Data Mining Of Kidney Related Edema In Ancient Literature And The Experiments Of Pori A Cocos Related Formula And Main Component In Diabetic Kidney Disease

Object iveChronic kidney disease(CKD)has become one of the public health problems worldwide and raised global concerns.However,the etiology and pathogenesis of the progression of chronic kidney disease still have not fully understood.What' s more,due to the rapid increasing of risk factors,such as diabetes and hypertension,the incidence of CKD is growing and becoming a global public health problem.At present,the main clinical treatment of CKD were releasing symptoms,reducing complications,and dialysis replacement therapy.However,CKD not only cost a high burden of social economic but also bring a high burden for patients due to the long-term treatment period.Therefore,how to choose an effective way to delay the progression of CKD shows very important social and economic significance.In the process of inheritance and development of traditional Chinese medicine for thousands of years,the ancient literature of traditional Chinese medicine have recorded a large number of herbal,formula,acupuncture,cognition and treatment methods related to chronic kidney disease.In this wide spectrum area,the formula is the direct treatment that after physician' s clinical diagnosis and further inference in practice.Under the guidance of the theory of Chinese medicine,physicians could systematic arrange the herbals to make the herbals work through the most effective way.And make herbal medicine strengthen human body' s immune system and imbalance human body' s inner circumstances.Among the records in ancient literature,physicians have recorded a large number of effective herbals and formulas.They wrote down their clinical experience via ancient literature,which offer the original source for afterwards to learn.However,the concept of "kidney" in traditional Chinese medicine is not the same as the concept of kidney in modern medicine.The expression of kidney disease in ancient books often express as symptoms shows,such as edema,fatigue,proteinuria and so on.This cause some burdens for afterwards to learn and practice in clinical treatment.In this research,we choose the commonest symptoms of kidney related disease-edema to delving deep in its concepts,related knowledge,herbal medicines and formulas in ancient literature to study the unique experiments of treating kidney related edema in ancient literature.Moreover,we choose a classical formula,"Futu formula to interfere the diabetic kidney disease rats to study the effectiveness of this formula and using RNA-sequencing to explore the potential mechanism.And we choose Pachymaran,the main component of Porio cocos,to interfere the high glucose induced endothelial cell dysfunction to prove the effectiveness and mechanism.MethodsThis study included two parts.For the ancient literature mining,we choose a technique in Artificial Intelligence-"knowledge graph",a constructed based on the basis of computer ontology.We combine knowledge graph with human power to identify and excavate the kidney related edema.The key word is "shuizhong or fuzhong".We search in the ancient literature database which includes for more than 1100 ancient literature.After first step searching,a total of 11893 articles were found,we screened and sorted one by one to set the exclusion criteria shelter and screen out the non-renal related edema.The descriptions which could not be excluded at first were keep included and distinguish in next round screening.5257 records of obvious non-renal edema were screened out,and 6636 valid articles were included.6636 valid articles were included for screening to further exclude"local edema","hepatic edema","nutritive edema" and cardiac edema".For non-renal edema related records,they were classified into"prescription","pathogenic pathogenesis","symptoms","management" and"herbal medicine" according to the content of the records.Later on,we settle the requirement of the property ontology in the unified knowledge extraction format such as " source "," efficiency attendance",symptoms","drug composition"," manufacturing method",etc.Extracting useful information from the articles of classification marks.And later we classified the articles,standardized drug names,method and usage according to the knowledge map model.Moreover,the knowledge element concept is revised and semantically labeled according to the ontology analysis.The construction of the ontology knowledge base and the excavation of ancient edema of renal edema was carried out to explore the rules of ancient literature on renal diseases.Two prescriptions commonly used in clinical practice,Six-ingredient Rehmannia Pill and Futu dan were used to explore the classification of the prescription layer,the drug pair(Fuling and shanyao),and the single herbal,Fuling,were analysis in next step experimental study.The second part is the basic experimental study.Due to the high frequency and importance of Porio cocos in the renal related edema prescription,the experimental study selected the diabetic kidney disease and rats and high glucose damaged cell experiments to study the experimental utility of Porio cocos as the main prescription.The experiment is divided into two parts:animal experiment and cell experiment.In the animal experiment,the typical steroidal remedy obtained from the renal edema knowledge base was selected,and the most common primary disease of the kidney disease was selected for related experimental research.A rat model of diabetic nephropathy was established by streptozotocin unilateral nephrectomy.and was treated with Futu formula for 10 weeks,the research data included blood analysis,urine analysis and RNA-seq data in order to analysis the therapeutic effect and possible mechanism of Futu formula on diabetic nephropathy rats.In the cell experiment,the main component of Poria cocos,Pacymaran,was used to treat the high glucose-induced endothelial cell model,and the related indicators were measured by MTT and flow cytometry to further verify the experimental results.ResultsThe results of data mining in kidney related edema ancient literature showed that the prescriptions included 4013 formula,1052 herbals,and the top 10 traditional Chinese medicines used in the treatment of kidney related edema were Tonifying