Research On The Regularity Of Antipyretic Drugs In The Treatment Of Type 2 Diabetes And Its Mechanism Of Regulating Intestinal Flora

Objective1.Based on the data mining of prescriptions related to the treatment of diabetes in ancient Chinese medical books and modern literature,this paper studies the rules and characteristics of the prescriptions and heat-clearing drugs used by ancient and modern doctors in the treatme nt of diabetes.In addition,through the comparative study of Classification of Medical Formul as(Yi Fang Lei Ju)and Chinese Medical Code(Zhong Hua Yi Dian),the clinical value of Cla ssification of Medical Formulas as a large-scale book of formulas was revealed.2.Through the data mining of modern clinical medical records,type and compatibility rul es of commonly used heat-clearing traditional Chinese medicines(TCM)for the treatment of t reatment of type 2 diabetes mellitus(T2DM)by Professor ??? were revealed.Such data pr ovides ideas and reference for clinical application of heat-clearing Chinese medicine in the tre atment of T2DM.3.Based on the theory of intestinal flora,the microbiological targets underlying the hypo glycemic effect of heat-clearing drugs could be explored from the level of biotaxonomic "gen us" via Illumina Miseq PE300 sequencing platform and technology,which was utilized in ord er to reveal the mechanism of action for typical heat-clearing drug Coptis chinensis' hypoglyc emic effect.Methods1.Literature research.Ancient literature research involved extracting and organizing all"wasting-thirst(xiaoke)" treatment prescriptions from the entirety of Classification of Medica l Formulas' xiaokemen chapter and Chinese Medical Code's relevant sections.First,frequenc y analysis was used to determine the most commonly recurring qi,flavor,meridians,formulati ons,and drugs;all prescriptions were screened for heat-clearing effect as inclusion criteria,an d the individual heat-clearing drugs were all counted.Association rule and cluster analysis we re used to study the compatibility rule of heat-clearing drugs for treating wasting-thirst.For m odern literature research,current literature related to treatment of T2DM via TCM were retriv ed from the CNKI,Wanfang and Weipu databases;studies were selected based on specific inc lusion and exclusion criteria.Frequency analysis was utilized to obtain corresponding statistic s regarding four qi,five flavor,meridians,formulations,and drugs;heat-clearing drugs were c ounted from applicable prescriptions as according to inclusion criteria based on heat-clearing effect.Association rules and cluster analysis were performed to study compatibility rules for t he use of heat-clearing drugs in T2DM treatment by modern physicians;in addition,most co mmonly used dosages for the most frequently used heat-clearing drugs were assessed.2.Clinical research.396 medical records of Professor ??? for the diagnosis and effect ive treatment of T2DM were collected.First,the terminology for patient symptoms,tongue,p ulse,and prescription drug names were standardized as a medical records and treatment decoc tions database was created.Afterwards,statistical analysis was conducted regarding patient co nditions,prescription drugs,four qi,five flavors,and attributed meridians.Using the TCM inh eritance assistant platform(version 2.5)developed by the Chinese Academy of Chinese Medic al Sciences,association rules,complex system entropy clustering analysis,and unsupervised e ntropy hierarchical clustering analysis were conducted on medical data fulfilling the heat-clea rising inclusion criteria for drugs in order to determine Professor ??? prescription compati bility rules for T2DM treatment with heat-clearing drugs.Further statistical analysis was cond ucted to determine medical characteristics and dosages for drugs used to treat different heat sy ndromes3.Experimental research.18 spontaneous gene-deficient T2DM model mice(db/db mice)were randomly divided into model,Coptis chinensis,and metformin groups;6 littermate non-diabetic mice(db/m mice)were formed the normal(control)group.Mice from the Coptis chin ensis and metformin groups were given aqueous solutions of Coptis chinensis and metformin respectively;pure water was given to mice of the model and normal groups;intervention time was 8 weeks.Changes in blood glucose and body weight of subjects were observed during the experiment.Fecal samples