Effect Of Bujingyishipian On Retinal Pigment Epithelium In Myopia Mice Based On Single-cell RNA Sequencing Technology

Research ObjectiveTo explore the traditional Chinese medicine bujingyishi tablet to improve the gene function of retinal pigment epithelial cells in myopia mice,and to provide a theoretical basis for early gastric myopia in bujingyishi tablet.Research MethodNormally healthy mice were randomly assigned into negative control group,myopia model group and Chinese medicine gavage(bujingyishi tablet suspension gavage)group.Both eyes of the negative control group did not need to be covered.Both myopia model group and Chinese medicine gavage group were used.Cover the right eye for shape deprivation.After the experiment is performed for 3 weeks,the model is confirmed to be successful.Then the Chinese medicine gavage group is administered with the traditional Chinese medicine bujingyishi tablet suspension for 3 weeks.The eye axis is measured before and after the experiment.The right eye was taken as a single cell to capture retinal RPE cells for RNA sequencing,and functional analysis was performed using gene set enrichment analysis and Kyoto gene and genome encyclopedia pathway analysis.Research Result1.A total of 3900 genes were identified in the myopia model group compared with the negative control group.The difference analysis revealed 617 genes,96 up-regulated genes and 521 down-regulated genes.The GO analysis corresponds to 1801 GO terms and 1039 GO terms with statistical significance,of which 28 are related to oxidative stress,and related GO terms such as circadian rhythm,epithelial apoptosis,vision,and neurons and neurotransmitters.There are 60 GO items and 61 protein related GO items.KEGG pathway analysis detected a total of 219 pathways and 87 statistically significant pathways.Among them,oxidative phosphorylation-related pathways such as glycolysis/gluconeogenesis,c AMP pathway,Notch pathway,dopaminergic pathway,and GABA ergic pathway were found.,Glutamic acid,alanine,aspartic acid,oxytocin pathway and other neurotransmitter-related pathways,Alzheimer's disease pathway,AMPK signaling pathway,m TOR signaling pathway and other pathways related to neurodegenerative diseases;2.Compared with the myopia model,a total of 1945 genes were identified in the Chinese medicine gavage group.Difference analysis revealed 170 genes,52 up-regulated genes and 118 down-regulated genes.GO analysis corresponds to 785 GO items,745 GO items with statistical significance,of which 17 are related to oxidative phosphorylation,36 are GO related to nervous system development,and 13 are GO related to protein regulation.The related GO term was found to positively regulate epithelial cell proliferation and negatively regulate cell death.KEGG pathway analysis detected a total of 163 pathways,of which 50 were statistically significant.Among them,PI3K-Akt signaling pathway,Wnt signaling pathway,Hedgehog signaling pathway and other oxidation-related pathways were found.Parkinson's disease,Huntington's disease,muscle Pathogenesis related to neurodegenerative diseases such as cord sclerosis,and pathways related to endoplasmic reticulum stress and autophagy such as protein processing in the endoplasmic reticulum and AMPK signaling pathway;3.A total of 21 differential genes were analyzed in both groups.We found that Insig1 and Scd1 are related to endoplasmic reticulum stress,the apoptosis marker protein Caspase substrate Pabpc1,and the acute cell phosphorylation apoptosis-related protein PSMD11.In the myopia model group,the expression of the myopia model group was significantly up-regulated,and in the Chinese medicine gavage group,the expression of the myopia model group was significantly down-regulated(P <0.05).Research Conclusion1.RPE cell dysfunction appears in the development of myopia,which is related to the expression of related genes and the regulation of pathways of glycolysis,neurodegenerative diseases,protein regulatory functions.2.Bujing Yishi tablet can slow down the oxidative stress of retinal RPE cells in myopic mice,balance endoplasmic reticulum stress and autophagy,protect the function of RPE cells,and maintain the normal activity of photoreceptor cells.