| Objective:1.Based on data mining technology,analyze Professor Liu Yanjing’s medication rules in the treatment of NAFLD,in order to provide reference for clinical practice.2.Carry out animal experiments to determine the therapeutic effect of Zhishi Daozhi Decoction on NAFLD model mice,and explore the mechanism of its treatment of this disease.Methods:1.Through the ancient and modern medical record cloud platform(V2.3.5),data mining and medication rules analysis of NAFLD cases treated by Professor Liu Yanjing were carried out.2.The mice were randomly divided into control group C(n=13,given normal diet)and high-fat model group(n=53,given high-fat diet).After 10 weeks,the blood biochemical and pathological tests showed that the model was successful.The high-fat group was randomly divided into model group M,Polyene phosphatidylcholine group P,Emodin E,Berberine B,and Zhishi Daozhi decoction group Z,and after 4 weeks of treatment by gavage,the body weight,serum liver function,blood lipids,and liver pathology were tested,The fecal intestinal flora was detected by 16 Sr RNA,the content of fecal short chain fatty acids was detected by gas chromatography,and the expression of TJ protein,occludin and ZO-1,was detected by immunofluorescence.Results:1.Clinical studyMale patients are slightly more than female patients;The symptoms were mainly hypochondriac discomfort,fatigue and gastric distension;Damp and heat are the most common syndromes;Red tongue,yellow and greasy fur and slippery pulse are the most common;Prescription high-frequency drugs include Fructus Aurantii,Atractylodes macrocephala,Bupleurum chinense,Alisma orientalis,Shenqu,Scutellaria baicalensis,Rhizoma coptidis,rhubarb,Poria cocos,and Aucklandia;In the prescription,cold drugs are slightly more than hot drugs,and bitter drugs account for the largest proportion;The meridians of drugs are mainly spleen,stomach,lung,heart,large intestine and gallbladder;Its efficacy is mainly to clear heat and dry dampness,promote water penetration and damp,purge fire and detoxify;Drug association analysis showed that there were more drug pairs centered on Fructus Aurantii;According to the drug cluster analysis,the most complete expression was Poria cocos,Bupleurum chinense,Shenqu,Atractylodes macrocephala,Alisma orientalis,Rhubarb,Fructus Aurantii,Scutellaria baicalensis,and Coptis chinensis;Through complex network analysis,it is concluded that Professor Liu Yanjing’s core traditional Chinese medicine prescription for treating NAFLD is Zhishi Daozhi Decoction plus or minus.2.Experimental study(1)Body weight,blood biochemistry,liver pathology(1)Body weight: After 10 weeks of high-fat modeling,there was a significant difference between group M and group C(P<0.01).After 4 weeks of treatment,the Z group was better than E group than B group and than P group.(2)Blood biochemistry: liver function: high-fat model significantly increased ALT and AST(P<0.01,group C vs group M).ALT and AST were significantly optimized in P group(P<0.01,group P vs group M).The optimization of ALT in groups B and Z were acceptable(P<0.05,group B and Z vs group M),but group E had no effect.Lipid: hyperlipidemia model significantly increased CHO and TG(P<0.01,group C vs group M).In terms of improving CHO index,groups B and Z were effective(P<0.05,group B vs group M),(P<0.01,group Z vs group M).(3)Liver pathology: Compared with group C,group M had obvious fat deposition,while other groups were between the two.(2)Intestinal flora(1)Intestinal flora diversityAlpha analysis(Chao1 and Shannon index)showed that there was no difference between groups M and C,while the level of group B was significantly lower than that of groups C,M and P(P<0.01).Beta analysis(PCo A and Amova statistics)showed that the sample aggregation in each group was good,and there was a difference with other groups.There was a difference in the composition of flora between groups C and M(P<0.05),but there was no difference between groups Z and C(P>0.05).There were differences between group B and other groups(P<0.05).(2)Community compositionPhylum level: Bacteroides,Bacteroides and Proteobacteria are the dominant phyla of each group of samples.Compared with group M,groups Z and C were at a higher level in Bacteroides.group B was at a very high level in Proteus.Genus level: Faecalibaculum,Bacteroides,Blautia,Colidextribacter,Clostridioides and Citrobacter are the dominant bacteria in each group.In terms of Faecalibaculum and Bacteroides,groups C and Z were at a high level,while the other groups were at a low level;As for Blautia and Colidextribacter,groups C and Z were at a low level,while the other groups were at a high level;For Clostridioides and Citrobacter,only group B was at a high level.(3)LEfSeThe difference between groups C and M was due to the up-regulation of Faecalibaculum,and the decline of Colidextribacter and Blautia;The reason for the difference between groups Z and M was similar to that of groups C and M;The difference between groups P and M was due to the up-regulation of Lactobacillaceae and the decrease of Bacteroidales;The difference between groups E and M was due to the up-regulation of Clostridioides and the decline of Lactobacillus;The reason for the difference between groups B and M was the up-regulation of Citrobacter and Clostridioides,and the decline of Lactobacillus,Blautia and Faecalibaculum.(3)Short chain fatty acidsAcetic acid,propionic acid and butyric acid dominate the composition of short chain fatty acids.In terms of the content of the three,the situation of groups Z and C was similar,and they were at a high level.However,the situation of groups P,E and B was similar to that of group M,which were at a lower level.(4)Ocludin,ZO-1 proteinIn the expression of two TJ proteins,groups Z and C maintained good morphological expression and high expression,and there were no statistical difference between the two groups(P>0.05).However,the situation of groups M,P,E and B was not good,and there were significant differences between the four groups and C group(P<0.01).Conclusion:1.Clinical studyProfessor Liu Yanjing believes that the disease is located in the liver,which is closely related to the spleen,stomach,gall bladder and large intestine.The pathogenesis of the disease is based on the accumulation of dampness and heat to make the stool abnormal.The treatment is mainly based on the traditional Chinese medicine of clearing heat and dampness,purging constipation and purging fire,and supplemented by the sweet and warm traditional Chinese medicine of invigorating the spleen and promoting water,digesting and dissolving accumulation.The core prescription is often the addition and subtraction of Zhishi Daozhi Decoction.2.Experimental studyThe pathogenesis of NAFLD and the therapeutic mechanism of Zhishi Daozhi Decoction are related to intestinal flora;The mechanism of action of Polyene phosphatidylcholine and Emodin may have nothing to do with intestinal flora;This experiment failed to demonstrate whether the mechanism of action of Berberine is related to intestinal flora. |
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