Study On The Effects And Mechanism Of Gastrodia Elata Extract On Hypoxia And Hypoglycemia Injury In PC12 Cell

To date,stroke has become one of the most serious threats to public health in the21 st century,and is the second leading cause of death and third leading cause of disability worldwide,with ischemic stroke predominant.Stroke mainly causes neuronal damage.As a traditional Chinese medicinal material,Gastrodia elata is mainly used for the treatment of dizziness,insomnia,epilepsy,neurasthenia and other diseases.This study provides some theoretical guidance for the study of the mechanism of ischemic stroke,and also provides an in-depth discussion of the active components and neural mechanism of Gastrodia elata,and provides important insights for the network pharmacological analysis of Gastrodia elata.Through network pharmacology screening analysis,a total of 72 proteins may be the targets of Gastrodia elata in anti-ischemic stroke.GO and KEGG enrichment analysis indicated that 50 potential targets were mainly mapped to the MAPK signaling pathway.Further molecular docking results showed that the core compound of Gastrodia elata binds the most closely to the targets MAPK10 and PTGS1,and the main pathway is enriched to the MAPK signaling pathway.In this experiment,the hypoxia and hypoglycemia of cells was mainly used to simulate the in vitro ischemic stroke model.Through the activity detection of crude extracts of different polarities of Gastrodia elata,it was found that the methanol extract of Gastrodia elata had a good effect on the repair of hypoxia and glucose damage in PC12 cells.The best drug effect was when the drug concentration was0.005 μg/μL.Based on this,we further explored the damage repair and action mechanism of the active ingredients gastrodin and balisein on the hypoxia and glucose deprivation model of PC12 cells.The results showed that both gastrodin and balisonoside could reduce the release of intracellular reactive oxygen species(ROS)and promote cell proliferation to a certain extent,indicating that both have neurological repair effects on PC12 cells stimulated by hypoxia and glucose..In order to further explore the mechanism and pathway of the two,combined with the previous network pharmacology analysis results,the protein activity analysis of gastrodin and balisonoside and the main targets in the MAPK signaling pathway was carried out.Through protein detection,it was found that balisonoside can reduce the phosphorylation level of MAPK10 and promote the phosphorylation of P38 protein.MAPK10 is mainly related to neuronal apoptosis,and the increase of the phosphorylation level of P38 protein is mainly related to cell proliferation.It regulates the phosphorylation level through two targets to inhibit apoptosis,reduce LDH release and the production of intracellular reactive oxygen species,thereby playing a neuroprotective role.Gastrodin has nerve repair activity,but has no significant effect on MAPK10 and P38 protein phosphorylation levels,indicating that gastrodin does not act through MAPK signaling pathway,but through other signaling pathways,which is in line with the network pharmacology analysis result.