Gene Verification Of Diabetic Retinopathy And Analysis Of TangWangFang Signaling Pathway Based On Network Pharmacology

BackgroundDiabetes mellitus(DM)is a common endocrine disease characterized by disorders of glucose metabolism.The prevalence of diabetes and the number of diabetic patients are increasing rapidly.The International Diabetes Federation(IDF)estimates that by 2040,one in 10 adults with diabetes(20-79 years old)will be diagnosed with diabetes.By 2045.629 million diabetes patients will be diagnosed worldwide.The mortality rate of diabetes will be higher than that of AIDS,tuberculosis and malaria combined.Every year,about 5 million people die of diabetes.One diabetic died in six seconds.According to statistics,the total expenditure on health care for diabetes mellitus is expected to increase to US$802 billion by 2040.According to the Guidelines for Type 2 Diabetes in China,the prevalence of diabetes mellitus in adults in China is 10.9%.the incidence tends to be younger,the prevalence under 40 is as high as 5.9%.and the proportion of undiagnosed diabetes mellitus is higher.Diabetic retinopathy(DR)is one of the most common microvascular complications of diabetes mellitus.In China,the prevalence of retinopathy is 24.7%-37.5%in the diabetic population.The prevalence of DR is 50%-70%in diabetic patients with a course of 10-15 years.About 2%of the diabetic patients end up with diabetic retinopathy 15 years later.Total blindness and diabetic retinopathy are the main causes of vision loss between the ages of 20 and 74.However,less than 20%of diabetic retinopathy patients have been diagnosed at an early stage in China.The occurrence and development of diabetic retinopathy has greatly affected people's lives and quality of life.The course of diabetes,blood sugar level,blood pressure,blood lipid and other clinical factors have a certain impact on the occurrence and development of diabetic retinopathy.However,there are great individual differences in the course and severity of diabetic retinopathy.Some patients with long duration of diabetes have not developed diabetic retinopathy despite poor blood sugar control,while some patients develop diabetic retinopathy rapidly.This difference may be due to genetic polymorphism.In recent years,more and more people have known about the safety and biological distribution of gene delivery vectors.It has been proved that stem cells and gene therapy can reverse degenerative retinopathy.Drugs involve atrophic or damaged retinal tissue regeneration,neurotrophic factors,immune regulation,and new therapies for replacement of mutant genes.With the progress of science and technology,the number of genes that can be selected for intervention is also increasing.At present,the research on gene therapy for diabetic retinopathy is mostly focused on the use of related gene drugs against pigmented epithelium-derived factors,angiostatin.endostatin and vascular endothelial growth factor to inhibit the proliferation of endothelial cells and the formation of new blood vessels,so as to achieve the clinical purpose of preventing and treating diabetic retinopathy.At present.the pathogenesis and related genes of diabetic retinopathy still need to be further explored.Previous studies of our group have shown that compared with normal human genetic data,there are nine identical differential sites in DNA detection of diabetic retinopathy patients.We believe that diabetic retinopathy is related to diabetic retinopathy and need further verification.Traditional Chinese medicine has been used to treat diabetes for thousands of years.In traditional Chinese medicine,diabetes mellitus and diabetes mellitus are the closest,and they are often equated in modern medicine.DR belongs to the disease of diabetes mellitus.Most doctors call DR "diabetes mellitus".DR belongs to the category of "passerine","cataract","blindness","dim vision" and "clouds moving eyes".Deficiency of liver and kidney and deficiency of heart and spleen lead to loss of eyesight and orifice.After a lot of clinical practice,it has been proved that traditional Chinese medicine has a significant effect in the prevention and treatment of diabetic retinopathy,but at the same time,it is faced with the problem that the theoretical research progress of traditional Chinese medicine prescription is relatively slow because of its complex components and unclear mechanism,and it is difficult to achieve the preset goal by using traditional pharmacological experimental methods.With the gradual improvement of system biology and the vigorous development of network pharmacology combined with computer technology,there are interdisciplinary research methods,which have been gradually applied in the field of traditional Chinese medicine.The emergence of these new science and technology provides new methods and strategies for exploring the theory,clinical research and drug development of diabetic retinopathy.The research mode of network pharmacology has also chaned from "single drug,single target" to "disease-gene-target-drug".It is helpful to analyze,simulate and predict the pharmacological action mechanism of drugs,test and evaluate the clinical therapeutic effect and toxic and side effects of drugs by analyzing,simulating and predicting the biological system network as a whole.Systematic research and development of new Chinese medicine with multi-molecule and multi-target.Objects1 The high-resolution analytical method of mass spectrometry was used to verify 9 different sites of diabetic retinopathy in the previous study,to explore the specific high-expression gene locus in DNA of