalpha, accumulation of intracellular ROS and [Ca2+]i and decrease in mitochondrial membrane potential were observed, and caspase-3 activity increased by about five-fold compared with controls. However, pretreatment with tubuloside B (1, 10, or 100 mg/L) for 2 h attenuated the TNFalpha-mediated apoptosis. The antiapoptotic action of tubuloside B was partially dependent on an anti-oxidative stress effects, maintain of mitochondria function, decrease of concentration of free intracellular calcium and inhibition of caspase-3 activity.

CONCLUSION:

Tubuloside B has the neuroprotective capacity to antagonize TNFalpha-induced apoptosis in SH-SY5Y cells and may be useful in treating some neurodegenerative diseases.

• Authors: Min Deng, Jin-Yuan Zhao, Xiao-Dong Ju, Peng-Fei Tu, Yong Jiang, Zheng-Bin Li
• Journal: Acta pharmacologica Sinica (Acta Pharmacol Sin) Vol. 25 Issue 10 Pg. 1276-84 (Oct 2004) ISSN: 1671-4083 [Print] United States
• PMID: 15456528 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright 2004 Acta Pharmacologica Sinica

Chemical References

• Glucosides
• Neuroprotective Agents
• Reactive Oxygen Species
• Tumor Necrosis Factor-alpha
• tubuloside B
• CASP3 protein, human
• Caspase 3
• Caspases
• Calcium

Topics

• Apoptosis (drug effects)
• Calcium (metabolism)
• Caspase 3
• Caspases (metabolism)
• Cell Line, Tumor
• Cistanche (chemistry)
• DNA Fragmentation (drug effects)
• Glucosides (isolation & purification, pharmacology)
• Humans
• Mitochondria (metabolism)
• Neuroblastoma (pathology)
• Neuroprotective Agents (pharmacology)
• Plants, Medicinal (chemistry)
• Reactive Oxygen Species (metabolism)
• Tumor Necrosis Factor-alpha (antagonists & inhibitors)