This study was designed to investigate the effect of naringin in oxygen-glucose deprivation/reoxygenation (OGD/R) model and its mechanism. The target gene of naringin and the enriched pathways of the gene were searched and identified using bioinformatics analysis. Then OGD/R model was built using PC12 cells, after which the cells were treated with different concentrations of naringin. Subsequently, cell proliferation and apoptosis were evaluated by cell counting kit-8 (CCK-8) and flow cytometry assays, respectively. Meanwhile, the expression of NFKB1 in PC12 cells underwent OGD/R-induced injury was detected by qRT-PCR, while apoptosis-related and pathway-related proteins were checked by Western blot. DCF-DA kit was utilized to measure the level of ROS. Our results revealed that NFKB1, which was upregulated in MACO rats and OGD/R-treated PC12 cells, was a target gene of naringin. Naringin could alleviate OGD/R-induced injury via promoting the proliferation, and repressing the apoptosis of PC12 cells through regulating the expression of NFKB1 and apoptosis-associated proteins and ROS level. Besides, the depletion of NFKB1 was positive to cell proliferation but negative to cell apoptosis. Moreover, the depletion of NFKB1 enhanced the influences of naringin on cell proliferation and apoptosis as well as the expression of apoptosis-related proteins and ROS level. Western blotting indicated that both naringin treatment and depletion of NFKB1 could increase the expression of HIF-1α, p-AKT, and p-mTOR compared with OGD/R group. What’s more, treatment by naringin and si-NFKB1 together could significantly increase these effects. Nevertheless, the expression of AKT and mTOR among each group was almost not changed. In conclusion, naringin could prevent the OGD/R-induced injury in PC12 cells in vitro by targeting NFKB1 and regulating HIF-1α/AKT/mTOR-signaling pathway, which might provide novel ideas for the therapy of cerebral ischemia-reperfusion (I/R) injury.

本研究旨在研究柚皮苷在氧-葡萄糖剥夺/复氧（OGD/R）模型中的作用及其机制。通过生物信息学分析，对柚皮苷的靶基因及其富集途径进行了搜索和鉴定。然后使用PC12细胞建立OGD/R模型，然后用不同浓度的柚皮苷处理细胞。随后，分别通过细胞计数试剂盒-8 (CCK-8) 和流式细胞术检测评估细胞增殖和凋亡。同时，通过qRT-PCR检测受OGD/R诱导损伤的PC12细胞中NFKB1的表达，通过Western blot检测凋亡相关和通路相关蛋白。DCF-DA试剂盒用于测量ROS水平。我们的结果显示，在 MACO 大鼠和 OGD/R 处理的 PC12 细胞中上调的 NFKB1，是柚皮苷的靶基因。柚皮苷可以通过促进PC12细胞增殖、调节NFKB1和凋亡相关蛋白的表达和ROS水平来抑制PC12细胞的凋亡，从而减轻OGD/R诱导的损伤。此外，NFKB1的消耗对细胞增殖呈阳性但对细胞凋亡呈阴性。此外，NFKB1的缺失增强了柚皮苷对细胞增殖和凋亡的影响，以及凋亡相关蛋白的表达和ROS水平。Western印迹表明，与OGD/R组相比，柚皮苷处理和NFKB1的消耗均能增加HIF-1α、p-AKT和p-mTOR的表达。更重要的是，柚皮苷和 si-NFKB1 一起治疗可以显着增加这些效果。尽管如此，各组中AKT和mTOR的表达几乎没有变化。总之，柚皮苷可以通过靶向NFKB1和调节HIF-1α/AKT/mTOR信号通路来预防OGD/R诱导的PC12细胞体外损伤，这可能为脑缺血再灌注的治疗提供新的思路（I/ R) 伤害。