Analysis Of Professor Tong Xiaolin’s Medication Rule In The Treatment Of Glycolipid Metabolism Disorder And Exploration Of The Mechanism Of Core Drugs

The incidence rate of diseases related to the disorder of glucose and lipid metabolism,such as obesity,diabetes,and dyslipidemia,is on the rise year by year,which aggravates the progress of vascular complications,seriously endangers human health,and increases the burden of medication for glycolipid comorbidities.It is urgent to find more efficient,safe,and comprehensive treatment plans and drugs that can regulate glucose and lipid metabolism disorders.Professor Tong Xiaolin is deeply involved in the field of traditional Chinese medicine in the treatment of metabolic diseases,and has accumulated a lot of experience in prescription medication,but lacks systematic summary and induction.Therefore,this study summarizes Professor Tong Xiaolin’s medication rules for the treatment of glucose and lipid metabolism disorders through the method of data mining,refines the core prescription of glucose reduction and lipid regulation,and further explores the mechanism of its regulation of glucose and lipid metabolism disorders using modern omics technology.Objective1.Through data mining of Professor Tong Xiaolin’s medical record on the treatment of glycolipid metabolism disorder,summarize and summarize the medication rules and formulation ideas of different courses of disease,and refine the core prescription,with a view to providing a new treatment plan for clinical treatment of glycolipid metabolism disorder.2.By establishing a mouse model of glucose and lipid metabolism disorder,and using modern omics research methods,further explore the mechanism of action of the core prescription of JTTZ.Methods1.Analysis of Professor Tong Xiaolin’s medication rules in the treatment of glucose and lipid metabolism disorders based on data mining of outpatient medical records: based on the criteria for admission and discharge,the medical record data of obese type 2 diabetes patients with dyslipidemia who visited Professor Tong Xiaolin’s outpatient clinic from January 2011 to December 2021 were collected,and a database was established.The medical record data were entered uniformly after standardization,and frequency statistics were conducted using Microsoft office Excel 2007,SPSS Modeler 18.0 and SPSS Statistics22.0 Association rule analysis and cluster analysis,summarize and summarize Professor Tong Xiaolin’s medication rules for the treatment of glucose and lipid metabolism disorders,and refine the core prescription.2.Exploration of the mechanism of the core prescription of JTTZ to regulate glucose and lipid metabolism disorder based on the omics technology: to establish a mouse model of glucose and lipid metabolism disorder induced by high-fat diet,to detect the relevant indicators of glucose and lipid metabolism,and to verify the regulatory effect of the core prescription of lowering blood sugar and regulating blood lipid on the disorder of glucose and lipid metabolism;LC-MS targeted metabonomics technology was used to detect the changes of metabolic products in mouse adipose tissue;RNA-seq transcriptome technology was used to detect the changes of transcriptional spectrum;Combined with targeted metabolomics and transcriptomics,we obtained the key genes and metabolites of the core prescription of JTTZ and regulating blood lipid metabolism,and verified the key genes by RT-PCR to reveal its regulatory mechanism.Results1.Analysis of Professor Tong Xiaolin’s medication rules in the treatment of glucose and lipid metabolism disorders based on data mining of outpatient medical records:(1)High-frequency drugs: the high-frequency drugs with a disease course of less than 5 years are Mori Follum,Coptidis Rhizoma,Hedysarum Multijugum Maxim,Anemarrhenae Rhizoma and Citrus Reticulata;The course of disease is 5-10 years for Hedysarum Multijugum Maxim,Zingiber Officinale Roscoe,Anemarrhenae Rhizoma,Mori Follum and Hirudo Nipponica;TThe drugs with a course of disease of 10-20 years or more are Hedysarum Multijugum Maxim,Hirudo Nipponica,Coptidis Rhizoma,Anemarrhenae Rhizoma and Zingiber Officinale Roscoe.(2)Association rule analysis: The two commonly used drug combinations with disease duration less than 5 years are mainly Anemarrhenae Rhizoma and Radix Paeoniae Rubra,Curcumae Rhizoma and Radix Notoginseng,Mori Follum and Mori Cortex;The combination of three drugs is Mori Follum,Citrus Reticulata and Anemarrhenae Rhizoma;Mori Follum,Anemarrhenae Rhizoma,Hedysarum Multijugum Maxim;Fritillariae Thunbrgii Bulbus,Curcumae Rhizoma,Radix Notoginseng;Coptidis Rhizoma,Anemarrhenae Rhizoma,Mori Follum and Hedysarum Multijugum Maxim are the core networks;The course of disease is 5-10 years.The combination of two drugs is Hedysarum Multijugum Maxim and Hirudo Nipponica;Hedysarum Multijugum Maxim and Atractylodes Macrocephala Koidz;Hedysarum Multijugum Maxim and Epimrdii Herba;The commonly used combination of three medicines is Zingiber Officinale Roscoe,Radix Paeoniae Rubra and Anemarrhenae Rhizoma;Anemarrhenae Rhizoma,Mori Follum and Zingiber Officinale Roscoe;It has formed a key drug network with Hedysarum Multijugum Maxim as the core and Epimrdii Herba,Radix Rhei Et Rhizome and Hirudo Nipponica;The combination of two drugs commonly used for 10-20 years or more is mainly Hedysarum Multijugum Maxim and Hirudo Nipponica;The combination of three drugs is Radix Salviae,Radix Rhei Et Rhizome and Hirudo Nipponica.