LncRNA RPPH1 attenuates Aβ_25-35-induced endoplasmic reticulum stress and apoptosis in SH-SY5Y cells via miR-326/PKM2

Abstract

Background

The durative endoplasmic reticulum stress (ERS) and subsequent apoptosis contributes to the development and progression of Alzheimer’s disease (AD). MiR-326 can reduce pyruvate kinase M2 (PKM2) expression, leading to ERS. Whereas, lncRNA RPPH1 is able to increase dendritic spine density and protect hippocampal pyramidal neurons through targeting miR-326. Our study aims to investigate the regulation of lncRNA RPPH1 and miR-326/PKM2 on ERS and related apoptosis in AD.

Methods

SH-SY5Y cells treated with Aβ_25-35 were selected as an in vitro AD model. RPPH1 and miR-326 overexpression and silencing cells were established by transforming vectors. The expression of RPPH1 and miR-326 were detected by qRT-PCR. MTT, flow cytometric, intracellular calcium assay and Western blot were used to test the functions of RPPH1 and miR-326 in SH-SY5Y cell proliferation, apoptosis and ERS. Dual-luciferase assay was used to detect the interaction among RPPH1, miR-326 and PKM2.

Results

RPPH1 overexpression enhanced the viability of SH-SY5Y cells, and attenuated the apoptosis of of SH-SY5Y cells. Moreover, RPPH1 overexpression down-regulated ER stress related proteins such as GRP78, CHOP and cleaved caspase-12. Mechanistically, RPPH1 directly targeted miR-326, thereby counteracting its inhibitory effect on PKM2 expression, contributing to attenuation of apoptosis and ERS induced by Aβ_25-35.

Conclusion

Aβ_25-35-induced ERS and apoptosis in SH-SY5Y cells can be attenuated by lncRNA RPPH1 through regulating miR-326/PKM2 axis. This study provided therapeutic options for AD patients.

Keywords:

• Alzheimer’s disease
• lncRNA RPPH1
• miR-326
• endoplasmic reticulum stress
• apoptosis
• pyruvate kinase M2

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by Youth Science Foundation (Num: 81401083): Correlation between oligodendrocyte differentiation dysfunction and the pathogenesis of epilepsy and Joint Fund Project of Guizhou Province (Num: Qian Ke He LH [2014]7022): EPO participates in the neuroprotection of AngII through PI3-K/AKT and ERK1/2 signaling pathways.