title: A novel therapeutic approach to 6-OHDA-induced Parkinson's disease in rats via supplementation of PTD-conjugated tyrosine hydroxylase.
abstract: The present study aimed to evaluate whether the protein transduction domain (PTD)-conjugated human tyrosine hydroxylase (TH) fusion protein was effective on the 6-hydroxydopamine (6-OHDA)-induced Parkinson's disease (PD) model rats. An expression vector pET-PTD-TH harbouring the PTD-TH gene was constructed and transformed to the Escherichia coli BL21 cells for expression. The expressed recombinant PTD-TH with a molecular weight of 61 kD was successfully transduced (1 microM) into the dopaminergic SH-sy5y human neuroblastoma cells in vitro and visualized by immunohistochemical assay. An in vivo experiment in rats showed that the iv administered PTD-TH protein (8 mg/kg) permeated across the blood-brain barrier, penetrated into the striatum and midbrain, and peaked at 5-8 h after the injection. The behavioral effects of PTD-TH on the apomorphine-induced rotations in the PD model rats 8 weeks after the 6-OHDA lesion showed that a single bolus of PTD-TH (8 mg/kg) iv injection caused a decrement of 60% of the contralateral turns on day 1 and 40% on days 5-17. The results imply that iv delivery of PTD-TH is therapeutically effective on the 6-OHDA-induced PD in rats, the PTD-mediated human TH treatment opening a promising therapeutic direction in treatment of PD.
authors.affiliation: Department of Biochemistry and Pharmacology, Beijing Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Tai Ping Road, 27, Beijing 100850, China.
authors.forename: Shao Ping
authors.initials: SP
authors.lastname: Wu
authors.forename: Ai Ling
authors.initials: AL
authors.lastname: Fu
authors.forename: Yu Xia
authors.initials: YX
authors.lastname: Wang
authors.forename: Lei Ping
authors.initials: LP
authors.lastname: Yu
authors.forename: Pei Yuan
authors.initials: PY
authors.lastname: Jia
authors.forename: Qian
authors.initials: Q
authors.lastname: Li
authors.forename: Guo Zhang
authors.initials: GZ
authors.lastname: Jin
authors.forename: Man Ji
authors.initials: MJ
authors.lastname: Sun
mesh_terms: D000818:Animals; D001058:Apomorphine / Q000494:pharmacology; D001812:Blood-Brain Barrier; D001923:Brain Chemistry; D045744:Cell Line, Tumor; D015696:Gene Products, tat / Q000235:genetics; D006801:Humans; D008297:Male; D016627:Oxidopamine; D010302:Parkinson Disease, Secondary / Q000139:chemically induced / Q000188:drug therapy*; D010446:Peptide Fragments / Q000235:genetics; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000627:therapeutic use*; D012399:Rotation; D013239:Stereotyped Behavior / Q000187:drug effects; D014161:Transduction, Genetic; D014446:Tyrosine 3-Monooxygenase / Q000008:administration & dosage / Q000627:therapeutic use*; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D015696:Gene Products, tat; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus; C497137:tat peptide (47-57), Human immunodeficiency virus 1; D016627:Oxidopamine; D014446:Tyrosine 3-Monooxygenase; D001058:Apomorphine