3-(Naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride attenuates MPP＋-induced cytotoxicity by regulating oxidative stress and mitochondrial dysfunction in SH-SY5Y cells

Abstract

Parkinson's disease (PD) is a common chronic neurodegenerative disease mainly caused by the death of dopaminergic neurons. However, no complete pharmacotherapeutic approaches are currently available for PD therapies. 1-methyl-4- phenylpyridinium (MPP＋)-induced SH-SY5Y neurotoxicity has been broadly utilized to create cellular models and study the mechanisms and critical aspects of PD. In the present study, we examined the role of a novel azetidine derivative, 3-(naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride (KHG26792), against MPP＋-induced neurotoxicity in SH-SY5Y cells. Treatment of KHG26792 significantly attenuated MPP＋-induced changes in the protein levels of Bcl-2 and Bax together with efficient suppression of MPP＋-induced activation of caspase-3 activity. KHG26792 also attenuated mitochondrial potential and levels of ROS, Ca2＋, and ATP in MPP＋-treated SH-SY5Y cells. Additionally, KHG26792 inhibited the induced production of nitric oxide and malondialdehyde. Moreover, the protective effect of KHG26792 is mediated through regulation of glutathione peroxidase and GDNF levels. Our results suggest a possibility that KHG26792 treatment significantly protects against MPP＋-induced neurotoxicity in SH-SY5Y cells and KHG26792 may be a valuable therapeutic agent for the treatment of PD induced by an environmental toxin. [BMB Reports 2018; 51(11): 590-595].

Fig. 1

Effects of KHG26792 on MPP⁺-induced cell death in SH-SY5Y cells. SH-SY5Y cells were pretreated with KHG26792 (0, 1, 5, 20, or 50 μM) for 1 h, followed by MPP⁺ (2 mM) for 24 h. (A) Chemical structure of KHG26792. (B) MTT assay. (C) LDH assay.

Fig. 2

Effects of KHG26792 on MPP⁺-induced imbalance in the expression of Bax and Bcl-2, and caspase-3 activity. (A) Total cell extracts were subjected to Western blotting analysis with antibodies against Bcl-2 and Bax. β-actin was used as a loading control. (B, C) Densitometric analysis is shown for the relative expression level of each protein. (D) Caspase-3 activity. Data from three independent experiments are presented as the means ± SD (*P < 0.05 relative to MPP⁺-treated cells).

Fig. 3

Effects of KHG26792 on mitochondrial potential, and levels of ROS, Ca²⁺, and ATP in MPP⁺-treated SH-SY5Y cells. SH-SY5Y cells were treated as described in Fig. 1. Mitochondrial membrane potential (A), ROS (B), Ca²⁺ (C), and ATP (D) levels were measured as described in Materials and Methods section. Data from three independent experiments are presented as the means ± SD (*P < 0.05 relative to MPP⁺-treated cells).

Fig. 4

Effects of KHG26792 on the levels of NO, MDA, GPx, and GDNF in MPP⁺-treated SH-SY5Y cells. SH-SY5Y cells were treated as described in Fig. 1. NO (A), MDA (B), GPx activity (C), and GDNF (D) levels were measured as described in Materials and Methods section. Data from three independent experiments are presented as the means ± SD (*P < 0.05 relative to MPP⁺-treated cells).