Objective: To establish a Parkinson's disease (PD) cell model of human neuroblastoma cells (SH-SY5Y) induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and to explore the possible mechanism of trehalose in PD. Methods: SH-SY5Y cells were cultured and divided into four groups: control group, trehalose group, MPTP group and trehalose +MPTP group.The trehalose was detected with MTT assay for its effect of on the viability of injured SH-SY5Y cells. The lactate dehydrogenase (LDH) was detected by kit for its level in culture medium. The ROS was assessed by DCFH-DA for its average level in cells. Besides, Hoechst 33342 staining was used to detect apoptosis. The endoplasmic reticulum stress (ERS) related proteins GRP78, IRE1, PERK and ATF6 were also detected by Western blot for its expression levels. Results: The survival rate of MPTP group decreased significantly and the levels of LDH and ROS increased compared with the control group. In addition, the survival rate of cells in trehalose +MPTP group increased significantly and the levels of LDH and ROS decreased compared with MPTP group. The expressions of GRP78, IRE1, p- IRE1, PERK, p- PERK and ATF6 in MPTP group increased, but decreased in trehalose +MPTP group compared with the control group. Conclusion: Trehalose has a significant protective effect on MPTP-induced SH-SY5Y cells injury, and its mechanism may be related to the effect of trehalose on oxidative stress by inhibiting ER stress response.
The goal of this study was to explore the regulatory mechanism of trehalose on the autophagy death of SH-SY5Y cells induced by oxygen and sugar deprivation from the aspect of glycolysis. Human SH-SY5Y cells were divided into three groups includes control group (control), model group (oxygen and sugar deprivation) and experimental group (oxygen and sugar deprivation+trehalose). The proliferation activity, mortality of SH-SY5Y cells, adenosine triphosphate and pyruvic acid were measured. Western blot was carried out to detect the expression levels of autophagic-related proteins (Unc-51 like autophagy activating kinase 1, beclin 1, autophagy related 5, microtubule-associated protein 1A/1B-light chain 3, ubiquitin-binding protein p62), glycolysis-related proteins (hexokinase 2, phosphofructokinase, pyruvate kinase M2) and activated protein kinase signalling pathway proteins. Compared with the control group, the autophagy level of the oxygen and sugar deprivation group was increased in a time-dependent manner (p<0.05). Cell mortality was increased (p<0.05) and the expressions of hexokinase 2, phosphofructokinase, pyruvate kinase M2 were decreased (p<0.05). The expression levels of activated protein kinase signalling pathway related proteins was upregulated (p<0.05). On the contrary, the cell death rate and autophagy level of the experimental group were significantly lower than that of the model group. The protein levels of hexokinase 2, phosphofructokinase, pyruvate kinase M2 were up-regulated and the difference was statistically significant (p<0.05). Trehalose can reduce the damage of SH-SY5Y cells caused by oxygen and sugar deprivation. The mechanism may be related to the improvement of glycolysis dysfunction and alleviation of autophagy over activation of activated protein kinase.
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