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Int J Mol Epidemiol Genet 2011;2(4):409-415
Original Article
The influence of tumour necrosis factor- α (TNF-α) on amyloid-β (Aβ)-degrading
enzymes in vitro
Doris Culpan, Jennifer Palmer, J Scott Miners, Seth Love, Patrick G Kehoe
Dementia Research Group, School of Clinical Sciences, University of Bristol, John James Building, Frenchay Hospital,
Bristol, BS16 1LE, United Kingdom.
Received November 21, 2011; accepted November 28, 2011; Epub November 30, 2011; Published December 15, 2011
Abstract: Pro-inflammatory cytokines, such as tumour necrosis factor-α (TNF-α), are increased in serum and CSF in
Alzheimer's disease (AD). We investigated the effect of TNF-α on gene and protein expression levels of Aβ degrading enzymes
(ACE, ECE-1, ECE- 2, IDE and NEP) in vitro. Differentiated (DC) and non-differentiated (NDC) neuroblastoma cells (SH-SY5Y)
were exposed to TNF-α for 15 minutes and 3 hours and protein and gene expression levels measured using western blotting
or sandwich ELISA (ECE-2), and real time-PCR (RT-PCR). Only ECE-2 protein levels decreased significantly in NDCs in a
dose-dependent manner after 15 minutes of TNF-α exposure but reverted to basal levels after 3 hours. Basal NEP gene
expression levels were higher in control DCs compared to NDCs but TNF-α treatment did not significantly alter the levels of
expression of any of the Aβ degrading enzymes. In conclusion, apart from a transient reduction in ECE-2 protein levels, TNF-α
had no impact in our in vitro experimental system on transcription or translation of any of our selected mediators of Aβ
degradation. (IJMEG1111005).
Key words: Alzheimer’s disease, inflammation, TNF-α, SH-SY5Y cells, RT-PCR, western blot
Full Text PDF
Address all correspondence to:
Dr. Doris Culpan, Dementia
Research Group, School of Clinical Sciences
University of Bristol, John James Building
Frenchay Hospital, Frenchay, Bristol, BS16 1LE, United Kingdom
Tel: 0117 3403068; Fax: 0117 957 3955;
Email: Doris.Culpan@bristol.ac.uk
Original Article
The influence of tumour necrosis factor- α (TNF-α) on amyloid-β (Aβ)-degrading
enzymes in vitro
Doris Culpan, Jennifer Palmer, J Scott Miners, Seth Love, Patrick G Kehoe
Dementia Research Group, School of Clinical Sciences, University of Bristol, John James Building, Frenchay Hospital,
Bristol, BS16 1LE, United Kingdom.
Received November 21, 2011; accepted November 28, 2011; Epub November 30, 2011; Published December 15, 2011
Abstract: Pro-inflammatory cytokines, such as tumour necrosis factor-α (TNF-α), are increased in serum and CSF in
Alzheimer's disease (AD). We investigated the effect of TNF-α on gene and protein expression levels of Aβ degrading enzymes
(ACE, ECE-1, ECE- 2, IDE and NEP) in vitro. Differentiated (DC) and non-differentiated (NDC) neuroblastoma cells (SH-SY5Y)
were exposed to TNF-α for 15 minutes and 3 hours and protein and gene expression levels measured using western blotting
or sandwich ELISA (ECE-2), and real time-PCR (RT-PCR). Only ECE-2 protein levels decreased significantly in NDCs in a
dose-dependent manner after 15 minutes of TNF-α exposure but reverted to basal levels after 3 hours. Basal NEP gene
expression levels were higher in control DCs compared to NDCs but TNF-α treatment did not significantly alter the levels of
expression of any of the Aβ degrading enzymes. In conclusion, apart from a transient reduction in ECE-2 protein levels, TNF-α
had no impact in our in vitro experimental system on transcription or translation of any of our selected mediators of Aβ
degradation. (IJMEG1111005).
Key words: Alzheimer’s disease, inflammation, TNF-α, SH-SY5Y cells, RT-PCR, western blot
Full Text PDF
Address all correspondence to:
Dr. Doris Culpan, Dementia
Research Group, School of Clinical Sciences
University of Bristol, John James Building
Frenchay Hospital, Frenchay, Bristol, BS16 1LE, United Kingdom
Tel: 0117 3403068; Fax: 0117 957 3955;
Email: Doris.Culpan@bristol.ac.uk
