| [Objective] Taking type 2 diabetes mellitus with eczema as the target disease,the preparation method of Yalian gel was explored,and the potential target and mechanism of action of Coptis chinensis in the treatment of type 2 diabetes mellitus with eczema were predicted based on network pharmacology.Taking rats with type 2diabetes mellitus complicated with eczema as the research object,to explore and evaluate the efficacy and mechanism of Yalian gel in intervening type 2 diabetes mellitus complicated with eczema;At the same time,the safety and effect of Yalian gel in the treatment of type 2 diabetes mellitus with eczema were verified clinically,so as to fill the blank of topical Chinese medicine in the clinical treatment of type 2 diabetes mellitus with eczema.[Methods]1.Preparation of Yalian gel: To explore the extraction method of Yalian concentrate and preparation method of gel matrix,and to lay a foundation for the development of Yalian gel for the treatment of type 2 diabetes mellitus with eczema through the preparation process,quality standard,oral toxicity test and skin irritation test.2.Network pharmacology: the target genes of Coptis chinensis drugs were obtained from relevant databases to predict their possible molecular targets.The "diseasetarget" information of type 2 diabetes and eczema was obtained,and diseaseassociated targets were obtained.Related molecular biological analysis was conducted to explore the potential target and mechanism of rhizoma coptidis in the treatment of type 2 diabetes with eczema,and to screen the key target proteins.3.Animal experiments: with 10 Sprague wrote dawley rats as blank control group(K),ordinary feed and adequate drinking water.Another 60 SD rats,high-fat high-sugar diet after two weeks,the early mature building method to made type 2 diabetes mellitus rats by intraperitoneal injection of STZ.After the formation of the model,high fat and high sugar diet was continued to be fed and corresponding drugs were used for hypoglycemic intervention.On the first day of the ninth week,SD rats with type 2 diabetes mellitus complicated with eczema were made according to the method of animal model building.The successful SD male rats were randomly divided into model group(M),gel matrix group(J),mometasone furoate ointm group(Y),low-dose group(D),medium-dose group(Z)and high-dose group(G).The general state(body weight,food intake,water intake)of rats was observed daily.The fasting blood glucose level was detected on the morning of the first day of each intervention week.After 12 weeks of drug intervention,the rats were sacrificed,and blood samples were collected from abdominal aorta.Serum VCAM-1,SICAM,TNF-?,interleukin-1?(IL-1?)and interleukin-6(IL-6)were detected by ELISA.Skin tissue was collected and HE staining was performed to observe the pathological changes of skin tissue.RAGE,NF-?B and p47-phox were detected by real-time fluorescence quantitative(q PCR).RAGE,NF-?B and p47-phox were detected by Western blot(WB).4.Clinical trials: adopt randomized controlled research methods,in October 2019 to October 2020 in Hospital of Chengdu University of Traditional Chinese Medicine,inpatient and outpatient collection of type 2 diabetes mellitus and eczema(damp heat immersion type)cases,and in accordance with the principle of random grouping(check random number table)is divided into control group and treatment group: the control group on the basis of routine hypoglycemic external use western medicine treatment,the treatment group on the basis of routine hypoglycemic external use,even the Yalian gel treatment,a course of 2 weeks.[Results]1.Preparation results of Yalian gelThis preparation through the study of literature,selected the suitable gel base,and through the relevant experiment,the proportion of configuration,selected card,940 at2%,distilled water and triethanolamine,according to certain proportion and preparation technology of gel matrix,two times by reflux extraction and enrichment,have 1 g/ml of the concentration of the crude drug and even the concentrate.After mixing with the prepared gel matrix,a gelatin agent was obtained.The stability,toxicity and skin irritation were verified by the preliminary stability test and the safety test.2.Results of network pharmacological studies(1)Eleven potential active compounds of rhizoma coptidis chinensis were screened out from the database,corresponding to 90 target proteins;A total of 1580 "disease-targets" of type 2 diabetes mellitus with eczema were identified.After the association analysis of "drug-target" and "disease-target",a total of 47 co-acting targets of rhizoma coptidis chinensis and target diseases were obtained.Among them,the interleukin family,nuclear factor kappa B protein family and adhesion molecular immunoglobulin family are the core target groups,which are closely related to oxidative stress,inflammation and other pathways.(2)Through KEGG pathway enrichment analysis,a total of 97 pathways of rhizoma coptis for the treatment of type 2 diabetes combined with eczema were obtained.