title: Expression profiles of hippocampal regenerative sprouting-related genes and their regulation by E-64d in a developmental rat model of penicillin-induced recurrent epilepticus.
abstract: E-64d (a calpain and autophagy inhibitor) has previously been shown safe for the treatment of Alzheimer's disease in humans. In the present study, the potential protective mechanism of E-64d on hippocampal aberrant mossy fiber sprouting was examined in a developmental rat model of penicillin-induced recurrent epilepticus. A seizure was induced by penicillin every other day in Sprague-Dawley rats from postnatal day 21 (P21). The rats were randomly assigned into the control group (CONT1), the control plus E-64d (CONT2), the seizure group (EXP1) and the seizure plus E-64d (EXP2). On P51, mossy fiber sprouting and related gene expression in hippocampus were assessed by Timm staining and real-time RT-PCR methods, respectively. To validate the RT-PCR results, western blot analysis was performed on selected genes. E-64d obviously suppressed the aberrant mossy fiber sprouting in the supragranular region of dentate gyrus and CA3 subfield of hippocampus. Among the total twelve genes, six genes were strongly up- (MT-3, ACAT1, clusterin and ApoE) or down- (ZnT-1 and PRG-3) regulated by developmental seizures (EXP1) compared with that in the CONT1. Up-regulation of ApoE and Clusterin was blocked by pretreatment with E-64d both in mRNA and protein levels. Further, E-64d-pretreated seizure rats (EXP2) showed a significant downregulation of mRNA expression of PRG-1, PRG-3 and PRG-5, cathepsin B and ApoE, as well as up-regulated nSMase and ANX7 in hippocampus when compared with EXP1 rats. The results of the present study suggest that E-64d, an elective inhibitor of calpain and autophagy, is potentially useful in the treatment of developmental seizure-induced brain damage both by regulating abnormal zinc signal transduction and through the modulation of altered lipid metabolism via ApoE/clusterin pathway in hippocampus.
authors.affiliation: Neurology Laboratory, Soochow University Affiliated Children's Hospital, Laboratory of Aging and Nervous Diseases, Soochow University, Suzhou 215003, China. nhdoctor@163.com
authors.forename: Hong
authors.initials: H
authors.lastname: Ni
authors.forename: Shou-Yun
authors.initials: SY
authors.lastname: Ren
authors.forename: Le-Ling
authors.initials: LL
authors.lastname: Zhang
authors.forename: Qi
authors.initials: Q
authors.lastname: Sun
authors.forename: Tian
authors.initials: T
authors.lastname: Tian
authors.forename: Xing
authors.initials: X
authors.lastname: Feng
mesh_terms: D000101:Acetyl-CoA C-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D017302:Annexins / Q000235:genetics / Q000378:metabolism; D002403:Cathepsins / Q000235:genetics / Q000378:metabolism; D027682:Cation Transport Proteins / Q000235:genetics / Q000378:metabolism; D051152:Clusterin / Q000235:genetics / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D004195:Disease Models, Animal; D005786:Gene Expression Regulation / Q000187:drug effects*; D007930:Leucine / Q000031:analogs & derivatives* / Q000494:pharmacology; D008297:Male; D000090023:Metallothionein 3; D019599:Mossy Fibers, Hippocampal / Q000187:drug effects* / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D012640:Seizures / Q000139:chemically induced / Q000235:genetics* / Q000378:metabolism
chemical_list: C568306:(3-ethoxycarbonyloxirane-2-carbonyl)leucine (3-methylbutyl) amide; D017302:Annexins; D027682:Cation Transport Proteins; C498500:Clu protein, rat; D051152:Clusterin; D015853:Cysteine Proteinase Inhibitors; D000090023:Metallothionein 3; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C092392:Slc30a1 protein, rat; D000101:Acetyl-CoA C-Acetyltransferase; D002403:Cathepsins; D007930:Leucine