miR‑429 and GATA4 may participate in cerebral ischemic stroke by regulating autophagy and apoptosis: the effect of Chlorogenic acid

Abstract
Autophagy is a double-edged sword for maintaining neural system homeostasis during development of cerebral ischemia. However, the potential molecular mechanisms behind that remain unclear. This study investigated the miR-429 and its target GATA4 alterations, autophagy mediators and apoptosis in ischemic stroke alone and also in combination with chlorogenic acid (CGA).
Male Wistar rats were assigned into three groups (n=8): sham, IR (ischemia-reperfusion, ischemia-reperfusion, transient cerebral ischemia inducing by common carotid artery occlusion and reperfusion), IR+CGA (30 mg/kg, ip; intraperitoneally, 10 minutes prior to ischemia and 10 minutes before reperfusion.). miR-429, GATA4, c-Caspase-3/p-Caspase-3 ratio, LC3-I, LC3-II, Beclin1 and p62 measured by Real time PCR and also Western blot assays. At the end of the experiment, we observed increased miR-429 gene expression (P<0.05) and c-Caspase-3/p-Caspase-3 ratio (P<0.01) as well as decreased GATA4 protein expression (P<0.001) in IR group. In addition, autophagy was over-activated markedly in the brain of CCAO rats as evidenced by an increased ratio of LC3-II/I and Beclin1 protein expression and decreased expression of p62 after 24 h of reperfusion (P<0.001). Moreover, immunohistochemistry studies found that cerebral IR significantly increased the ratio of total LC3 immunoreactivity in the cortex tissue of male rats (P<0.001). Treatment with CGA significantly attenuated autophagic activity as well as apoptosis and reversed aforementioned molecule levels. Taken together, these results suggested that ischemic insult can increase autophagic activities and apoptosis possibly by miR‑429 and GATA4 alterations in the brain cortex after cerebral IR insult which can be alleviated by CGA as a potential therapy for someone afflicted with ischemia.