HIF-1α/BNIP3/Notch1 Pathway Based Research On The Mechanism Of HSYA-Regulated Autophagy In Treating Ischemic Stroke

ObjectiveTo explore the mechanism by which HSYA affects autophagic cerebral infarction by regulating the HIF-1α/BNIP3/Notch1 pathway through molecular docking prediction and animal experiments.Objective to supply the premise for the clinical application of Hsya within the prevention and treatment of stroke.Methods1.The model was ready then the power of HYSA was evaluated in rats.The model of focal middle artery occlusion(MCAO)was established by suture occlusion in Sprague Dawley(SD)rats.The model was evaluated by the score of Zea longa.The m NSS score of medicine operate was evaluated in every cluster.Infarct area was calculated using TTC staining.Histopathological studies were performed using HE staining.Terahertz time-domain spectroscopy experiments were performed to detect infarcted tissue.2.The expression of autophagy LC3 template RNA and organic compound in rat hippocampus was detected by visible light quantitative PCR and Western blot3.Whether HSYA affects HIF-1α/BNIP3/Notch1 pathway proteins to interfere with autophagy was predicted by molecular docking technology.Docking scores were calculated and 3D maps of docking patterns were drawn to predict whether small molecule compounds have strong binding activity to protein targets.4.Fluorescence quantitative PCR and Western blot were accustomed observe the expression of HIF-1α,BNIP3,Notch1 ribonucleic acid and macromolecule in rat hippocampus.Results1.Compared with I / R group,the MNSs score of Hsya group was significantly different(P<0.05),with a decreasing trend.As time increased,the symptom scores in the I/R,HSYA and Edaravone groups all tended to improve,with the HSYA trend being the most pronounced.The results showed that Hsya may improve the systema nervosum injury in rats,and therefore the injury was improved with the treatment time.TTC staining results showed that HSYA cluster and edaravone cluster was considerably minimized(P<0.05).The infarction size of edaravone cluster was over that of Hsya cluster(P<0.05).This indicates that HSYA can be used to achieve a therapeutic effect by reducing the infarct area of brain tissue.The HE staining results showed Hsya cluster had additional regular cell structure and clearer cell level.this means that Hsya will improve the degree of cell injury and scale back cell injury.The results of rate time-domain spectrographic analysis show that the HSYA cluster has absorbed less of the specific frequency bands than the I/R group,indicating that HSYA had a significant therapeutic effect.2.RT-q PCR: The relative expression of LC3 ribonucleic acid in hippocampus of Hsya cluster was considerably less than that of I/R cluster(P<0.05).The decrease of LC3 macromolecule level suggests that autophagy is smothered,which indicates that Hsya can inhibit autophagy to achieve the therapeutic effect of the disease compared with I/R group after72 hours of molding.Western Blot: the expression of hippocampal LC3 supermolecule within the HSYA cluster was considerably reduced,and therefore the distinction was statistically important(P<0.05).The above results indicated that HSYA was able to inhibit autophagy and treat the disease once seventy two hours of molding,as shown by the reduced expression of autophagy issue LC3.3.A comparative analysis by comparing the scores of key targets and proligands indicated that HSYA had good binding activity to HIF-1α,BNIP3 and Notch1,to which it could bind tightly.However,the binding activity with LC3 was low and the binding was not tight.4.RT-q PCR: Compared with the I/R cluster,the expression of HIF-1α,BNIP3 and Notch1 macromolecule within the hippocampus of the HSYA cluster was considerably reduced(P<0.05).The results indicated that HSYA was able to scale back the expression of HIF-1α,BNIP3 and Notch1 72 hours after molding.Western Blot: The relative expression of HIF-1α,BNIP3 and Notch1 m RNA within the hippocampus of the HSYA cluster was considerably not up to that of the I/R cluster,and therefore the distinction was statistically important(P<0.05).The results indicated that HSYA might inhibit the expression of HIF-1α and BNIP3 and cut back the expression of Notch1,when seventy two hours of molding,it achieves the therapeutic effect of the disease.Conclusion1.Animal experiments demonstrated that HSYA can effectively treat the MCAO rat model by improving neurological deficits,reducing infarct size,improving cellular destruction and reducing the absorption of specific spectra.2.Animal experiments demonstrated that autophagy marker protein LC3 was inhibited in the HSYA-treated MCAO rat model 72 hours after molding,indicating that autophagy was inhibited.3.Molecular docking predicted that HSYA may affect autophagy via the HIF-1α/BNIP3/Notch1 pathway.4.Animal experiments demonstrated that 72 hours after molding,the HIF-1α/BNIP3/Notch1 pathway was inhibited in the HSYA-treated MCAO rat model,and the HIF-1α/BNIP3/Notch1 pathway was inhibited,and the trend of regulation of the pathway factors s showed a correlational statistics with the trend of autophagy regulation.this implies that HSYA may inhibit excessive autophagy by inhibiting the HIF-1α/BNIP3/Notch1 pathway to treat cerebral ischemia.In conclusion,after 72 hours of cerebral ischemia,HSYA may inhibit excessive autophagy by inhibiting the HIF-1α/BNIP3/Notch1 pathway to treat ischemic stroke.