| Objective:Using Type 2 diabetes model(T2DM)to evaluate the effects of different doses of soy isoflavones on fasting blood glucose,body weight,food intake,water intake,blood indicators,liver,kidney,and pancreas in T2 DM mice.Methods:Seventy three-week-old healthy C57BL/6J male mice were randomly divided into normal mice(10)and type 2 diabetes model mice(60).Normal mice were fed with 10%fat for energy in the control group for maintenance feed.Type 2 diabetic model mice were fed with custom feed of 60% fat,20% carbohydrate,and 20% protein for 6 weeks.Both groups were given purified water..After 6 weeks,type 2 diabetes model mice were injected with a medium dose(80mg/kg)of streptozotocin twice,and normal mice were injected with the same sodium citrate buffer.After 5 days,the model mice were fasted for12 hours,and the tail tip blood was collected to detect fasting blood glucose.Fasting blood glucose ≥ 11.1mmol/L is a successful model.A total of 54 mice were successfully modeled,with a success rate of 90%.Subsequently,the type 2 diabetes model mice were randomly divided into type 2 diabetes model control group(10),metformin positive control group(10),low-dose soy isoflavone group(75mg/kg)(10),middle-dose group(150mg/kg)(10 animals),high-dose group(300mg/kg)(10 animals).Monitor water intake,food intake,fasting blood sugar and body weight every week.After 6 weeks of intragastric administration,the "four items" of blood lipids in mice were measured-triglycerides,cholesterol,high-density lipoprotein cholesterol(HDL),low density lipoprotein cholesterol(LDL),and serum Interleukin-1 β,liver,kidney and pancreas hypertrophy index,HE staining to observe the pathological changes of liver,kidney and pancreas.Results:1.Enrollment of experimental animalsDuring the experiment,two mice died in the T2 DM group,one died in the PC group,and one died in the SI group.In order to unify the number of each group,each group was fed 8 mice,a total of 48 mice.2.Comparison of fasting blood glucose levels in miceFasting blood glucose in NC group was significantly lower than T2 DM group,SI150 group(P<0.01),significantly lower than SI75 group(P<0.05),but there was no significant difference compared with SI300 group and PC group(P>0.05).3.Changes in mouse body weightThe intervention of soy isoflavones showed an overall increase in the weight of mice,and the increase in the high-dose group was significantly higher in the lower-dose group(p<0.05).4.Changes in mouse food intakeThere was no significant difference in food intake between the normal control group and the high-dose soybean isoflavone group(P>0.05).5.Changes in the amount of water the mice drinkThe water consumption of the NC group was significantly lower than that of the T2 DM group,SI75,SI150,and SI300 groups(P<0.01);the water consumption of the T2 DM group was significantly higher than that of the SI300 group(P<0.05),but there was no significant difference compared with the SI75 and SI150 groups(P>0.05).6.HE staining to observe the effect of soybean isoflavones on liver weight index and pathological changes in miceThe liver weight index of the NC group was significantly lower than that of the other groups(P<0.01).Compared with the T2 DM group,the liver weight index of the soybean isoflavone intervention group was reduced(P<0.01).There was no significant difference in liver weight index between the T2 DM group and the PC group(P>0.05).In the NC group,there were extensive degeneration of hepatocytes in the central vein,around the portal area and in the liver parenchyma,and the cytoplasm was loose and slightly stained and granular,and no obvious inflammatory changes were seen.In the T2 DM group,hepatocyte granular degeneration was widely seen around the central vein,the portal area and the liver parenchyma,and the cytoplasm was loose and slightly stained and granular;occasionally lymphocyte infiltration(blue arrow)was seen around the blood vessels.In the SI75 group,hepatocyte granular degeneration was widely seen around the central vein,portal area and liver parenchyma,and the cytoplasm was loose and slightly stained and granular;lymphocytes were occasionally infiltrated around blood vessels.In the SI150 group,granular degeneration of hepatocytes was more common in the central vein,around the portal area and in the liver parenchyma,and the cytoplasm was lightly stained and granular;focal lymphocyte infiltration was rare in the lobules.In the SI300 group,hepatocyte granular degeneration was widely seen around the central vein,the portal area and in the liver parenchyma,and the cytoplasm was lightly stained and granular;no obvious inflammatory changes were seen.7.HE staining to observe the effect of soy isoflavones on the morphological and pathological changes of the kidney tissue in experimental miceThe kidney weight index of the NC group