UPLC-Q-TOF/MS-Based Metabonomics Study On The Anti-ischemic Stroke Effects Of Wogonoside

Stroke is the leading cause of disability and death,with ischemic strokes accounting for80%of all cases.Due to the blockage of blood clot,arterial blood flow is decreased and brain blood supply to is inadequate,followed by an ischemic cascade reactions and ischaemic damage to brain tissues.The brain damage was further exacerbated by restoration of cerebral blood flow after reperfusion,known as ischaemic reperfusion injury,inducing oxidative stress,leading to lipid peroxidation and DNA damage.Stroke impact is global and there are few treatment options available.The only currently approved drug for acute ischemic stroke is recombinant thrombolytic tissue plasminogen activator（rt PA）,but due to strict eligibility criteria and narrow time window for treatment,only a very small number of patients can be treated with rt PA.Therefore,the development of neuroprotective drugs that protect brain cells from both ischemia/reperfusion（I/R）injury.Scutellaria baicalensis has a long history of medicinal use in China and is one of the most commonly used herbs in clinical practice.Modern pharmacological studies have shown that wogonoside,the main active ingredient of Scutellaria baicalensis,has a wide range of pharmacological effects,including antioxidant,anti-inflammatory and anti-tumour.It has been reported in the literature that wogonoside can significantly alleviate H₂O₂-induced cell damage in PC12 cells,however,no data have been reported on how wogonoside exerts its protective effect and whether the effect persists in vivo.Metabolomics examines changes in the metabolites of biological systems,including tissues,cells,body fluids or organisms,when they are stimulated or perturbed（by drug treatment or environmental changes）.Metabolomic approach integrated with common biological methods hold great promise to elucidate the complex pathogenesis of ischaemic stroke.An in vitro model of oxygen glucose deprivation/reperfusion（OGD/R）model was constructed and LC MS metabolomics was used to investigate the endogenous metabolite changes in PC12 cells after OGD/R induction and wogonoside intervention,and to explore the potential mechanisms underlying the protective effects of wogonoside.The results suggested that wogonoside may exert neuroprotective effects by reducing oxidative stress,enhancing anti-inflammatory capacity,improving lipid metabolism and amino acid metabolism.OGD/R treatment induced excessive production of mitochondrial ROS,reduced mitochondrial membrane potential level and increased the apoptosis of PC12 cells.The neuroprotective effect of wogonoside was inhibited by Nrf2 and Sirt3 inhibitors,suggesting that wogonoside may enhance antioxidant enzyme activity,improve mitochondrial function and partially attenuate apoptosis through activation of the Nrf2/Sirt3 pathway.Furthermore,an I/R model of middle cerebral artery occlusion（MCAO）in SD rats was used to characterize the neuroprotective effects of wogonoside in vivo.Wogonoside markedly decreased mortality,neurological deficit score,cerebral infarct size and brain water content of MCAO rats,ameliorated I/R induced oxidative stress injury.Our study provides new data to support the further clinical adoption and application of wogonoside.