title: Inhibition of ROCK2 expression protects against methamphetamine-induced neurotoxicity in PC12 cells.
abstract: Methamphetamine is a type of psychoactive drug. It is well known that neurotoxicity caused by Methamphetamine(METH) can damage the nervous system and lead to apoptosis and cell loss of dopaminergic neurons. ROCK2 is a prominent target for gene therapy because its inhibition has proved to have a protective effect in various cell lines and pathophysiological conditions. Although several of the negative effects of METH on the dopaminergic system have been studied, the protective molecular mechanisms and the effective treatment of METH-induced apoptosis remain to be clarified. We hypothesized that ROCK2 is involved in METH-induced apoptosis. We tested our hypothesis using RT-PCR and western blotting to analyze whether silencing of ROCK2 with small interfering RNA (siROCK2) could reduce damage and apoptosis in PC12 cells after METH exposure. Increases in viability and cytomorphological changes were detected by MTT assay and bright field microscopy after pretreatment of METH-treated PC12 cells with 100 nM siROCK2. Apoptosis decreased significantly after ROCK2 silencing, as shown by Annexin V and TUNEL staining. The results show that ROCK2 is a possible gene target for therapeutics in METH-induced neurotoxicity in vitro, providing a foundation for future in vivo research.
authors: Institute of Forensic Medicine, Southern Medical University, Guangzhou 510515, China.
authors: Xingyi
authors: X
authors: Yang
authors: Yunyun
authors: Y
authors: Liu
authors: Chao
authors: C
authors: Liu
authors: Weibing
authors: W
authors: Xie
authors: Enping
authors: E
authors: Huang
authors: Weiye
authors: W
authors: Huang
authors: Jiawen
authors: J
authors: Wang
authors: Ling
authors: L
authors: Chen
authors: Huipin
authors: H
authors: Wang
authors: Pingming
authors: P
authors: Qiu
authors: Jingtao
authors: J
authors: Xu
authors: Fu
authors: F
authors: Zhang
authors: Huijun
authors: H
authors: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D008694:Methamphetamine / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology*; D016716:PC12 Cells; D051381:Rats; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D008694:Methamphetamine; C580818:ROCK2 protein, rat; D054460:rho-Associated Kinases
keywords: 5-HT; 5-hydroxytryptamine; Apoptosis; DA; Dopamine; FITC; Lipo; Lipofectamine 2000; METH; Methamphetamine; Neurotoxicity; PD; Parkinson's disease; ROCK2; Rho-associated kinase II; Rho-associated kinases II; Small interfering RNA; TH; Tyrosine hydroxylase; fluorescein isothiocyanate; methamphetamine; siNC; siRNA; siROCK2; small interfering RNA; small interfering RNA of ROCK2; small interfering nonspecific control