| Objectives:1 Use the network pharmacology method to screen the effective active ingredients of Jiang Tang Xiao Zhi prescription,and predict its potential targets in the treatment of type 2 diabetes and improvement of islet β-cell dedifferentiation.2 To observe the effect of different doses of Jiang Tang Xiao Zhi prescription on the dedifferentiation of islet β cells in spontaneously diabetic db/db mice,and to further explore its mechanism.Methods:1 Obtain all the chemical components of Jiang Tang Xiao Zhi prescription from the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform(TCMSP),and use oral bioavailability(OB)≥ 30%and drug-likeness(DL)≥ 0.18 as the standard for screening effective active ingredients.Obtain the targets of effective active ingredients through the Targets Information function of the TCMSP platform,and use Uniprot database to standardize the target names.Using "type 2 diabetes" and "β-cell dedifferentiation" as search terms,the GeneCards database was searched to obtain disease-related targets.The target genes of type 2 diabetes,βcell dedifferentiation and Jiang Tang Xiao Zhi prescription were intersected,and drawn into a Venn diagram of "drug target" and "disease target".Use Cytoscape_v3.9.1 software to construct a visualized network of " Jiang Tang Xiao Zhi prescription-Drug-Active Component-TargetDisease".The drug-disease common target genes were imported into the String V.11.5 database to construct a protein interaction network,and the analysis results were imported into Cytoscape_v3.9.1 software in TSV format,and the network topology was analyzed using the Analyze Network function.GO and KEGG pathway enrichment analysis of drug-disease common targets were performed using Metascape database.2 Taking C57BL/6J mice as normal group,BKS-Lepr/Gpt(db/db)mice were randomly divided into model group(model),metformin group(metformin),Jiang Tang Xiao Zhi prescription low-dose group(JTXZL),Jiang Tang Xiao Zhi prescription medium dose group(JTXZM),Jiang Tang Xiao Zhi prescription high dose group(JTXZH).All mice were fed with normal maintenance feed for 8 weeks.All mice were fed with a drug volume of 1ml/100g body weight.The doses of Jiang Tang Xiao Zhi prescription low,medium and high dose groups were 2.5 g/kg/d,5.0 g/kg/d,10.0 g/kg/d respectively.The dose of metformin was 2.5 g/kg/d.All drugs were dissolved in normal saline;corresponding doses of normal saline were given to the blank control group and the model group.Gavage 1 time/day,continuous intervention for 8 weeks.The general condition,fasting blood sugar and body weight of the mice were observed.On the 8th week of administration,intraperitoneal injection of glucose tolerance test and insulin tolerance test was performed.After the 8th week of the experiment,samples were collected to detect serum FINS,TG,CHOL,HDL-C,LDL-C,and HE staining to observe pancreatic pathology;PCR to detect the expression level of Pdx1 mRNA,Ngn3 mRNA,FoxO1 mRNA;Western Blot to detect pancreatic protein expression of IGF1-PI3K-Akt signaling pathway and the protein expression levels of β cell maturation marker(Pdx1)and dedifferentiation into precursor cell marker(Ngn3).Results:1 The potential mechanism of Jiang Tang Xiao Zhi prescription to improve the dedifferentiation of islet β cells in type 2 diabetes is relatively complex,involving 46 active ingredients of traditional Chinese medicine,106 targets,4733 biological processes,418 cellular components,and 633 molecular functions,241 signal pathways.Among them,the PI3K/Akt signaling pathway and the FoxO signaling pathway are highly enriched,which may be related to the potential mechanism of Jiang Tang Xiao Zhi prescription in improving the dedifferentiation of islet β cells in type 2 diabetes.2 The results of animal experiments showed that,compared with control group,the body weight and the levers of FBG,FINS,HOMA-IR,TG,HDL-C,CHOL were all significantly increased;the pancreatic islets were significantly hypertrophied,and the vacuolar degeneration cells increased;the expressions of Pdx1 mRNA,Ngn3 mRNA,and FoxO1 mRNA tended to increase(P>0.05),and the expressions of Pdx1,Ngn3,IGF1,PI3K,and p-Akt proteins were significantly enhanced(P<0.05,P<0.01,P<0.001)in the model group.Compared with model group,the body weight of the mice in JTXZL group,JTXZM group,and JTXZH group had a tendency to decrease(P>0.05);the metformin group and all JTXZ groups had no significant improvement in FBG(P>0.05);the levers of FINS were significantly reduced in JTXZM group and JTXZH group(P<0.05);the HOMA-IR was significantly reduced in JTXZH group(P<0.05);metformin group had no significant improvement on FINS and HOMA-IR(P>0.05);the lever of TG Significantly decreased in JTXZH group(P<0.05),the levers of CHOL and LDL-C had a downward trend(P>0.05);pancreatic vacuolar degeneration cells in JTXZH group and metformin group were significantly reduced;the expressions of Pdxl mRNA,Ngn3 mRNA and FoxO1 mRNA tended to be down-regulated in metformin group,JTXZL group,JTXZM group and JTXZH group(P>0.05);the expression of Ngn3 in the metformin group,JTXZM group,and JTXZH group was significantly reduced(P<0.05,P<0.01);the expression of Pdxl in the metformin group was significantly reduced(P<0.05),and the expression of Pdxl in the JTXZH group showed a downward trend(P>0.05);the expressions of IGF 1 and PI3K were significantly weakened(P<0.05,P<0.01),and the expression of p-Akt tended to decrease(P>0.05)in the metformin and JTXZH groups.Compared with metformin group,the levers of FINS in JTXZM group and JTXZH group were significantly decreased(P<0.05);the HOMA-IR in JTXZH group was significantly decreased(P<0.05).Conclusion:1 Jiang Tang Xiao Zhi prescription can significantly improve insulin resistance and lipid metabolism in db/db mice.2 Jiang Tang Xiao Zhi prescription can improve the vacuolar degeneration cells of pancreas in db/db mice,and improve the dedifferentiation of islet β cells.3 Jiang Tang Xiao Zhi prescription may improve pancreatic β-cell dedifferentiation by inhibiting the IGF1/PI3K/Akt signaling pathway and down-regulating the expression of transcription factor FoxO1. |
PDF Full Text Request