| Purpose:In this study,the comprehensive analysis of plant compound enzymes on body weight,water intake,fasting blood glucose,blood lipid levels,inflammatory factors,oxidative stress indicators and liver histopathology in type 2 diabetic rats was used to evaluate the hypoglycemic and lipid-lowering effects of plant compound enzymes.Through serum non-target metabolomics analysis,the hypoglycemic mechanism of plant compound enzymes was initially explored.The purpose is to use the theory of traditional Chinese medicine as a guide,select "medicine and food homologous" Chinese herbal medicines,and apply modern fermentation technology to prepare a plant compound enzyme functional food with hypoglycemic effect.In order to fully carry forward the theory of traditional Chinese medicine,apply it to basic research and guide the diagnosis and treatment of clinical diseases.Material and method:Thesis one:75 SPF Wistar male rats were selected,10 rats were randomly selected and included in the normal control group,fed with maintenance diet,and the remaining 65 rats were fed with high-fat diet.The animal model of type 2 diabetes was established by intraperitoneal injection of STZ.Type 2 diabetic rats were divided into 5 groups and given different intervention measures.The rats in the normal control group and the model control group were given 10 m L/kg/d of 0.9% normal saline by gavage,the rats in the traditional Chinese medicine group were given the traditional Chinese medicine compound Yitangkang by gavage at 10 m L/kg/d,and the rats in the western medicine group were given acarbose tablets.10 m L/kg/d was given by gavage,the raw material group was given 10 m L/kg/d of unfermented enzyme raw material,and the enzyme group was given 10 m L/kg/d of plant compound enzyme by gavage,once a day,for 8 weeks..The fasting blood glucose,body weight,and water intake of the rats were detected before modeling and at the 2nd,4th,6th,and 8th weekends after modeling,and the materials were collected at the 8th weekend.The content of TG,TC,LDL-C,HDL-C,AST and ALT in serum was determined by colorimetric method,and the content of inflammatory factors IL-6,TNF-α and antioxidant enzymes in serum were detected by enzyme-linked immunosorbent assay(ELISA).Related indexes SOD,GSH,CAT,MDA,HE staining were used to observe the pathological results of rat liver tissue.Thesis two:The experimental animals,feeding methods,grouping and intervention measures are the same as those of the first paper.Liquid chromatography-mass spectrometry(LC-MS)was used to analyze the changes of differential metabolites and metabolic pathways among the groups.Results:Thesis one:1.Changes in the general condition of the rats: the rats in the normal control group have always maintained a good mental state,with normal food intake,water intake,and stools,and their hair is shiny and smooth;The rats in the other groups were fed with high-fat diet,and after injection of STZ,each group had a typical "three to one" type 2 diabetes symptoms.After the intervention,compared with the model control group,the mental state of rats in each group was improved,the hair is brighter,the activity increased,and the symptoms of "three more and less" were alleviated.2.The results of body weight monitoring of rats showed that: before the intervention,compared with the normal control group,the body weights of the other five groups were significantly increased;after 2 and 4 weeks of intervention,the body weights of the model control group and each intervention group were significantly decreased;After 8 weeks,compared with the model control group,the body weights of the other four groups tended to be stable;during the experiment,the weight of the rats in the normal control group gradually increased,while the body weight of the rats in the model control group continued to decrease.The weight control of rats in traditional Chinese medicine group is the best,the results of Western medicine group and raw material group are similar,there is no significant difference,slightly worse than that in traditional Chinese medicine group,and the results of enzyme on weight control are slightly worse.3.The monitoring results of water consumption of rats showed that compared with the normal control group,the water consumption of the remaining five groups of rats increased significantly;during the 2nd and 4th weeks of the intervention,the water consumption of the rats in each group did not change significantly from that before the intervention;From the 6th week of intervention,compared with the model control group,the rats in the traditional Chinese medicine group drank less water;after 8 weeks of intervention,compared with the rats in the model control group,the increase trend of the rats in each intervention group was significantly slower.The curative effect of the group was the best,the enzyme group was slightly worse than the traditional Chinese medicine