DR patients,and to analyze its biological mechanism.2 Using network pharmacology combined with bioinformatics,the biological mechanism of the treatment of diabetic retinopatthy with Prof.Xiaolin's experience party(sugar network)was analyzed to support the biological mechanism of highly expressed genes.Methods1 The whole blood samples of 40 patients with diabetic retinopathy were collected randomly in outpatient clinic and the quality of the samples was tested.After quality testing,nine SNP differential sites screened by previous studies were used as entry points.MassARRAY nucleic acid mass spectrometry was used to validate the specific gene loci with high expression.Six databases,NCBI database and ENSEMBL database were used to validate the specific gene loci.POLYS EARCH2 database;lncRNA SNP database(HGNC database queries the genes of the differential loci,through KEGG(Kyoto Encyclopedia of Genes and Genomes)database to find and analyze the related metabolic pathways(P<0.01).2 TCMSP,TCM Database and TCMID were used to find the chemical constitue nts of traditional Chinese medicines in sugar web recipe and the corresponding targets of diabetic retinopathy in OMIM database.Cytoscape 3.2.1 was used to construct the active ingredient-target interaction network of sugar-net recipe;OMIM and GAD database were used to find the gene data of diabetic retinopathy and related targets;Cytoscape 3.2.1 was used to construct drug-disease interaction network;STRING was used to construct PPI network between drugs and diseases,and screen for diabetic retinopathy.The selected targets were analyzed for GO enrichment,and metabolic pathways were screened in KEGG database to analyze the mechanism of Tangwang-fang in treating diabetic retinopathy.Results1 Forty patients with diabetic retinopathy were randomly enrolled in the study.The quality of blood samples collected was 39A and 1B.The concentration,purity and integrity of the samples met the experimental requirements.There were different-ces in the number of experiments.The study randomized 40 patients with diabetic reti-nopathy and performed MassARRAY nucleic acid mass spectrometry to verify 9 loci,namely rs9345283.rs738322.rs2844795,rs1727638,rs16984239,rs12505641.rs12142968.rs11243897,rsl 1175883.and the detection rate was verified.100%.through the database query rs738322 locus in the PLA2G6 gene.CDK-6 gene,PARP1 gene and APEX1 gene;rs11243897 locus in the AK8 gene;rs2844795 locus in the TRI-M31 gene;rs16984239 locus in the KCNS3 gene and RDH14 gene;The rs1727638 locus is located in the LINC01626 gene,the rs12505641 locus is located in the LOC105374524 gene,the rs9345283 locus,the rs12142968 locus,and the rs11175883 locus are not found.Analysis of metabolic pathways by KEGG database revealed that PLA2G6 is involved in TRP channel inflammatory mediator regulation,glycerol-phospholipid metabolism,ether lipid metabolism,arachidonic acid metabolism,alpha-linolenic acid metabolism,linoleic acid metabolism,vascular smooth muscle contra-ction,RAS signaling pathway.FcyR-mediated phagocytosis is involved in inflame-matory mediator regulation,lipid metabolism,contraction of vascular smooth muscle,regulation of autophagy,and phagocytosis;PARP1 is involved in NF-kB signaling pathway,programmed cell death,apoptosis,autophagy Sexual programmed cell death,Paraptosis,cell mitosis,cell swelling,and inflammatory response,immune response;APEX1 involved in base excision repair,and cell proliferation,differentiation and apoptosis and other cellular responses.2 TCMSP.TCM Database and TCMID were used to find the chemical consti-tuents of traditional Chinese medicines in sugar net prescription and the corres-ponding targets of diabetic retinopathy in OMIM database.Cytoscape software was used to construct the effective component-target interaction network of sugar web recipe;OMIM and GAD database were used to find gene data of diabetic retinopathy and related targets;Cytoscape software was used to construct drug-disease interaction network;STRING software was used to construct PPI network between drugs and diseases to screen out targets.GO enrichment analysis and metabolic pathway screening were carried out in KEGG database to analyze the mechanism of Tangwenfang in the treatment of diabetic retinopathy.3 By comparing the metabolic pathways screened in the first and second studies of comparative study,four common pathways were found:NF-kappa B signaling pathway.RAS signaling pathway,Fc gamma R-mediated phagocytosis and apoptosis.Conclusion1 Consider the PLA2G6 gene involved in the development of diabetic retinopathy through inflammatory mediator regulation,lipid metabolism,contraction of vascular smooth muscle,and regulation of cellular responses;PARP1 gene parti-cipates in the development of diabetic retinopathy through cellular responses,inflam-matory responses and immune responses;APEX1 participates through cellular respo-nses diabetic retinopathy develops.2 Through the analysis of network Pharmacology Method of intervention of Tangwangfang on diabetic retinopathy,it is speculated that Tangwenfang may play a multi-target intervention effect by inhibiting the activation of JNK signaling pathway in nervous system,inhibiting the inflammatory response induced by TNF-alpha,regulating the level of vascular endothelial factors,inhibiting cell apoptosis,improving microcirculation.Using network pharmacology to analyze the reliability of the mechanism of action of TCM compound is related to the data perfection of each drug and gene database,and the results of this study need to be further verified by clinical and animal experiments.