Japanese encephalitis virus (JEV), which causes neurological disorders, completes its life cycle and triggers apoptotic cell death in infected cells. Dehydroepiandrosterone (DHEA), an adrenal-derived steroid, has been implicated in protection against neurotoxicity and protection of animals from viral-induced encephalitis, resulting in an increased survival rate of the animals. Currently, the mechanisms underlying the beneficial effects of DHEA against the virus are largely unknown. In this study, DHEA suppression of JEV replication and virus-induced apoptosis in murine neuroblastoma (N18) cells was investigated. It was found that DHEA suppressed JEV-induced cytopathic effects, JEV-induced apoptotic cell death and JEV propagation in a concentration-dependent manner. Antiviral activity was more efficient in cultures treated with DHEA immediately after viral adsorption compared with that in cultures receiving delayed administration after adsorption or transient exposure before adsorption. JEV-induced cytotoxicity was accompanied by the inactivation of extracellular signal-regulated protein kinase (ERK). Inactivation of ERK by JEV infection was reversed by DHEA. When cells were treated with the ERK inhibitor U0126, DHEA lost its antiviral effect. Activation of ERK by anisomycin mimicked the action of DHEA in suppressing JEV-induced cytotoxicity. DHEA-related compounds, such as its sulfate ester (DHEAS) and pregnenolone, were unable to suppress JEV-induced cytotoxicity and ERK inactivation. The hormone-receptor antagonists ICI 182780 and flutamide failed to abrogate the antiviral effect of DHEA. These findings suggest that the antiviral effect of DHEA is not linked directly to the genomic steroid-receptor pathways and suggest that the signalling pathways of ERK play a role in the antiviral action of DHEA.
BarberS. A.,
BruettL.,
DouglassB. R.,
HerbstD. S.,
ZinkM. C.,
ClementsJ. E.2002; Visna virus-induced activation of MAPK is required for virus replication and correlates with virus-induced neuropathology. J Virol 76:817–828[CrossRef]
BargerS. W.,
ChavisJ. A.,
DrewP. D.2000; Dehydroepiandrosterone inhibits microglial nitric oxide production in a stimulus-specific manner. J Neurosci Res 62:503–509[CrossRef]
Ben-NathanD.,
LustigS.,
KobilerD.,
DanenbergH. D.,
LupuE.,
FeuersteinG.1992; Dehydroepiandrosterone protects mice inoculated with West Nile virus and exposed to cold stress. J Med Virol 38:159–166[CrossRef]
BradleyW. G.,
KrausL. A.,
GoodR. A.,
DayN. K.1995; Dehydroepiandrosterone inhibits replication of feline immunodeficiency virus in chronically infected cells. Vet Immunol Immunopathol 46:159–168[CrossRef]
ChaturvediU. C.,
MathurA.,
TandonP.,
NatuS. M.,
RajvanshiS.,
TandonH. O.1979; Variable effect on peripheral blood leucocytes during JE virus infection of man. Clin Exp Immunol 38:492–498
ChenC.-J.,
RaungS.-L.,
KuoM.-D.,
WangY.-M.2002; Suppression of Japanese encephalitis virus infection by non-steroidal anti-inflammatory drugs. J Gen Virol 83:1897–1905
ChenC.-J.,
ChenJ.-H.,
ChenS.-Y.,
LiaoS.-L.,
RaungS.-L.2004; Upregulation of RANTES gene expression in neuroglia by Japanese encephalitis virus infection. J Virol 78:12107–12119[CrossRef]
CompagnoneN. A.,
MellonS. H.1998; Dehydroepiandrosterone: a potential signalling molecule for neocortical organization during development. Proc Natl Acad Sci U S A 95:4678–4683[CrossRef]
