Silencing of LncRNA BDNF-AS attenuates Aβ25-35-induced neurotoxicity in PC12 cells by suppressing cell apoptosis and oxidative stress

Cong-Cong Guo; Chun-Hong Jiao; Zhen-Mei Gao

doi:10.1080/01616412.2018.1480921

Silencing of LncRNA BDNF-AS attenuates Aβ_25-35-induced neurotoxicity in PC12 cells by suppressing cell apoptosis and oxidative stress

Neurol Res. 2018 Sep;40(9):795-804. doi: 10.1080/01616412.2018.1480921. Epub 2018 Jun 14.

Authors

Cong-Cong Guo¹, Chun-Hong Jiao¹, Zhen-Mei Gao²

Affiliations

¹ a Department of rehabilitation , The People's Hospital of Zhangqiu , Ji'nan , China.
² b Department of rehabilitation , Affiliated Hospital of Shandong University of Traditional Chinese Medicine , Jinan , China.

PMID: 29902125
DOI: 10.1080/01616412.2018.1480921

Abstract

Objective: To explore the effects of long non-coding RNA (lncRNA) brain-derived neurotrophic factor anti-sense (BDNF-AS) on the Aβ_25-35-induced neurotoxicity in PC12 cells.

Methods: PC12 cells were induced by Aβ_25-35 to construct cell injury models of Alzheimer's disease (AD), and then transfected with siRNA-BDNF-AS. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to detect the expressions of BDNF-AS and BDNF. Besides, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Hoechst33342 staining were utilized to analyze the cell viability and apoptosis, respectively, Western blotting to evaluate the protein expressions, immunofluorescence to assess the Cytochrome C (Cyt C) release, and Rhodamine 123 (Rh123) to measure the mitochondrial membrane potential (MMP).The evaluation of oxidative stress was conducted via the determination of the levels of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT).

Results: Aβ_25-35 apparently increased BDNF-AS but decreased BDNF in PC12 cells, which also reduced viability and induced apoptosis of PC12 cells. Silencing of BDNF-AS could significantly up-regulate BDNF in Aβ_25-35-induced PC12 cells, with the elevated cell viability. Moreover, silencing BDNF-AS inhibited the apoptosis of Aβ_25-35-induced PC12 cells, suppressed the release of Cyt C, reduced the expression of cleaved caspase-3 and Bax, and lowered the mean fluorescence intensity (MFI) of Rh123, but it elevated the expression of Bcl-2. Besides, silencing BDNF-AS also reduced ROS intensity and MDA content, but enhanced the activities of SOD and CAT.

Conclusion: Silencing BDNF-AS exerts protective functions to increase the viability, inhibit the apoptosis and oxidative stress of Aβ_25-35-induced PC12 cells by negative regulation of BDNF.

Abbreviations: Aβ_25-35: amyloid beta peptide 25-35; AD: Alzheimer's disease; LncRNA BDNF-AS: long non-coding RNA brain-derived neurotrophic factor anti-sense; OS: Oxidative stress.

Keywords: Alzheimer’s disease; Aβ25-35; LncRNA BDNF-AS; PC12 cell; apoptosis; oxidative stress.

MeSH terms

Amyloid beta-Peptides / toxicity*
Animals
Apoptosis / physiology
Brain-Derived Neurotrophic Factor / genetics*
Brain-Derived Neurotrophic Factor / metabolism*
Cell Survival / physiology
Membrane Potential, Mitochondrial / physiology
Oxidative Stress / physiology
PC12 Cells
Peptide Fragments / toxicity*
RNA Interference*
RNA, Antisense / genetics*
RNA, Long Noncoding / genetics*
Rats
Reactive Oxygen Species / metabolism

Substances

Amyloid beta-Peptides
Brain-Derived Neurotrophic Factor
Peptide Fragments
RNA, Antisense
RNA, Long Noncoding
Reactive Oxygen Species
amyloid beta-protein (25-35)