title: Oxidative stress in NPC1 deficient cells: protective effect of allopregnanolone.
abstract: Niemann-Pick C disease (NPC) is an autosomal recessive neurodegenerative disorder caused by the abnormal function of NPC1 or NPC2 proteins, leading to an accumulation of unesterified cholesterol and glycosphingolipids (GSLs) in the lysosomes. The mechanisms underlying the pathophysiology in NPC disease are not clear. Oxidative damage is implicated in the pathophysiology of different neurological disorders and the effect of GSL accumulation on the intracellular redox state has been documented. Therefore, we determined whether the intracellular redox state might contribute to the NPC disease pathophysiology. Because the treatment of NPC mice with allopregnanolone (ALLO) increases their lifespan and delays the onset of neurological impairment, we analysed the effect of ALLO on the oxidative damage in human NPC fibroblasts. Concentrations of reactive oxygen species (ROS) and lipid peroxidation were higher in fibroblasts from NPC patients than in fibroblasts from normal subjects. Fibroblasts from NPC patients were more susceptible to cell death through apoptosis after an acute oxidative insult. This process is mediated by activation of the NF-kappaB signalling pathway. Knockdown of NPC1 mRNA both in normal fibroblasts and in human SH-SY5Y neuroblastoma cells caused increased ROS concentrations. ALLO treatment of fibroblasts from NPC patients or NPC1 knockdown cells reduced the levels of ROS and lipid peroxidation and prevented peroxide-induced apoptosis and NF-kB activation. Thus, these findings suggest that oxidative stress might contribute to the NPC disease and ALLO might be beneficial in the treatment of the disease, at least in part, due to its ability to restore the intracellular redox state.
authors.affiliation: Unita di Malattie Metaboliche, IRCCS Burlo Garofolo, Trieste, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Zampieri
authors.forename: Synthia H
authors.initials: SH
authors.lastname: Mellon
authors.forename: Terry D
authors.initials: TD
authors.lastname: Butters
authors.forename: Marco
authors.initials: M
authors.lastname: Nevyjel
authors.forename: Douglas F
authors.initials: DF
authors.lastname: Covey
authors.forename: Bruno
authors.initials: B
authors.lastname: Bembi
authors.forename: Andrea
authors.initials: A
authors.lastname: Dardis
mesh_terms: D017209:Apoptosis; D001706:Biopsy; D002352:Carrier Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D002675:Child, Preschool; D002784:Cholesterol / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D015227:Lipid Peroxidation; D008247:Lysosomes / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D000077296:Niemann-Pick C1 Protein; D052556:Niemann-Pick Disease, Type C / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011280:Pregnanolone / Q000494:pharmacology*
chemical_list: D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D002784:Cholesterol; D011280:Pregnanolone
references.citation: Clin Genet. 2003 Oct;64(4):269-81
references.pmid: 12974729
references.citation: Curr Alzheimer Res. 2006 Feb;3(1):11-7
references.pmid: 16472197
references.citation: Psychopharmacology (Berl). 2006 Jun;186(3):481-5
references.pmid: 16231167
references.citation: J Neurochem. 1999 Dec;73(6):2501-9
references.pmid: 10582611
references.citation: Trends Endocrinol Metab. 2001 May-Jun;12(4):152-6
references.pmid: 11295570
references.citation: Am J Hum Genet. 1996 Jan;58(1):118-25
references.pmid: 8554047
references.citation: Cell. 2001 Mar 9;104(5):719-30
references.pmid: 11257226
references.citation: PLoS One. 2006 Dec 20;1:e19
references.pmid: 17183645
references.citation: Science. 1997 Jul 11;277(5323):228-31
references.pmid: 9211849
references.citation: Science. 1986 May 23;232(4753):1004-7
references.pmid: 2422758
references.citation: Biochim Biophys Acta. 1980 Jan 18;617(1):97-109
references.pmid: 7353026
references.citation: J Neurochem. 2006 Aug;98(4):1278-89
references.pmid: 16771828
references.citation: J Cell Physiol. 2007 Jul;212(1):223-35
references.pmid: 17443679
references.citation: Blood. 1994 Sep 1;84(5):1415-20
references.pmid: 8068938
references.citation: J Med Genet. 1994 Apr;31(4):317-20
references.pmid: 8071958
references.citation: Trends Endocrinol Metab. 2002 Jan-Feb;13(1):35-43
references.pmid: 11750861
references.citation: Nat Med. 2004 Jul;10(7):704-11
references.pmid: 15208706
references.citation: Anal Biochem. 2004 Aug 15;331(2):275-82
references.pmid: 15265733
references.citation: Nature. 2001 Dec 13;414(6865):813-20
references.pmid: 11742414
references.citation: J Neurosci Res. 2005 Dec 15;82(6):811-21
references.pmid: 16273542
references.citation: Curr Alzheimer Res. 2006 Jul;3(3):185-90
references.pmid: 16842093
references.citation: J Biol Chem. 1997 Mar 7;272(10):6245-51
references.pmid: 9045641
references.citation: Biol Psychiatry. 2006 Dec 15;60(12):1287-94
references.pmid: 16997284
references.citation: Mol Cell Endocrinol. 2006 Feb 26;246(1-2):76-82
references.pmid: 16420972
references.citation: Biochim Biophys Acta. 2007 Jan;1768(1):131-45
references.pmid: 16945324
references.citation: Cancer Lett. 2005 Jan 31;218(1):1-14
references.pmid: 15639335
references.citation: Proc Natl Acad Sci U S A. 1988 Nov;85(21):8022-6
references.pmid: 3186703
references.citation: Free Radic Res Commun. 1991;12-13 Pt 2:783-94
references.pmid: 2060851
references.citation: Neurobiol Aging. 2005 Mar;26(3):293-302
references.pmid: 15639306
references.citation: Proc Natl Acad Sci U S A. 2003 Sep 30;100(20):11741-6
references.pmid: 14504383
references.citation: J Nutr Biochem. 2000 Nov;11(11-12):548-554
references.pmid: 11137891
references.citation: Steroids. 2002 May;67(6):471-5
references.pmid: 11960623
references.citation: Curr Alzheimer Res. 2006 Sep;3(4):269-83
references.pmid: 17017859
references.citation: J Clin Invest. 1993 Jun;91(6):2546-51
references.pmid: 8390482
references.citation: Antioxid Redox Signal. 1999 Summer;1(2):129-44
references.pmid: 11228742
references.citation: Proc Natl Acad Sci U S A. 2006 Sep 12;103(37):13807-12
references.pmid: 16940355
references.citation: Neurochem Res. 1999 Jan;24(1):69-77
references.pmid: 9973239
references.citation: Endocr Rev. 2007 Dec;28(7):726-41
references.pmid: 17916740
references.citation: J Steroid Biochem Mol Biol. 2002 Dec;83(1-5):195-7
references.pmid: 12650716
references.citation: Free Radic Biol Med. 2002 Nov 1;33(9):1231-42
references.pmid: 12398931
references.citation: Antioxid Redox Signal. 2006 Nov-Dec;8(11-12):2021-37
references.pmid: 17034347
references.citation: Mol Pharmacol. 1996 Dec;50(6):1581-6
references.pmid: 8967980