Study On The Medication Rules Of Yitangkang In The Treatment Of Type 2 Diabetes And Its Mechanism Of Action On Intestinal Flora And Intestinal Inflammatio

Purpose:The medical record data mining method was selected to analyze the clinical case data of Professor Shi Yan’s treatment of type 2 diabetes through traditional Chinese medicine inheritance and calculation platform,Excel,SPSS and other software,and summarize the drug use law,aiming to provide traceable guidance for clinical treatment of type 2 diabetes.Then through animal experiments,we observed the effects of Yitangkang on intestinal flora,inflammatory factors and oxidative stress process,and analyzed and discussed the related mechanism of Yitangkang in treating type 2 diabetes.Material and method:1.Collect the clinical data of type 2 diabetes patients received by Professor Shi Yan in the expert clinic of the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine from September 2021 to March 2023,screen them according to the inclusion and exclusion criteria,standardize the name of traditional Chinese medicine,arrange the data,set up an independent database,upload it to the traditional Chinese medicine inheritance computing platform（V3.0）,and use the functions of"statistical analysis"and"prescription analysis"to conduct medication frequency,channel-tropism,Four Properties,Five Tastes and Efficacy Statistics;Import the prepared binary matrix data into the SPSS Modeler 18.0 software,analyze the association rules of high-frequency drugs according to the default parameters of the system according to the Apriori algorithm,and build the association rule network diagram of Chinese medicine for the treatment of type 2 diabetes;Through SPSS Statistics 25.0 software,systematic cluster analysis was carried out on high-frequency Chinese medicine,and systematic cluster pedigree diagram was drawn according to the results to form various cluster drug groups.Finally,all data results were analyzed,and Professor Shi Yan’s experience in the differentiation and treatment of type 2 diabetes was summarized and inherited.2.40 SPF grade db/db mice were randomly divided into model group,low dose group,medium dose group,high dose group and dagliflozin group,and were given common diet.Eight SPF grade db/m mice were used as blank groups and were fed a common diet.The traditional Chinese medicine compound Yitangkang consists of 12 medicines,including coptis chinensis,yellow cypress,astragalus,yellow essence,atractylodes and panax notoginseng.After 1 week of adaptive feeding,mice in the low-dose group of Yitangkang were administered 7g·kg^-1·d^-1with Yitangkang decoction,mice in the middle dose group of Yitangkang were given 14g·kg^-1·d^-1with Yitangkang decoction,mice in the high-dose group were administered 28g·kg^-1·d^-1with the decoction,mice in Dagliflozin were given 1.29mg·kg^-1·d^-1,dosing for a total of 8 weeks.The blank and model groups were given daily saline gavage.Body weight and fasting blood glucose（FBG）values were measured every two days.After 8 weeks,the mice were fasted for12h before animal material extraction and tissue isolation.The intestinal histopathological and morphological changes were detected by HE staining;gene sequencing of intestinal flora by 16S-r DNA high-throughput sequencing technology;the expression level of intestinal ROS,SOD and LPS was measured by enzyme-linked immunosorbent assay（ELISA）;immunohistochemistry was used to detect intestinal IL-6 and IL-1βexpression levels;co-localization of claudin 1 and occludin in intestinal tissues was observed by immunofluorescent staining;the expression levels of TLR4 and NF-κB m RNA in intestinal tissues were detected by quantitative real-time PCR（q RT-PCR）;the protein expression levels of TLR4 and NF-κB were determined by immunoblot（Western blot）.Results:Data Mining Results:1.Medication situation:76 prescriptions of type 2 diabetes patients were collected this time,and 102 kinds of traditional Chinese medicines were used in total.35 medicines were used more than or equal to 5 times,of which the top 5 herbs used most frequently were licorice,astragalus,polygonatum,coptis chinensis,and salvia miltiorrhiza.Among the four herbs in traditional Chinese medicine,warm medicine is the main ingredient,while among the five herbs,sweet and bitter herbs are used the most frequently,far higher than other herbs.The medicine involves 12 meridians,mostly referring to the spleen,lung,heart,and stomach meridians.A total of 17 types of traditional Chinese medicine are used,with the main functions of tonifying deficiency,clearing heat,and promoting blood circulation and resolving blood stasis.2.Correlation analysis results:Perform association rule analysis on high-frequency drugs.When the minimum conditional support is 30%,the minimum rule confidence is set to100%,and the maximum number of antecedents is set to 1,8 association rule results that meet the conditions are obtained.The top 5 compatible drug pairs with the highest overall support are Astragalus membranaceus-Salvia miltiorrhiza,solomon’s seal-Atractylodes macrocephala,Astragalus membranaceus-Atractylodes macrocephala,solomon’s seal-red ginseng,Astragalus membranaceus-red ginseng.3.Cluster analysis results:Professor Shi Yan treated type 2 diabetes with five groups of high-frequency clustering:prescription 1:keel,oyster,wild jujube kernel,cinnamon twig,allium macrostemon,notoginseng,leech,angelica sinensis,and chuanxiong;Prescription 2:Red peony and Achyranthes bidentata;Recipe 3:Chinese yam,Cornus officinalis,Laotou grass,goji berries,and Scutellaria baicalensis;Recipe 4:Chaihu,Xiangfu,Fructus Aurantii,Rhubarb,Ginseng,Alisma orientalis;Recipe 5:Xuanshen,Ophiopogon japonicus,Danshen,Atractylodes macrocephala,Huangjing,Astragalus membranaceus,Red ginseng,Poria cocos,Huanglian,Kudzu root,Mulberry white bark,Schisandra chinensis,Licorice.Animal experiment results:1.General condition of mice:During the experiment,all the mice in the blank group were in good mental state,with bright fur,normal diet and water intake,and normal urine volume.In the model group,the mental state was depressed,the