was significantly lower than that of the T2 DM group,PC group,and SI75 group(P<0.01),and significantly lower than that of the SI150 group and SI300 group(P<0.05).There was no significant difference in renal weight index between the T2 DM group and the PC group,SI75,and SI150 groups(P>0.05),but the difference was extremely significant compared with the SI300 group(P<0.01).There was no statistically significant difference in renal weight index between the PC group and SI75 and SI150(P>0.05),but it was statistically significant compared with the SI300 group(P<0.05).The kidney weight index of the SI75 group was statistically significant compared with the SI300 group(P<0.05),but it was not statistically significant compared with the SI150 group(P>0.05).The kidney weight index of the SI150 group was not statistically significant compared with that of the SI300group(P>0.05).Histopathological sections in the NC group showed uniform distribution of glomeruli,uniform cell number and matrix in glomeruli,tightly arranged renal tubules,and no obvious interstitial proliferation;no obvious inflammatory changes were seen.In the T2 DM group,a small amount of renal tubular epithelial cells were degenerated with vacuoles,and small round vacuoles were seen in the cytoplasm;lymphocytes were infiltrated locally around the blood vessels.In the SI75 group,a small amount of renal tubular epithelial cell vacuolar degeneration,small circular vacuoles can be seen in the cytoplasm;lymphocytes focal infiltration can be seen around the local blood vessels.In the SI150 group,a small amount of renal tubular epithelial cells were vacuolated,and small round vacuoles were seen in the cytoplasm;no obvious inflammatory changes were seen.In the SI300 group,a small amount of renal tubular epithelial cell vacuolar degeneration,small round vacuoles were seen in the cytoplasm;no obvious inflammatory changes were seen.8.HE staining to observe the effect of soybean isoflavones on pancreatic index and pathological changes in miceThere was no statistically significant difference in pancreatic weight index among the groups(P>0.05).Tissue sections showed that the morphology of the pancreatic islets in the NC group was normal,the number of acinar cells was abundant,the division was clear,the top was eosinophilic,the base was basophilic,and the cell morphology was normal;there was no obvious inflammation in the tissue.In the T2 DM group,there were no islets in the field of view,and no other obvious abnormalities were seen.There was no obvious abnormality in the SI75 group.A small amount of acinar cell infiltration(black arrow)was seen in the local pancreatic islets in the SI150 group.There was no obvious abnormality in the SI300 group.9.The effect of soybean isoflavones on blood lipids in experimental mice9.1 Effect of Soy Isoflavones on Triglycerides of Experimental MiceThe TG level of SI300 group was significantly lower than that of NC group,T2 DM group and PC group(P<0.01).There was no statistically significant difference among the other groups(P>0.05).9.2 Effect of Soy Isoflavones on Cholesterol in Experimental MiceThe cholesterol level of T2 DM group was significantly higher than that of SI75group(P<0.05).The PC group was significantly higher than the SI300 group(P<0.01).There was no statistical difference in cholesterol levels among the remaining groups(P>0.05).9.3 The effect of soybean isoflavones on high-density lipoprotein cholesterol in experimental miceThere was no statistical difference in the changes of high-density lipoprotein cholesterol among the groups(P>0.05).9.4 Effect of Soy Isoflavones on Low Density Lipoprotein Cholesterol in Experimental MiceThe level of LDL cholesterol in the T2 DM group was significantly higher than that of the SI300 group and the PC group(P<0.05),the PC group was significantly lower than the SI75 group(P<0.01),and the SI75 group was significantly higher than the SI150 and SI300 groups(P<0.01).10.The effect of soybean isoflavones on the serum inflammatory factor interleukin-1β in miceCompared with the T2 DM group,the SI300 group can significantly reduce the level of inflammatory factor IL-1β in type 2 diabetic mice(P<0.05).11.Correlation analysis of blood lipid indexes and inflammatory factorsAnalyzing the correlation of each group,the value is positive,TG and IL-1β show a positive correlation,and IL-1β increases with the increase of TG content.The rest is not relevant.Conclusion:High-dose soybean isoflavone intervention for 6 weeks can effectively reduce the levels of fasting blood glucose,TG,LDL-C,and inflammatory factor-IL-1β in T2 DM model mice,and improve the symptoms of "polyphagia" and "polydipsia" in type 2diabetic mice.Relatively gain weight.It may also protect the liver and kidneys by reducing inflammation. |
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