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

OBJECTIVE

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

METHODS

PC12 cells were induced by Aβ 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 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-induced PC12 cells, with the elevated cell viability. Moreover, silencing BDNF-AS inhibited the apoptosis of Aβ-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-induced PC12 cells by negative regulation of BDNF.

ABBREVIATIONS

Aβ: 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.