A novel therapeutic approach to 6-OHDA-induced Parkinson's disease in rats via supplementation of PTD-conjugated tyrosine hydroxylase.
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.
Wu
Shao Ping
SP
Department of Biochemistry and Pharmacology, Beijing Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Tai Ping Road, 27, Beijing 100850, China.
Fu
Ai Ling
AL
Wang
Yu Xia
YX
Yu
Lei Ping
LP
Jia
Pei Yuan
PY
Li
Qian
Q
Jin
Guo Zhang
GZ
Sun
Man Ji
MJ
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
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