(3)network pharmacology research analysis,according to the results of rhizoma coptidis mechanism,prevention and control of type 2 diabetes mellitus and eczema may by changing the "coptis-type 2 diabetes mellitus and eczema" the expression of target genes of overlap,improve the level of RAGE,inhibiting inflammatory cytokines and the expression of adhesion molecules,regulation which-RAGE signal pathway,partly block RAGE in related to oxidative stress,inflammatory cytokines and adhesion molecules of the activated state for a long time.It is of great significance to improve the severity of eczema in patients with type 2 diabetes and provides information on possible drug targets for intervention.3.Results of animal experiments(1)Establishment of type 12 diabetes mellitus rat model and its effect on blood glucose(1)Under general conditions,compared with the blank group,the model group showed insensitive activity,decreased body weight,dark fur,increased water intake and food intake;(2)The fasting blood glucose index in model group was significantly increased(P <0.05).(3)After metformin intervention treatment,the body weight,mental state and fasting blood glucose of rats in model group were improved.Model of type 2 diabetic rats was established successfully.(2)Establishment of type 2 diabetes mellitus complicated with eczema in rats and the influence of related indexes(1)Compared with the blank group,the skin condition of rats in each model group was worse,with increased skin lesions,frequent scratching,and increased water consumption;(2)Objective indicators showed that fasting blood glucose,body weight and water intake were significantly increased in model group;(3)The skin lesions and general conditions were improved after the intervention of Yalian gel.(3)The effect of Yalian gel on serum related cytokines in ratsCompared with blank group,serum cytokines IL-6,IL-1?,TNF-?,VCAM-1 and SICAM in model group were significantly increased,and the difference was statistically significant(P<0.05).After experimental drug intervention,all serum indexes of rats showed a downward trend,and the difference was statistically significant(P <0.05).(4)Effects of Yalian gel on related regulatory factors in AGE-RAGE signaling pathway in ratsCompared with blank group,relevant regulatory factors in AGE-RAGE signaling pathway in skin tissue of rats in model group were significantly increased,with statistical difference(P < 0.01).Compared with model group,the m RNA and protein expressions of various related factors in skin tissue of rats after drug intervention showed a decreasing trend,and the differences were statistically significant(P < 0.05).4.Results of clinical trialsA total of 56 cases were included,of which 2 cases fell off and 1 case was excluded.53 cases were actually completed,including 26 cases in the control group and 27 cases in the treatment group.Baseline data such as gender and age of patients in the two groups(P > 0.05)were comparable.(1)Before and after treatment,there was a statistically significant difference in FPG between the control group and the treatment group(P<0.05);(2)After treatment,compared with the control group,EASI index score,pruritus index score,TCM subsymptom score,DLQI score,SAS self-rating anxiety scale score and SDS self-rating depression scale score all decreased,with statistical significance(P <0.05);[Conclusion]1.The preparation process of Yalian gel was finished.The stability,toxicity and skin irritation were verified by the preliminary stability test and the safety test.2.Rhizoma coptidis has the function characteristics of "multi-component,multitarget and multi-pathway",which can play a role in the prevention and treatment of type 2 diabetes mellitus complicated with eczema by regulating multiple target proteins and signaling pathways.3.Yalian gel has the effect of improving eczema symptoms,oxidative stress and inflammation in rats with type 2 diabetes mellitus complicated with eczema.To a certain extent,Yalian gel can improve the symptoms of polydipsia,polyphagia,weight loss and the pathological morphology of eczema in type 2 diabetes mellitus rats with eczema,and promote the recovery of eczema lesions through "AGERAGE signaling pathway".4.The elegant even the gels of treatment in patients with type 2 diabetes mellitus and eczema(hot and humid immersion type),a reduced FPG,dosing EASI index score before and after the reduction,dosing itch index score before and after the reduction,to reduce the disease of medicine of traditional Chinese medicine(TCM)before and after the time integral,reduced DLQI score before and after drug,drug delivery,SDS,SAS anxiety self-assessment scale before and after the depression self rating scale score.The results showed that Yalian gel was efficacy and safety in the treatment of patients with type 2 diabetes mellitus complicated with eczema(damp heat immersion type). |
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