Protective Effects of UCF-101 on Cerebral Ischemia-Reperfusion (CIR) is Depended on the MAPK/p38/ERK Signaling Pathway

Abstract

This study was aimed to investigate the treatment mechanisms of 5-[5-(2-nitrophenyl) furfuryliodine]-1,3-diphenyl-2-thiobarbituric acid (UCF-101) in cerebral ischemia-reperfusion (CIR) model rats. Total of 54 healthy male Wistar rats were randomly assigned into three groups, namely sham group, vehicle group, and UCF-101 group. The CIR-injured model was established by right middle cerebral artery occlusion and reperfusion. Neurological function was assessed by an investigator according to the Longa neurologic deficit scores. Meanwhile, the cerebral tissue morphology and apoptotic neurons were evaluated by H&E and TUNEL staining, respectively. Additionally, the expressions of caspase 3, p-p38, and p-ERK were detected by immunohistochemistry or/and Western blotting assays. As results, neurologic deficit and pathological damage were obviously enhanced and TUNEL positive neurons were significantly increased in CIR-injured rats, as compared with those in sham group. Furthermore, the expressions of caspase 3, p-p38, and p-ERK were also significantly increased in vehicle group than those in sham group (P < 0.05). However, UCF-101 treatment could markedly weaken the neurologic deficit with lower scores and improve pathological condition. After UCF-101 treatment, TUNEL positive neurons as well as the expression of caspase 3 were significantly decreased than those in vehicle group (P < 0.05). Besides, p-p38 was decreased while p-ERK was increased in UCF-101 group than those in vehicle group (P < 0.05). Therefore, we concluded that the protective effects of UCF-101 might be associated with apoptosis process and MAPK signaling pathway in the CIR-injured model.

Keywords: Apoptosis neurons; Cerebral ischemia–reperfusion; MAPK; UCF-101.

Fig. 1

UCF-101 alleviated cerebral ischemia–reperfusion-induced neuronal morphological changes. Hematoxylin and eosin (H&E) staining was conducted to observe the cerebral tissue morphology at 12 h after reperfusion (×200)

Fig. 2

UCF-101 reduced apoptosis in cerebral ischemia–reperfusion rats. a Apoptotic neurons in cortex by TUNEL assay (×200); b The expressions of caspase 3 by immunohistochemistry (×200)

Fig. 3

The expression of p-p38MARK by immunohistochemistry (×200)

Fig. 4

The expressions of p-p38 and p-ERK were changed after UCF-101 treatment. The expression of p-p38 by Western blotting (a) and the relative protein expression (b). The expression of p-ERK by Western blotting (c) and the relative protein expression (d). Columns mean (n = 3); bars SD; **P < 0.01 versus vehicle group