were collected at week 8 prior to termination;Illumina Miseq PE300 sequencing platform was then used to analyze DNA sequencing and clustering of sample c olonies in region 16S rDNA V3-V4,utilizing the silva132/16s_bacteria species classification database.Composition,abundance,and diversity of intestinal flora was therefore analyzed in-depth at the biotaxonomic "genus" level.Results1.Literature research results indicated the properties of drugs ancient physicians used to t reat wasting-thirst were mainly cold,sweet,and bitter,corresponding to primarily the lung an d stomach meridians.Drug formulations were primarily decoctions,pills,and powders,The m ost commonly used heat-clearing drugs were trichosanthin,Ophiopogon japonicus,Coptis chi nensis,Anemarrhena rhizome,Rehmannia root,licorice,gypsum,Scutellaria baicalensis,and Rehmannia bark.Association rules results with the highest support and confidence were Scute llaria baicalensis,Ophiopogon japonicus,Radix rehmanniae,Radix ophiopogonis,Anemarrhe nae Rhizoma Coptidis,Radix Ophiopogonis,Radix Astragali,and ginseng.The core drug co mbination comprised of trichosanthin,Coptis chinensis,Ophiopogon japonicas,and ginseng.Cluster analysis revealed a total of 6 plausible new prescription combinations,including trich osanthin,Ophiopogon japonicus,anemarrhen,baicalein,mulberry bark,Rehmannia bark,Pue raria root,cohosh,Poria god,red tuckahoe cocos,ginger,bamboo leaf,stir-fried licorice,Copt is chinensis,trichosanthin,king gourd root,oyster,Sophora flavescens,endothelium corneum amylase,and red lead.Current literature research results showed that the medicinal properties of TCM drugs prescribed by modern physicians to treat T2DM were mainly chilly,sweet,and bitter,attributed to primarily the spleen and lung meridians.Prescriptions were mainly formul ated as decoctions and capsules.Most commonly prescribed heat-clearing drugs were Rehma nnia root,Coptis chinensis,trichosanthin,Ophiopogon japonicus,Radix scrophulariae,anema rrhenae,licorice,and red peony root.Association analysis results revealed that the rules with t he highest support and confidence were Radix Rehmanniae,Radix Astragali,Radix Salviae M iltiorrhizae,Radix Astragali,Radix Rehmanniae,Radix Salviae Miltiorrhizae,and Radix Astr agali.The modern core drug combination was trichosanthin,Coptis chinensis,Ophiopogon ja ponicus,Rehmannia root,Astragalus membranaceus,yam,pueraria root,and Salvia miltiorrhi za.Cluster analysis revealed 5 new potential prescriptions,which included gardenia,Scutellar ia baicalensis,astragalus,Radix scrophulariae,Salvia miltiorrhiza,Radix scrophulariae,Atrac tylodes macrocephala,Poria cocos,dogwood,Alisma orientalis,and peony bark.2.In the clinical research section,396 prescriptions from the 396 modern medical records were included.Most frequently occurring symptoms were,from most common to least,fatigu e,dry mouth,insomnia,bitter taste,and mild lumbago.Stagnant-heat,yin deficiency,and qi d eficiency syndromes were the most prevalent,followed by blood stasis and stagnant-heat synd rome;the heat syndromes and most related syndromes(stagnant-heat,yin deficiency,accumul ated heat,phlegm-heat,damp heat,liver-yang,and heat-toxicity syndromes)accounted for 50.66%of the total diagnosed syndromes.For prescription drugs,the main medicinal properties were cold,bitter,and sweet,primarily attributed to the liver and spleen meridians.The most fr equently prescribed heat-clearing drugs were Scutellaria baicalensis,Coptis chinensis,Rehma nnia glutinosa,Licorice,Prunella vulgaris,Radix Paeoniae Rubra,Radix rehmanniae,Fructus burdock,and Rhizoma Anemarrhenae.Association rules results with the highest support and c onfidence included Pueraria lobata,Salvia miltiorrhiza,Radix Rehmanniae,Radix Scutellaria e,Cortex Lichi,Cortex Rehmanniae,Radix Salviae Miltiorrhizae,Herba Anemarrhizae,Corte x Litchi,Radix Puerariae,Radix Salviae Miltiorrhizae,Radix Bupleuri,Scutellaria baicalensis,and Pericarpium Citri.The core drug combination was Pueraria lobata,Salvia miltiorrhiza,B upleurum chinensis,Scutellaria baicalensis,Paeonia lactiflora,Coptidis chinensis,Rehmannia cortex,Euonymus alatus,Litchi seed,and Crassorhizoma agrimonis.Complex entropy cluste ring analysis revealed 8 groups of potential core drug combinations,including Ophiopogon ja ponicus,Schisandra chinensis,Radix et Rhizoma et Rhizoma Paeoniae Alba,Radix Paeoniae Rubra,Licorice,Cortex