DaigleJ.,
CarrD. J. J.1998; Androstenediol antagonizes herpes simplex virus type 1-induced encephalitis through the augmentation of type I IFN production. J Immunol 160:3060–3066
DesaiA.,
ShankarS. K.,
RaviV.,
ChandramukiA.,
Gourie-DeviM.1995; Japanese encephalitis virus antigen in the human brain and its topographic distribution. Acta Neuropathol 89:368–373[CrossRef]
GordonG. B.,
BushD. E.,
WeismanH. F.1988; Reduction of atherosclerosis by administration of dehydroepiandrosterone. A study in the hypercholesterolemic New Zealand White rabbit with aortic intimal injury. J Clin Invest 82:712–720[CrossRef]
GordonG.,
MackowM. C.,
LevyH. R.1995; On the mechanism of interaction of steroids with human glucose 6-phosphate dehydrogenase. Arch Biochem Biophys 318:25–29[CrossRef]
HendersonE.,
YangJ. Y.,
SchwartzA.1992; Dehydroepiandrosterone (DHEA) and synthetic DHEA analogs are modest inhibitors of HIV-1 IIIB replication. AIDS Res Hum Retroviruses 8:625–631[CrossRef]
JacobsonM. A.,
FusaroR. E.,
GalmariniM.,
LangW.1991; Decreased serum dehydroepiandrosterone is associated with an increased progression of human immunodeficiency virus infection in men with CD4 cell counts of 200–499. J Infect Dis 164:864–868[CrossRef]
KonishiY.,
SatoH.,
TanakaT.2004; Anisomycin superinduces annexin V mRNA expression through the ERK1/2 but not the p38 MAP kinase pathway. Biochem Biophys Res Commun 313:977–983[CrossRef]
LiaoC.-L.,
LinY.-L.,
WangJ.-J.,
HuangY.-L.,
YehC.-T.,
MaS.-H.,
ChenL.-K.1997; Effect of enforced expression of human bcl-2 on Japanese encephalitis virus-induced apoptosis in cultured cells. J Virol 71:5963–5971
LiaoS.-L.,
RaungS.-L.,
ChenC.-J.2002; Japanese encephalitis virus stimulates superoxide dismutase activity in rat glial cultures. Neurosci Lett 324:133–136[CrossRef]
LoriaR. M.,
IngeT. H.,
CookS. S.,
SzakalA. K.,
RegelsonW.1988; Protection against acute lethal viral infections with the native steroid dehydroepiandrosterone (DHEA). J Med Virol 26:301–314[CrossRef]
LucasJ. A.,
AhmedS. A.,
CaseyM. L.,
MacDonaldP. C.1985; Prevention of autoantibody formation and prolonged survival in New Zealand Black/New Zealand White F1 mice fed dehydroisoandrosterone. J Clin Invest 75:2091–2093[CrossRef]
LuoH.,
YanagawaB.,
ZhangJ.7 other authors2002; Coxsackievirus B3 replication is reduced by inhibition of the extracellular signal-regulated kinase (ERK) signaling pathway. J Virol 76:3365–3373[CrossRef]
MajewskaM. D.1995; Neuronal action of dehydroepiandrosterone: possible roles in brain development, aging, memory, and affect. Ann N Y Acad Sci 774:111–120[CrossRef]
MajewskaM. D.,
DemirgorenD.,
SpivakC. E.,
LondonE. D.1990; The neurosteroid dehydroepiandrosterone sulfate is an allosteric antagonist of the GABAA receptor. Brain Res 526:143–146[CrossRef]
MathurA.,
BharadwajM.,
KulshreshthaR.,
RawatS.,
JainA.,
ChaturvediU. C.1988; Immunopathological study of spleen during Japanese encephalitis virus infection in mice. Br J Exp Pathol 69:423–432
MoralesA. J.,
NolanJ. J.,
NelsonJ. C.,
YenS. S. C.1994; Effects of replacement dose of dehydroepiandrosterone in men and women of advancing age. J Clin Endocrinol Metab 78:1360–1367
MulderJ. W.,
FrissenP. H.,
KrijnenP.,
EndertE.,
de WolfF.,
GoudsmitJ.,
MastersonJ. G.,
LangeJ. M.1992; Dehydroepiandrosterone as predictor for progression to AIDS in asymptomatic human immunodeficiency virus-infected men. J Infect Dis 165:413–418[CrossRef]
