Effect of mitochondrial reactive oxygen species inhibitor EUK-134 on mitochondrial associated proteins PGC-1α and VDAC1 in rats with cerebral ischemia-reperfusion injury

doi:10.3969/j.issn.1006-7795.2024.06.011

Abstract

Abstract: Objective To investigate the effects of Eukarion-134 (EUK-134), a scavenger of reactive oxygen species (ROS), on peroxisome proliferator activated receptor gamma coactivator-1α (PGC-1α) and mitochondrial voltage-dependent anion channel 1 (VDAC1) in middle cerebral artery occlusion (MCAO) and reperfusion rats, and to explore the impact of ROS generation on mitochondrial structure and function after cerebral ischemia. Methods Healthy male Sprague Dawley (SD) rats were divided into three groups with a random number table method: Sham group, MCAO group, and MCAO+EUK-134 (EUK-134) group, with 6 rats in each group. A model of MCAO was induced with the intraluminal suture method. The rats underwent 90 min of MCAO and then were reperfused for 24 h by withdrawal of the filament. The heart rate, mean arterial pressure, and temperature were maintained in the normal range during surgery. EUK-134(10 mg/kg) was injected subcutaneously in rats of the EUK-134 group, and rats in the sham and MCAO groups received equal volumes of solvent. The expression of PGC-1α in ischemic brain tissue was assessed by Western blotting. Immunofluorescence staining was applied to detect the expression of VDAC1 in the ischemic penumbra of rats. The ROS positive cells were labeled with an oxidant-sensing fluorescent probe (2′,7′-dichlorofluorescin diacetate, H2DCF-DA) staining, and double staining for VDAC1 and ROS were performed. Results ① The expression of PGC-1α protein level in the ischemic brain tissue of rats in the MCAO group after 24 h reperfusion was significantly lower than that of the sham group (P<0.05). The PGC-1α protein level in the ischemic brain tissue of the EUK-134 group was significantly higher than that of the MCAO group (P<0.05). ② After 24 h reperfusion, the number of VDAC1 positive cells in the ischemic penumbra of the MCAO group increased significantly compared to the sham group (P<0.05), while the number of VDAC1 positive cells in the ischemic penumbra of the EUK-134 group was significantly lower than that of the MCAO group (P<0.05). VDAC1 positive cells were colocalized with the neuronal marker NeuN in the brain ischemic penumbra. ③ In the MCAO group, VDAC1 positive cells were colocalized with ROS in the ischemic penumbra, and the number of VDAC1/ROS double positive cells was considerably increased compared to that of the sham group (P<0.05). The number of VDAC1/ROS double positive cells in the penumbra of EUK-134 treated rats decreased significantly compared to the MCAO group (P<0.05). Conclusions The ROS scavenger EUK-134 could maintains mitochondrial function by affecting the expression of the mitochondrial biogenic regulator PGC-1α and the substance transport channel VDAC1, thereby ameliorating cerebral ischemia-reperfusion injury.

Key words: cerebral ischemia/reperfusion, reactive oxygen species, EUK-134, PGC-1α, VDAC1

CLC Number:

R743.3

Li Yakun, Li Wei, Yang Xueqi, Shi Wenjuan, Zhu Yuequan, Zhao Yongmei. Effect of mitochondrial reactive oxygen species inhibitor EUK-134 on mitochondrial associated proteins PGC-1α and VDAC1 in rats with cerebral ischemia-reperfusion injury[J]. Journal of Capital Medical University, 2024, 45(6): 1016-1022.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2024.06.011

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2024/V45/I6/1016

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