Rho kinase inhibitor Y-27632 down-regulates norepinephrine synthesis and release in PC12 cells.
Rho kinase inhibition is beneficial for neurite outgrowth and nerve disorders, and the Rho kinase inhibitors have been regarded as promising agents to treat neural diseases. The main aim of the study was to elucidate how Rho kinase inhibitor Y-27632 regulates neurotransmitter norepinephrine synthesis and release in PC12 cells when neurite outgrowth was induced. PC12 cells were treated with Y-27632 for 6 days. The amount of norepinephrine synthesized in PC12 cells and the amount released evoked by acetylcholine or by KCl were determined by norepinephrine enzyme-linked immunosorbent assay kits. The results showed that the amount of norepinephrine both synthesized and released was down-regulated with a concentration-dependent relationship. Further results of Western blotting found that the protein expression of tyrosine hydroxylase and synapsin I (especially its active form, synapsin I phosphoSer603) was also down-regulated, which were directly related to synthesis and release of norepinephrine, respectively. All the results suggest that Y-27632 is able to down-regulate norepinephrine synthesis and release, the direct mechanism of which may be associated with down-regulation on expression of some proteins, including tyrosine hydroxylase and synapsin I.
Duan
Wei-Gang
WG
Jiangsu Center for Drug Screening, Jiangsu Center for Pharmacodynamics Research and Evaluation, China Pharmaceutical University, Nanjing, China.
Shang
Jing
J
Jiang
Zhen-Zhou
ZZ
Yao
Jin-Cheng
JC
Yun
Yu
Y
Yan
Ming
M
Shu
Bin
B
Lin
Qing
Q
Yu
Ze-Pu
ZP
Zhang
Lu-Yong
LY
D000109:Acetylcholine / Q000494:pharmacology; D000577:Amides / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004797:Enzyme-Linked Immunosorbent Assay; D020932:Nerve Growth Factor / Q000494:pharmacology; D016501:Neurites / Q000187:drug effects* / Q000378:metabolism; D009638:Norepinephrine / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D016716:PC12 Cells; D011189:Potassium Chloride / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology*; D051381:Rats; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
D000577:Amides; D011725:Pyridines; C108830:Y 27632; D011189:Potassium Chloride; D020932:Nerve Growth Factor; D054460:rho-Associated Kinases; D000109:Acetylcholine; D009638:Norepinephrine