Study on protective mechanism of kaempferol against cerebral ischemia/reperfusion injury in vitro and in vivo / 中国药房

ABSTRACT

OBJECTIVE To explore the protective mechanism of kaempferol （KAE）on cerebral ischemia/reperfusion （CI/R） injury in vitro and in vivo . METHODS Firstly，the potential targets of KAE in the treatment of CI/R injury were preliminarily screened by network pharmacology. AutoDock Vina software was used to conduct molecular docking between KAE and the top 10 core targets ，and the binding affinity was used as the evaluation standard to further clarify the possible mechanism of KAE in treating CI/R injury. Finally ，the above results were verified by in vivo and in vitro experiments. The oxygen glucose deprivation/ reperfusion（OGD/R）HT22 cell injury model and the middle cerebral artery occlusion （MCAO）rat model were constructed. The cell activity was detected by CCK- 8 method. The neural function score and TTC staining were performed on the rats and their brain tissues. The phosphorylation levels of protein kinase B （Akt）and Src in HT 22 cells and brain tissue of rats were detected by Western blot. RESULTS The results of network pharmacology screening showed that KAE in the treatment of CI/R injury was closely related to 10 core targets including prostaglandin-endoperoxide synthase 2，matrix metalloproteinase 9，JUN，Akt1，tumor necrosis factor ，caspase-3，mitogen activated protein kinase 8，intercellular cell adhesion molecule 1，vascular cell adhesion molecule 1 and Src. The results of molecular docking showed that KAE was stably bound with Akt 1 and Src . The results of in vitro and in vivo experiments showed that KAE could significantly improve the survival rate of OGD/R-injuried HT 22 cells （P＜0.05）， significantly reduced the neurological function score of MCAO model rats （P＜0.05），significantly reduces the volume of cerebral infarction in rats （P＜0.05），and significantly increased the phosphorylation levels of Akt and Src in HT 22 cells and brain tissue of rats（P＜0.05），which showed a dose dependent trend. CONCLUSIONS KAE may play a neuroprotective role by regulating the phosphorylation expression of Akt and Src ，thus treating CI/R injury.