| Objective:1.To discuss the relationship between deficiency of kidney essence and ischemic stroke from the theory of "kidney stores essence and marrow generates brain",to further elaborate the effect and intervention mechanism of Radix rehmanniae and Cornus officinalis in the treatment of CIS,and to summarize the role of energy metabolism disorders in the pathogenesis of ischemic stroke and its relationship with the mechanism of "deficiency of kidney essence".We also summarized the role of energy metabolism disorders in the pathogenesis of ischemic stroke and its relationship with the mechanism of "kidney essence deficiency".2.To detect the effect of the combination of Radix rehmanniae and Cornus officinalis on the protein expression pattern of brain tissue of rats with cerebral ischemia,and to analyze the possible targets for the treatment of ischemic stroke with bioinformatics analysis,so as to provide direction for further mechanism research.3.To investigate the protective effect of the drug-containing serum of Radix rehmanniae-Cornus officinalis on SH-SY5 Y cells of OGD/R model and the effect on Sirt1/LKB1/AMPK energy metabolism signaling pathway of these cells,in order to explore the mechanism of this drug for the treatment of ischemic stroke.Methods:1.Starting from the literature on " kidney brain related " theory of traditional Chinese medicine(TCM)and stroke disease,we sort out the theoretical lineage and modern research on " curing stroke from kidney ",systemically evaluate the clinical efficacy of "" supplementing kidney and filling lean pulp "" method in the treatment of ischemic stroke,and review the literature of multiple studies on the effect of energy metabolism disorders and their intervention mechanisms in the process of cerebral ischemic injury,To explore the biological basis of " supplementing the kidney to fill the spermatozoon " method to treat ischemic stroke by improving energy metabolism.2.To establish ischemic stroke model using MCAO method on SD rats,and set up sham operation group,model group and treatment group,rats in each group were treated accordingly,sham operation group underwent sham operation without plug line,treatment group was gavaged with Radix rehmanniae-Cornus officinalis,model group was gavaged with equal amount of distilled water,after continuous intervention for 7 days,brain tissues of infarcted hemisphere were taken for TMT proteomic detection,In addition,the peptide information obtained from the assay was subjected to differential protein screening and further raw signal analysis to summarize the biological processes,structural localization,and signaling pathways of the drug for intervention on the infarcted brain tissue.3.SH-SY5 Y cells were subjected to OGD 4h / R 24 h modeling to prepare the drug containing serum of blank serum and Radix rehmanniae-Cornus officinalis in advance,the blank,control,blank serum,drug containing serum and selisistat(SIRT1 inhibitor)groups were set up,and after intervention,cell morphology changes,cell viability,ATP / AMP content changes and protein and m RNA expression of each factor of SIRT1 / LKB1 / AMPK signaling pathway were observed,To investigate the protective effects and regulation of energy metabolism pathways of Radix rehmanniae-Cornus officinalis on an in vitro model of ischemic stroke.Results:1.Through classical literature and modern research literature,we found that "kidneybrain syncretism" is an important pathogenesis of ischemic stroke,and "tonifying the kidney and filling the essence" proposed by the theory of kidney-brain generation is an important treatment method,and Meta-analysis evaluated that this treatment method has achieved good clinical efficacy.A number of modern studies have confirmed that energy metabolism is the initiating link in the pathogenesis of ischemic stroke,and intervention in this pathophysiological process can effectively block the ischemic cascade reflection and reduce brain tissue damage.2.The proteomic study found that there were 80 differentially expressed proteins in the brain tissues of rats before and after modeling,of which 41 were up-regulated and 39 were down-regulated,and the functions of these proteins were mainly focused on the regulation of the activities of various enzymes,such as oxidoreductase,phosphotransferase,protein phosphatase,hydrolytic enzymes,etc.After the application of Radix rehmanniae-Cornus officinalis intervention,the differentially expressed proteins between them and the model group were 993,of which 756 were up-regulated.993,of which 756 were up-regulated and237 were down-regulated,and these proteins increased the activity of multiple molecule binding.The differential proteins were involved in signaling pathways focused on ketone body synthesis and degradation,glutamatergic synapses,circadian rhythms,long-term potentiation,phosphatidylinositol signaling system,2-oxocarboxylic acid metabolism,Gn RH secretion,cholinergic synapses,amphetamine addiction and dopaminergic synapses.Among these targets of intervention,there were significant changes in the factors associated with the Sirt1/LKB1/AMPK energy metabolism pathway.This trend was also reflected in the hierarchical clustering analysis,KEGG enrichment and subcellular localization analysis,reflecting the importance of changes in energy metabolic processes before and after modeling and before and after drug intervention.3.The morphology of SH-SY5 Y cells damaged by glucose and oxygen deficiency was significantly impaired,and cell viability and ATP production were reduced.After the intervention of each group,the drug-containing serum increased the energy supply to the cells,restored the impaired cell viability and increased the expression of various factors of Sirt1/LKB1/AMPK signaling pathway,while the control and blank serum groups did not have this effect.In contrast,after inhibition of Sirt1 activity with Selisistat,the expression of all factors of the Sirt1/LKB1/AMPK signaling pathway decreased in the drug-containing serum+inhibitor group,and the cellular energy supply was more severely impaired than that of the drug-containing serum group,with reduced cell morphology and ATP supply.Conclusion:1.Tonifying the kidney and replenishing essence is an effective method for the treatment of ischemic stroke.Radix rehmanniae-Cornus officinalis is the representative prescription of this treatment,and the disorder of energy metabolism is an important pathogenesis of this disease.Exploring the relationship between the effect of this drug on and the function of energy metabolism is helpful to understand the connotation of Tonifying the kidney and replenishing essence.2.There were significant differences in brain protein expression between Radix rehmanniae-Cornus officinalis treatment group and MCAO model group.The expression of energy metabolism signal pathway related factors such as LKB1,Ca MKK family and AMPK in the treatment group was higher than that in the model group,which provided direction for further study of the intervention effect of drugs on ischemic stroke.3.Radix rehmanniae-Cornus officinalis medicinal serum can significantly restore the nerve cell injury caused by glucose deficiency and hypoxia,while inhibiting SIRT1 / LKB1 /AMPK signal pathway can weaken its effect,suggesting that improving energy metabolism function is one of the possible mechanisms of Radix rehmanniae-Cornus officinalis in the treatment of cerebral ischemic injury. |
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