Reticulatae,Herba Crassiae,Litchi seed,and silkworm droppings.Re sults of unsupervised entropy hierarchical clustering analysis revealed four new potentially co mpatible prescription combinations,including Ophiopogon japonicus,Plumula nelumbinis,lo tus seed,Schisandra chinensis,Pseudostellaria heterophylla,Paeonia lactiflora,Radix Paeonia e rubra,licorice,Panax notoginseng,Viola vulgaris,and Buddleja officinalis.Via statistics of t he medication of heat and related syndromes,the most commonly used heat-clearing drugs for each syndrome were obtained:Scutellaria baicalensis,Prunella vulgaris,and Radix Paeoniae rubra were most frequently used for treating stagnant-heat syndrome;wolfberry bark,Rehman nia root,Anemarrhenae rhizoma,and Radix Scutellariae were primarily prescribed for yin def iciency;Scutellaria baicalensis,Coptis chinensis,honeysuckle,forsythia,and dandelion were the most frequently used drugs for heat-toxicity syndrome3.Results of animal experiments showed that the blood glucose index of spontaneous gen e-deficient T2DM mice given Coptis chinensis(Coptis chinensis group)significantly decrease d compared with that of the model(pure water)and metformin groups(P=0.002,P=0.033).W eight was measured for mice before and after experimentation.The weight of mice in the mod el,Coptis chinensis,and metformin groups were significantly different from that of non-diabe tic mice in the normal group(P value=0.000),but there was no significant difference among the three groups of diabetic mice(P value>0.05).Based on 16S rDNA sequencing results,in comparison with the model group,mice in the Coptis chinensis group exhibited a 1.)decrease d abundance and diversity of intestinal flora,2.)increased abundance of Bacteroidetes,Proteo bacteria,Cyanobacteria,and Verrucomicrobia bacteria,3.)decreased abundance of Firmicutes,Actinobacteria,Patescibacteria,Epsilonbacteraeota,Tenericutes,and Deferribacteres bacteria,and 4.)increased Bacteroidetes/Firmicutes ratio;at the biotaxonomic "family" and "genus" le vel,5.)increased abundance of Akkermanisia and Akkermansiaceae bacteria,while 6.)Alistip es,Blautia,Roseburia,Helicobacter,Papillibacter and Ruminococcaceae bacteria abundance decreased.Correlation analysis of intestinal flora and its effect on blood glucose indicated that Akkermansia was negatively correlated with blood glucose,with statistical significance(P=0.00026;Q=0.00866).Conclusion1.Literature studies showed that historically,heat-clearing drugs were used by physicians to treat wasting-thirst,usually in conjunction with qi and yin tonifying drugs.The most com monly used heat-clearing drugs were trichosanthin,Coptis chinensis,and Anemarrhenae Rhiz oma.Modern physicians likewise emphasize the usage of heat-clearing drugs in T2DM treatm ent,but in contrast,normally combine it with blood circulating and stasis-removing drugs.Th e most frequently prescribed heat-clearing herbs by modern physicians were Radix Rehmanni ae,Coptis chinensis and,trichosanthin.2.Analysis of modern clinical cases revealed that the most common heat syndromes of T 2DM were stagnant-heat,yin deficiency,and heat-accumulation.Professor??? most frequ ently prescribed the heat-clearing Scutellaria baicalensis,Coptis chinensis,Digupi,and Prunel la vulgaris drugs for T2DM treatment.Such TCM heat-clearing drugs were often used in com bination with drugs to invigorate the spleen,tonify qi and nourish yin,circulate blood for stas is removal,relieve liver qi stagnation,dry dampness and resolve phlegm.3.Experiment results showed that the hypoglycemic mechanism of Coptis chinensis,as a representative of TCM heat-clearing drugs,is related to the increased abundance of Akkerman sia bacteria,alongside the reduced abundance in Alisipes,Blautia,Helicobacter,Papillibacter,Roseburia,and Ruminococcaceae bacteria,thereby resulting in improved structural compositi on and lowered diversity of intestinal flora.However,considering the multi-target mechanism of TCM,further research into the mechanism of heat-clearing TCM drugs for T2DM treatme nt is warranted for further clarification.4.Currently,research level of traditional medicine in Thailand is relatively weak.Increasi ng awareness and publication of research results for the prevention and treatment of T2DM us ing TCM would be conductive in improving the quality of TCM clinical diagnosis and treatm ent of T2DM within Thailand.