(3)Cluster analysis: The drug cluster drug combination at different stages of the disease contains the meaning of Xiaoxianxiong decoction-Tongshi Huangchi decoction-Qixue decoction + Tongshi Qidan Junzhi decoction.(4)Exploration of core prescription: based on the theoretical basis of "clearing heat and removing ointment and reducing turbidity" to treat the disorder of glucose and lipid metabolism,through data cluster analysis of 1021 prescriptions collected,a combination of drugs with Coptidis Rhizoma,Anemarrhenae Rhizoma and Monascus Purpureus Went as the core has been formed,and the core prescription of JTTZ has been refined after consulting the tutor.2.Exploration of the mechanism of the core prescription of JTTZ to regulate glucose and lipid metabolism disorder based on the omics technology:(1)The regulatory effect of JTTZ on the disorder of glucose and lipid metabolism: JTTZ can significantly reduce the body weight,fasting blood glucose level and serum TG level of mice with disorder of glucose and lipid metabolism(P<0.0001);(2)Targeted metabolomics results: By setting the screening criteria for different metabolites,there are 17 significantly different metabolites between the model group and the control group,of which 5 are up-regulated and 12 are down-regulated,indicating that after the induction of high-fat diet,the metabolism of fatty acids,carbohydrates,short-chain fatty acids and bile acid metabolites in mice is disordered.There are 15 metabolites with significant difference between JTTZ and model group,of which 5 are up-regulated and 10 are down-regulated,indicating that after JTTZ intervention,the metabolites of short-chain fatty acids,fatty acids,bile acids and amino acids in mice have significantly changed.Among them,fatty acid and bile acid metabolites have the most significant changes.Further,based on the analysis of enrichment pathway of differential metabolite KEGG,the main metabolic pathway is the biosynthesis pathway of unsaturated fatty acid;(3)Transcriptomics results: By setting the screening criteria for differentially expressed genes,4553 differentially expressed genes were screened from the control group and the model,of which 2148 were up-regulated and 2405 were down-regulated;The JTTZ group and the model group were significantly different in 434 genes,334 genes were up-regulated and 100 genes were down-regulated.Further interaction analysis of 265 common differential genes found 7 genes closely related to JTTZ intervention in glycolipid metabolism disorder.GO enrichment analysis and KEGG enrichment pathway analysis showed that the biological process of triglyceride metabolism,PPAR signal pathway,glyceride metabolism and cholesterol metabolism pathway had significantly changed.(4)Explore the mechanism of JTTZ based on the joint analysis of transcriptomics and metabonomics: by integrating and analyzing the data of transcriptomics and targeted metabonomics,construct the regulatory network between differential genes and differential metabolites,and obtain key metabolites such as 2-hydroperoic acid,CA,Palmitoleic acid,γ-Linolenic acid,Linoelaidic acid,α-Linolenic acid.And key genes of Slc2a4,Adipoq,Lipe,Pnpla3,Dgat1,Gsta3,Plin4 and Thrsp were further verified by RT-PCR.The results showed that Slc2a4,Plin4,Pnpla3,Dgat1 showed a significant upward trend after JTTZ intervention(P<0.05).Conclusion1.In the early stage of diabetes,the main drugs were clearing stagnation heat,such as Coptidis Rhizoma,Anemarrhenae Rhizoma and Mori Follum.In the stage of diabetes and complications,the main therapeutic method is to nourish qi,promote blood circulation,and dredge meridians,and commonly used Hedysarum Multijugum Maxim,Radix Salviae,Hirudo Nipponica,etc.Based on Professor Tong Xiaolin’s basic treatment methods for the treatment of glucose and lipid metabolism disorders,core formulas and medicines mainly composed of Coptidis Rhizoma,Anemarrhenae Rhizoma and Monascus Purpureus Went were obtained through data mining.Combined with the theoretical guidance of “clearing the stomach and purging heat,eliminating ointments,and reducing turbidity”,the core formulas and medicines for reducing glucose and regulating lipid were summarized.2.JTTZ can significantly reduce the body weight,blood glucose and blood lipid levels of model mice,and effectively regulate the glucose and lipid disorders of mice.It can significantly regulate the levels of fatty acids and bile acid metabolites,and the biosynthesis pathway of unsaturated fatty acids.Seven genes and multiple enrichment pathways closely related to JTTZ’s intervention in glucose and lipid metabolism disorders were found,among which the biological process of triglyceride metabolism,PPAR signal pathway and glyceride metabolism were the most significant.Through omics integration and validation analysis,It was confirmed that JTTZ could regulate the expression of Slc2a4,Plin4,Pnpla3,Dgat1 and regulate 2-hydroxycaproic acid,CA,Palmitoleic acid,γ-Linolenic acid,Linoelaidic acid,α-linolenic acid,thus play the role of regulating the disorder of glucose and lipid metabolism.