ABSTRACT
Stroke is a major public health problem with an imperative need for a more effective and tolerated therapy. Neuroprotective therapy may be an effective therapeutic intervention for stroke. The morbidity and mortality of stroke-induced secondary brain injury is mainly caused by neuronal apoptosis, which can be executed in a caspase-dependent or apoptosis inducing factor (AIF)-dependent manner. As apoptosis is an energy-dependent process with a relative time delay, abnormal energy metabolism could be a significant and fundamental pathophysiological basis of stroke. To our knowledge, convincible evidences that AMPK inhibition exerts neuroprotection in cerebral ischemia injury via anti-apoptosis remain to be investigated. Accordingly, the aims of this study were to investigate the protective effects of AMPK inhibitor BML-275 on cerebral ischemic/reperfusion (I/R) injury and to elucidate the underlying mechanisms. Cerebral ischemia was induced by transient middle cerebral artery occlusion (tMCAO) in male C57BL/6 mice. The therapeutic effects of BML-275 were evaluated by infarct sizes, neurological scores and the proportion of apoptotic neurons after 24 h of reperfusion. The cell apoptosis markers cyt c and AIF were also evaluated. The results showed that intraperitoneally administration of BML-275 alleviate the cerebral infarction, neurological deficit and neuronal apoptosis induced by MCAO. BML-275 simultaneously induces anti-apoptosis and decreases the expression of cyt c and AIF. This study supports the hypothesis that anti-apoptosis is one of potential neuroprotective strategies for the treatment of stroke.
Subject(s)
AMP-Activated Protein Kinases/antagonists & inhibitors , Apoptosis Inducing Factor/antagonists & inhibitors , Brain Ischemia/drug therapy , Cytochromes c/antagonists & inhibitors , Neuroprotective Agents/pharmacology , Pyrazoles/pharmacology , Pyrimidines/pharmacology , AMP-Activated Protein Kinases/metabolism , Animals , Apoptosis/drug effects , Apoptosis Inducing Factor/genetics , Apoptosis Inducing Factor/metabolism , Brain Ischemia/metabolism , Brain Ischemia/pathology , Cytochromes c/genetics , Cytochromes c/metabolism , Dose-Response Relationship, Drug , Male , Mice , Mice, Inbred C57BL , Molecular Structure , Neurons/drug effects , Neurons/metabolism , Neurons/pathology , Neuroprotective Agents/chemical synthesis , Neuroprotective Agents/chemistry , Pyrazoles/chemical synthesis , Pyrazoles/chemistry , Pyrimidines/chemical synthesis , Pyrimidines/chemistry , Structure-Activity RelationshipABSTRACT
This study aimed to investigate the expression, function, and possible mechanism of Src in the Hep-2 cell line. We used Src-specific small interfering RNA (siRNA) to inhibit the expression of Src in Hep-2 cells. RT-PCR and Western blot were applied to evaluate the expression level of Src after RNA interference, and the MTT assay and flow cytometry were used to observe the expression of PI-3 K and Akt. siRNA can downregulate the expression of Src in Hep-2 cells. Downregulation of Src decreased PI-3 K and Akt expression. We found that Src knockdown inhibits the proliferation of Hep-2 cells and the growth of laryngeal carcinoma in vivo. This study has demonstrated that Src participates in the regulation of apoptosis through the Src/PI-3 K/Akt signaling pathway in the Hep-2 cell line. Silencing of Src by siRNA is a viable approach in laryngeal carcinoma treatment.
