| Purpose:The potential targets and mechanism of action of diosgenin in T1 DM were explored by network pharmacology.The effects of diosgenin on islet β cells apoptosis index and Caspase-3 protein expression in T1 DM mice were further studied by establishing animal experimental models,so as to further explore the anti-apoptosis effect of diosgenin on islet βcells in T1 DM mice,providing new ideas and methods for the prevention and treatment of T1 DM with traditional Chinese medicine.Materials and methods:1.Network pharmacologyAfter the Smile pattern of diosgenin was retrieved in Pubchem,the action targets of diosgenin were predicted by Swiss Target Prediction,Super-PRE and Ch EMBL data platforms.Gene Cards and Dis Ge NET databases were used to predict the targets related to T1 DM,and the common targets of both were selected to construct the protein-protein and drug-disease interaction networks.Cytoscape3.8.2 software was used to visualize the "drug Active ingredient-disease target-pathway" and "Protein-protein interaction" network diagrams.2.Animal experiments:Eighty-nine male C57BL/6 mice aged 6-8 weeks and weighing 20-25 g were selected and fed adaptively for one week.Twelve mice were randomly selected as the normal control group,and the remaining 77 mice were used for modeling.After 12 hours of fasting without water,the mice were injected into the abdominal cavity with the concentration of 1% STZ solution(PH=4.2),the injection dose was 50mg/kg,and the mice were repeatedly injected for five consecutive days.Thus,the mice were induced into T1 DM model.72 hours later,the blood glucose of mice was detected by the tail vein sampling method.When the blood glucose value was greater than 16.7mmol/L,the experimental modeling was successful.Model mice were randomly divided into 4 groups with 12 mice in each group according to the random number table method.They were named as model group,diosgenin low-dose group,diosgenin high-dose group and prednisone group,respectively.Model group and each treatment group were given intragastric administration once a day until the end of the experiment at the 12 th week,during which the general state,blood sugar and body weight of mice were observed.The expression of Caspase-3 protein in mouse pancreas was detected by Western Blot.The apoptosis index of pancreatic β cells was detected by TUNEL method.During the experiment,all mice were fed ordinary growth and reproduction diet,drinking water was tap water,and were free to move,eat and drink.Results:1.Pharmacologic study on T1 DM network treated by diosgeninA total of 114 diosgenin targets and 1356 T1 DM related targets were obtained,67 diosgenin and T1 DM cross targets were obtained,and 12 key targets such as VEGFA,STAT3 and HIF1 A were obtained by PPI network screening.GO functional enrichment analysis showed that the biological processes involved in diosgenin target genes were mainly related to cell response to nitrogen compounds,hormone response,peptide response,positive regulation of cell migration,response to insulin,response to exogenous stimuli,inflammatory response and other processes.KEGG pathway enrichment analysis showed that the target was mainly enriched in PI3K-Akt signaling pathway,HIF-1 signaling pathway,insulin signaling pathway,etc.2.Animal experimental research2.1 Behavioral observation of mice in each group: the mental state of mice in the model group was poor,the fur color was dull,the body was thin,the reaction was slow,the intake of food and water increased,the polyuria,and the padding material needed to be changed frequently.Mice in the administration group also showed the above symptoms,but the symptoms were less severe and the body weight was heavier than that in the model group.2.2 Changes in blood sugar of mice in each group: compared with normal group,blood sugar of mice in each group was significantly increased(P < 0.05);Compared with model group,blood glucose in all groups was significantly decreased after 4 weeks of medication(P < 0.05),and the downward trend was more significant in high-dose group(P < 0.05).Compared with prednisone group,blood glucose of mice in low-dose and high-dose groups was significantly decreased(P < 0.05).2.3 Changes in body weight of mice in all groups: compared with mice in normal group,the body weight of mice in all groups was decreased(P < 0.05);Compared with model group,diosgenin low-dose group showed an increasing trend after 12 weeks(P < 0.05).The body weight of mice in high-dose group showed an increasing trend from 4 weeks(P < 0.05).Compared with prednisone group,the weight of mice in high-dose group increased more significantly(P < 0.05).2.4 Apoptosis index of mice islet beta cells in each group: The apoptosis of mice islet beta cells in each group was observed under a light microscope,and scattered apoptotic cells were observed in the normal group.Compared with normal group,the number of apoptosis of pancreatic β cells in model group,diosgenin high dose,low dose and prednisone groups was significantly increased(P < 0.05).Compared with model group,the number of apoptosis of pancreatic β cells in diosgenin low-dose,high-dose and prednisone groups was significantly decreased(P < 0.05),and the trend of apoptosis was more significant in high-dose groups.2.5 Caspase-3 protein changes of mice in each group: Compared with the normal group,the expression of Caspase-3 protein in the pancreas of mice in model group,diosgenin low-dose group,high-dose group and prednisone group was significantly higherConclusion:1.Diosgenin extract of Pangolin has 12 key targets with T1 DM,which can play a role in the treatment of T1 DM by participating in apoptosis,immunity,oxidative stress and other processes.2.Diosgenin extract of Dioscorea diosgenin can lower blood sugar and improve the general state of T1 DM in mice.3.Diosgenin extract of Pangolin can down-regulate Caspase-3 protein,reduce the apoptosis index of islet beta cells,and then play a role in protecting and repairing islet beta cells. |
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