Medicinal?Heat-clearing drugs Medicinal?Blood-activating and Stasis-resolving Medicinal?Antitussive and Antiasthmatic Medicinal?Hemodtstic Medicinal?Wind-damp-dispelling Medicinal?Qi-regulating Medicinal?Damp-draining Diuretic?Exterior-releasing Medicinal?Interior-warming Medicinal.The most frequently used Chinese medicine subcategories(compared with subdivisions in large categories,such as Tonifying Medicinal are divided into Tonifying Qi Medicinal,Tonifying Yang Medicinal,Tonifying Yin Medicinal,etc.),the top five are:Blood-activating and Stasis-resolving Medicinal?Wind-damp-dispelling Medicinal?Tonifying Yang Medicinal?Qi-regulating Medicinal?Tonifying Qi Medicinal.In the frequency statistics of traditional Chinese medicine,the top ten herbal drugs used are Poria cocos,Pericarpium citri reticulatae,Rhizoma alismatis,Rhizoma atractylodis macrocephalae,Radix glycyrrhizae,Semen descurainiae,Radix aucklandiae,Cortex mori,Semen arecae,Radix stephaniae tetrandrae.Among them,Poria cocos(Fuling)is the only traditional Chinese medicine that uses more than one thousand frequencies.The results of excavation of the classical prescription Six-ingredient Rehmannia Pill included(higher or equal to 0.5)9 pairs:"Fuling-Shanyao(0.52)" and "Fuling-Zexie(0.59),"Fuling-Mu Dan Pi(0.57)","Fuling-Shu Di Huang(0.5)","Fuling-Shan Zhu Yu(0.52)","Shu Di Huang-Mu Dan Pi(0.52)","Shu Di Huang-Ze Xie(0.64)","Ze Xie-Mu Dan Pi(0.55)" and "Ze Xie-Shan Yao"(0.5).Among them,Fuling is associated with all other 5 drugs ? 0.5,indicated Poria cocos(Fuling)is an important herbal.In the case of clinical kidney related edema,Six-ingredient Rehmannia Pill are mainly used in modern medical definition of male diseases(seminaria,spermatorrhea,impotence,leakage,ejaculation)and common symptoms of in nephrology,such as hematuria,proteinuria,waist and knee pain,frequency of urination and so on.Analysis of related drugs,symptoms and pathogenesis of Six-ingredient Rehmannia Pill by knowledge graph.It was found that in addition to the well-known functions such as remove dampness and promote urination,strengthening the spleen and calm the heart.In kidney related edema disease,Poria cocos is also used for hematuria or proteinuria,leakage,language sputum,qi deficiency and spontaneous sweating,rheumatism and other symptoms.This broadens the clinical re-recognition of Poria cocos and realizes the function from "the use of clinical prescriptions-literature data mining-clinical prescriptions".On the basis of the single-flavored drug,the machine-based learning and manual proofreading were carried out to realize the deduction of the core drug pair of Fuling-Shanyao-Wuweizi" and the recommendation "Futu Dan" prescription.Excavation of the symptoms related to Futu Dan formula can be found that the top ten symptoms are "nocturnal emission","diarrhea""spermatorrhoea"," ematuria or proteinuria"," vomiting","impotence",“waist and knee pain”,"frequent urination","thirst" and poor appetite".Due to there is no specific clinical disease associated with kidney related edema in ancient literature,we analysis with clinical epidemiological data and found that diabetic nephropathy is the top 1 reason for chronic kidney disease,establish an animal model,experimental excavation of the most important cause of chronic kidney disease,therefore we established a diabetic nephropathy rat model by unilateral nephrectomy plus streptozotocin.And later the rats was treated with Futu Dan formula for 10 weeks.After that;blood analysis,urine analysis and RNA-seq were used to analysis the therapeutic effect and possible mechanism of Futu Dan formula on diabetic nephropathy rats.Through the blood and urine analysis indicators,it was found that Futu Dan formula can effectively reduce serum creatinine in diabetic nephropathy rats.Gene analysis of normal group rats and diabetic nephropathy rats were analysis by RNA-seq to screening differential genes,refining the influencing pathways and related genes of diabetic nephropathy rats,and found that G0:0001872 protein is(1-3-?-D-polysaccharide binding protein)could be the Diabetic nephropathy related affecting protein.Further analysis in the pathway for further influences found that the pathways include aging pathway,Glycan biosynthesis and metabolism,amino acid metabolism,tumor,carbohydrate metabolism and other pathways,and can affect multiple system functions(cardiovascular system,digestive system,endocrine system,etc.)It affects cell growth,exercise,and death pathways,and has related effects on immune diseases and infectious diseases.Cell experiments:The results of RNA-seq indicated that(1-3-?-D polysaccharide)could be a influence factor.Therefore we choose the main component of Poria cocos,Pachymaran(1-3-?-D polysaccharide)to intervene in high glucose-induced endothelial cell injury model in order to explore the influence of Klotho protein.By MTT analysis and flow cytometry to establish the model to determine the high glucose intervention concentration,high glucose intervention time,cell starvation concentration,and pachymaran intervention concentration.Later on we use flow cytometry found that high glucose can lead to inhibition of endothelial cell Klotho protein expression and injured endothelial cell function.Pachymaran can protect endothelial cells from aging pathways by promoting the expression of Klotho protein,and delaying the damage of high glucose endothelial dysfunction.ConclusionIn this study,we first establish a kidney related formula database by using ontology and semantic network,and study a complete study from literature mining,animal experiments and cell experiments.From the analysis of the ancient literature formula,to classical formula("Six-ingredient Rehmannia Pill" and "Fu Tu Dane),and to Poria cocos,the main component of Poria cocos,we have try to explored the "formula-herbal drug-main ingredients" pattern.The most frequently used in kidney related edema is Poria cocos,later on we broadened the cognition of the ancient recipes related to the typical prescriptions of Six-ingredient Rehmannia Pill and Fu Tu Dan.From the perspective of animal experiments,it was found that Fu Tu Dan oral administration can improve the corresponding renal function indexes in rats with diabetic nephropathy.Pachymaran could protect against high glucose endothelial damage from aging pathways.