NestlerJ. E.,
BarlasciniC. O.,
CloreJ. N.,
BlackardW. G.1988; Dehydroepiandrosterone reduces serum low density lipoprotein levels and body fat but does not alter insulin sensitivity in normal men. J Clin Endocrinol Metab 66:57–61[CrossRef]
PleschkaS.,
WolffT.,
EhrhardtC.,
HobomG.,
PlanzO.,
RappU. R.,
LudwigS.2001; Influenza virus propagation is impaired by inhibition of the Raf/MEK/ERK signalling cascade. Nat Cell Biol 3:301–305[CrossRef]
RaoM. S.,
SubbaraoV.,
YeldandiA. V.,
ReddyJ. K.1992; Inhibition of spontaneous testicular Leydig cell tumor development in F-344 rats by dehydroepiandrosterone. Cancer Lett 65:123–126[CrossRef]
RaungS.-L.,
KuoM.-D.,
WangY.-M.,
ChenC.-J.2001; Role of reactive oxygen intermediates in Japanese encephalitis virus infection in murine neuroblastoma cells. Neurosci Lett 315:9–12[CrossRef]
RaviV.,
ParidaS.,
DesaiA.,
ChandramukiA.,
Gourie-DeviM.,
GrauG. E.1997; Correlation of tumor necrosis factor levels in the serum and cerebrospinal fluid with clinical outcome in Japanese encephalitis patients. J Med Virol 51:132–136[CrossRef]
ShihS.-R.,
TsaiK.-N.,
LiY.-S.,
ChuehC.-C.,
ChanE.-C.2003; Inhibition of enterovirus 71-induced apoptosis by allophycocyanin isolated from a blue-green alga Spirulina platensis
. J Med Virol 70:119–125[CrossRef]
SinghA.,
KulshreshthaR.,
MathurA.2000; Secretion of the chemokine interleukin-8 during Japanese encephalitis virus infection. J Med Microbiol 49:607–612
SolomonT.,
DungN. M.,
WillisB.9 other authors2003; Interferon alfa-2a in Japanese encephalitis: a randomised double-blind placebo-controlled trial. Lancet 361:821–826[CrossRef]
StanciuM.,
WangY.,
KentorR.8 other authors2000; Persistent activation of ERK contributes to glutamate-induced oxidative toxicity in a neuronal cell line and primary cortical neuron cultures. J Biol Chem 275:12200–12206[CrossRef]
UozumiK.,
UematsuT.,
OtsukaM.,
NakanoS.,
TakatsukaY.,
IwahashiM.,
HanadaS.,
ArimaT.1996; Serum dehydroepiandrosterone and DHEA-sulfate in patients with adult T-cell leukemia and human T-lymphotropic virus type I carriers. Am J Hematol 53:165–168[CrossRef]
van VollenhovenR. F.,
EnglemanE. G.,
McGuireJ. L.1994; An open study of dehydroepiandrosterone in systemic lupus erythematosus. Arthritis Rheum 37:1305–1310[CrossRef]
WangX.,
MartindaleJ. L.,
LiuY.,
HolbrookN. J.1998; The cellular response to oxidative stress: influences of mitogen-activated protein kinase signalling pathways on cell survival. Biochem J 333:291–300
WolfO. T.,
KirschbaumC.1999; Actions of dehydroepiandrosterone and its sulfate in the central nervous system: effects on cognition and emotion in animals and humans. Brain Res Brain Res Rev 30:264–288[CrossRef]
WuS.-F.,
LeeC.-J.,
LiaoC.-L.,
DwekR. A.,
ZitzmannN.,
LinY.-L.2002; Antiviral effects of an iminosugar derivative on flavivirus infections. J Virol 76:3596–3604[CrossRef]
YamashitaY.,
ItoT.,
HashimotoI.,
OhyamaA.,
KuriyamaK.1989; Changes in cerebral level of monoamines by Japanese encephalitis virus infection. Uirusu 39:47–54 (in Japanese [CrossRef]
YangN.-C.,
JengK.-C. G.,
HoW.-M.,
ChouS.-J.,
HuM.-L.2000; DHEA inhibits cell growth and induces apoptosis in BV-2 cells and the effects are inversely associated with glucose concentration in the medium. J Steroid Biochem Mol Biol 75:159–166[CrossRef]