fur was yellow and rough,the body was dull,the diet and water intake increased significantly,the amount of urine was increased,and the body size of the mice increased significantly.The low dose group,medium dose group,high dose group,and dagliflozin group had better mental state than the model group,and improved than before,the fur color and gloss,diet volume and urine volume decreased to different degrees.2.Body weight of the mice:Compared with the blank group,the model,low,medium,high,and daglifloznet increased（P<0.01）,and the high and daglifloznet mice decreased after 8 weeks compared with the model group（P<0.05）.3.FBG condition of mice:Compared with the blank group,the FBG values of mice in the model group,the low dose group,the medium dose group,the high dose group,and the daggligin group were significantly increased after 8 weeks of intervention（P<0.01）.Compared with the model group,the FBG value of mice in the low dose group significantly decreased after 8 weeks of intervention（P<0.05）,the FBG values of mice in the middle dose group,high dose group,and daggligin group were significantly reduced after 8 weeks of intervention（P<0.01）.4.Intestinal histopathology:The colon tissue in the blank group was normal,the intestinal gland cells were rehearsed and intact,the structural villi were intact,and no pathological phenomena such as tissue edema,necrosis,and inflammatory cell infiltration were seen.Compared with the blank group,the model group mice had inflammatory tissue infiltration,cell swelling,intestinal crypt abscess,necrosis,disorder of intestinal glands,and erosion in the mucosal layer.Compared with the model group,the inflammatory cells in the Dagliflozin group and the high-dose group were significantly reduced,and the mucosal results were significantly clear;in the low-dose group,the colonic group had mild edema,and no obvious ulcer was observed.Compared with the model group,each treatment group showed different degrees of improvement,and the treatment effect was the best in the high dose group and dagliflozin group.5.Gene sequencing of the intestinal microflora:Compared with the blank group,the structure and composition of intestinal microflora of mice in the model group changed,the level of Bacteroidetes decreased significantly（P<0.05）,the proportion of Firmicutes and Proteobacteria increased（P<0.05）,and the ratio of Firmicutes/Bacteroidetes（F/B）increased（P<0.05）;Compared with the model group,the structure and composition of intestinal microflora of mice in the high dose group of Yitangkang changed,the levels of Firmicutes and Proteobacteria decreased significantly（P<0.05）,the proportion of Bacteroidetes increased（P<0.05）,and the ratio of Firmicutes/Bacteroidetes（F/B）decreased（P<0.05）.6.The intestinal ROS,SOD and LPS levels in mice:Compared with the blank group,the ROS and LPS in the intestinal tissues of the model group were significantly increased（P<0.01）and SOD were significantly lower（P<0.01）;compared with the model group,the ROS in the intestinal tissues of the high-dose and dagliflozin groups was significantly lower（P<0.05）,LPS and SOD increased（P<0.01）.7.The intestinal expression levels of IL-6 and IL-1βin mice:Compared with the blank group,the levels of IL-6 and IL-1βin the intestinal tissue of the model group mice were significantly increased（P<0.01）.Compared with the model group,the levels of IL-6 and IL-1βin the intestinal tissues of mice in the high-dose group and Daggligin group were significantly reduced（P<0.01）.8.Col-alization of claudin 1 and occludin:Compared with the blank group,the fluorescence intensity of claudin 1 and occludin of each mouse intestinal tissue model group was significantly decreased（P<0.01）;compared with the model group,the fluorescence intensity of claudin 1 and occludin was significantly increased（P<0.01）.9.Expression levels of TLR4 and NF-κB m RNA in the mouse intestine:Compared with the blank group of mice,the levels of TLR4 and NF-κB m RNA in model tissues were significantly increased（P<0.05）;compared with the model group mice,the TLR4 and NF-κB m RNA level in the intestinal tissue of the high dose group mice was significantly reduced（P<0.05）,the level of TLR4 m RNA in the intestinal tissue of mice treated with daggliptin was significantly reduced（P<0.01）,the level of NF-κB m RNA in the intestinal tissue of mice treated with dagglizin significantly decreased（P<0.05）.10.Protein expression levels of TLR4 and NF-κB in the mouse intestine:TLR4 and NF-κB protein levels were significantly higher in mice in the model group（P<0.01）,and TLR4 and NF-κB protein levels were significantly decreased in the high-dose and dagliflozin group compared with the model group（P<0.01）.Conclusion:1.The statistical analysis of Professor Shi Yan’s prescription rules for treating type 2diabetes reflects the academic thought of"treating from the spleen".He believes that the occurrence of diabetes is based on yin deficiency or qi deficiency,marked by turbid phlegm and blood stasis,and most of them are caused by deficiency and excess.Blood stasis runs through diabetes all the time,and the core treatment method is"invigorating the spleen and benefiting qi,nourishing yin,clearing heat and activating blood".2.Professor Shi Yan treats type 2 diabetes from the starting point of"internal heat due to yin deficiency and loss of movement due to spleen deficiency".He uses more sweet drugs to replenish spleen and qi,and bitter drugs to clear heat.He is good at using tonifying deficiency drugs in combination with heat clearing drugs,blood activating and stasis removing drugs,and treats diabetes based on syndrome differentiation and symptoms.3.The traditional Chinese medicine compound Yitangkang can reduce the blood glucose of db/db mice,improve the body weight,mental state,fur color gloss,reduce the diet volume,water intake and urine volume of db/db mice.4.The traditional Chinese medicine compound Yitangkang may regulate intestinal flora,alleviate intestinal pathological changes in db/db mice,improve damaged intestinal mucosal barrier,alleviate peroxidation,and alleviate inflammatory reactions in their intestines.