group,and the effect of the traditional Chinese medicine group and the enzyme group was significantly better than that of the western medicine group and the raw material group.4.Fasting blood glucose monitoring results showed that: compared with the normal control group,the FBG of the rest of the groups were significantly increased;during the intervention process,the blood glucose of the rats in the model control group always showed an upward trend,and the FBG of the rats in each intervention group was higher than that in the model group.Compared with the obvious decrease,the control level of FBG in the traditional Chinese medicine group and the western medicine group is always better than that in the raw material group and the enzyme group.5.The monitoring results of blood lipid levels in rats showed that compared with the normal control group,the blood lipid indexes of the rats in the model control group were abnormal,such as increased TC,TG,LDL-C,and decreased HDL-C.After the intervention in the experimental group,the levels of TC,TG and LDL-C in each group were significantly decreased,and the level of HDL-C was significantly increased;among them,the enzyme group had the best effect on reducing the levels of TC,TG and LDL-C in type 2 diabetic rats.The raw material group had the best effect on raising the level of HDL-C.6.The serum liver function test results of diabetic rats showed that compared with the normal control group,the serum AST and ALT levels of the model control group and the raw material group were significantly increased,and the other four groups had no statistical difference;compared with the model control group,the The levels of AST in the traditional Chinese medicine group,the western medicine group and the enzyme group were significantly decreased,and the levels of ALT in the serum of the rats were decreased in the traditional Chinese medicine group and the enzyme group.7.Changes in liver histomorphology: The arrangement of hepatocytes in the traditional Chinese medicine group,the enzyme group and the raw material group was relatively regular,and the pathological changes such as cell volume and lipid vacuoles were significantly reduced and alleviated,and the nuclear shift was improved in varying degrees.8.Expression of inflammatory factors in rat serum: Compared with the normal control group,except the Chinese medicine group,the other IL-6 levels were increased;compared with the model control group,the Chinese medicine group and the western medicine group significantly decreased IL-6-6 level,the difference in the enzyme group was not significant.Compared with the normal control group,the content of TNF-α in the model control group and the raw material group was significantly increased.After the intervention of each experimental group,the content of TNF-α in the traditional Chinese medicine group and the enzyme group was significantly decreased,and the other groups had no statistical difference.9.The detection results of serum oxidative stress indicators in rats showed that: compared with the normal control group,the SOD and CAT activities of the model control group decreased,the GSH level was significantly decreased,and the MDA activity was significantly increased;compared with the model control group,the traditional Chinese medicine group,The SOD activity of the rats in the enzyme group increased,the level of GSH in the western medicine group and the enzyme group increased,the MDA activity in each intervention group decreased significantly,and there was no statistical difference in the CAT activity in the intervention groups.Thesis two:1.Differential metabolite analysis: compared with the normal control group,a total of 385 differential serum metabolites(VIP>1 and p<0.05)were screened out as potential biomarkers for type 2 diabetes.There were 356 differential serum metabolites between the enzyme group and the model control group as potential biomarkers for the treatment of type 2 diabetes(VIP>1 and p<0.05).2.Differential metabolic pathway analysis: The results showed that in the comparison between the normal control group and the model control group,there were a total of 26 significantly different metabolic pathways,of which 7 metabolic pathways were up-regulated and 19 were down-regulated.In the enzyme group,there were a total of 27 differential metabolic pathways,of which 12 metabolic pathways were up-regulated and 15 were down-regulated.Conclusion:1.The glucose and lipid metabolism disorders and liver cell damage of type 2 diabetic rats were effectively improved by plant compound enzymes.2.The inflammatory response and oxidative stress response of type 2 diabetic rats were improved by plant compound enzymes.3.Plant compound enzymes can up-regulate amino acid metabolism,glucose metabolism and other metabolic signaling pathways to improve abnormal glucose metabolism in rats. |
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