Comparative Proteome Analysis of Tumor Necrosis Factor
Alpha-Stimulated Human Vascular Smooth Muscle Cells in Response to Melittin
Proteome Sci, 2013 May 7;11(1):20
BACKGROUND:
Bee venom has been used to relieve pain and to treat
inflammatory diseases, including rheumatoid arthritis, in humans. To better
understand the mechanisms of the anti-inflammatory and anti-atherosclerosis
effect of bee venom, gel electrophoresis and mass spectrometry were used to
identify proteins whose expression was altered in human Vascular Smooth Muscle
Cells (hVSMCs) stimulated by tumor necrosis factor alpha after 12 h in the
presence of melittin.
RESULTS:
To obtain valuable insights into the anti-inflammatory and
anti-atherosclerosis mechanisms of melittin, two-dimensional (2-D) gel
electrophoresis and MALDI-TOF/TOF were used. The proteome study, we showed 33
significant proteins that were differentially expressed in the cells treated
with tumor necrosis factor alpha and melittin. Thirteen proteins were
significantly increased in the cells treated with tumor necrosis factor alpha,
and those proteins were reduced in the cells treated with melittin. Five of the
proteins that showed increased expression in the cells treated with tumor
necrosis factor alpha are involved in cell migration, including calreticulin,
an essential factor of development that plays a role in transcription
regulation. The proteins involved in cell migration were reduced in the melittin
treated cells. The observed changes in the expression of GRP75, prohibitin, and
a select group of other proteins were validated with reverse transcribed-PCR.
It was confirmed that the observed change in the protein levels reflected a
change in the genes level. In addition, the phosphorylation of EGFR and ERK was
validated by analyzing the protein pathway.
CONCLUSION:
Taken together, these data established that the expression
of some proteins was significantly changed by melittin treatment in tumor necrosis
factor alpha stimulated the cells and provided insights into the mechanism of
the melittin function for its potential use as an anti-inflammatory agent.
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