Influence of lncRNA ANRIL on neuronal apoptosis in rats with cerebral infarction by regulating the NF-Œ∫B signaling pathway
Authors: J-H Zhao
B Wang
C-W Xu
J-R Wang
X-H Wang
Journal: European review for medical and pharmacological sciences (2019)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: lncRNA ANRIL; neuronal apoptosis; cerebral infarction; NF-Œ∫B signaling pathway; rats
Affiliations: Department of Neurology, First Affiliated Hospital of China Medical University, Shenyang, China. .

Abstract:  Objective: The aim of this study was to evaluate the influence of long non-coding ribonucleic acid (lncRNA) antisense non-coding RNA in the INK4 locus (ANRIL) on neuronal apoptosis in rats with cerebral infarction (CI), and to further explore the underlying mechanism of lncRNA ANRIL in the occurrence and development of CI. Materials and methods: A total of 60 adult male Wistar rats were randomly divided into three groups using a random number table, including sham group (n=20), CI group (n=20) and CI + lncRNA ANRIL knockdown group [CI + lncRNA ANRIL small-interfering RNA (siRNA) group, n=20]. Focal CI was constructed by suture occlusion. After successful modeling, lncRNA ANRIL siRNA was stereotactically injected into the lateral ventricle of the rats. 24 h after operation, the neurological function of the rats in each group was evaluated by the modified neurological severity score (mNSS). Meanwhile, the infarction area of brain tissues was evaluated using the triphenyl tetrazolium chloride (TTC) method. The protein expression levels of apoptosis-related genes, including B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax), were detected via Western blotting. Subsequently, immunofluorescence staining was performed to detect the expression and location of Caspase-3 in brain tissues. Moreover, the apoptosis level of rats in each group was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Furthermore, the expressions of nuclear factor-Œ∫B (NF-Œ∫B) signaling pathway-related proteins were detected via Western blotting. Results: Polymerase Chain Reaction (PCR) results revealed that the expression level of lncRNA ANRIL in the CI group was significantly increased when compared with that of the sham group (p&lt;0.05). The results of mNSS and TTC staining manifested that knockdown of lncRNA ANRIL could significantly reduce CI-induced neurological deficits and CI area (p&lt;0.05). At the same time, knockdown of lncRNA ANRIL markedly decreased the level of Bax, whereas increased the expression of Bcl-2 (p&lt;0.05). Besides, the number of apoptotic cells in the CI + lncRNA ANRIL siRNA group was remarkably decreased (p&lt;0.05). In addition, lncRNA ANRIL down-regulation remarkably inhibited the phosphorylation of p65 (p&lt;0.05). Conclusions: The inhibitory effect of lncRNA ANRIL knockdown on neuronal apoptosis in CI rats may be probably related to its inhibition of the NF-Œ∫B signaling pathway. Furthermore, lncRNA ANRIL inhibitor is expected to become a targeted drug in the clinical treatment of CI. 

Retracted: No

PubPeer comments:
 *[below]* Fig 3.  Influence of lncRNA ANRIL knockdown on hippocampus morphology in CI rats . After a 180-deg rotation, the CI panel matches the miR-431+siRNARho panel from *[above]* Fig 3 of [Han et al (2018)] . Could the authors check?  , 
 *[above]* Fig 3.  Influence of lncRNA ANRIL knockdown on hippocampus morphology in CI rats (magnification: 40u00d7). Sham: Sham operation group, CI: CI group, CI+ANRIL siRNA: lncRNA ANRIL knockdown group . *[below]* Fig 4 from [Li et al (2017)] .  Figure 4. Nissl staining of hippocampal neurons in the CA1 area in the four groups (200u00d7) .  --------------------------- *[above]* Fig 4.  Influence of lncRNA ANRIL knockdown on neuronal apoptosis in hippocampus of CI rats (magnification: 40u00d7). Sham: Sham operation group, CI: CI group, CI+ANRIL siRNA: lncRNA ANRIL knockdown group . *[below]* Fig 6 from Li et al (2017).  The cell apoptosis of brain neuronal cells of rats in the four groups by TUNEL staining .  , 
 #2  Another sighting! Fig 5 of [Han et al (2018)] .  Apoptosis rate of neurons in the hippocampus of rats from the normal, sham and model groups detected by TUNEL staining .  Could the authors comment on the disappearance of cells from that earlier version? ,