| Purpose:Based on the AMPK/SIRT1/FOXO1 signal pathway,the effects of the combination of Huangqi Gegen Tang and Gualou Tang on endothelial function,glucose and lipid metabolism and oxidative stress in diabetic atherosclerotic rats were investigated,and the mechanism of the combination of Huangqi Gegen Tang and Gualou Tang in reducing blood glucose and lipid and alleviating aortic endothelial dysfunction in diabetic atherosclerotic rats was clarified,providing a theoretical basis for the development and clinical application of diabetic atherosclerotic drugs.Material and method: SPF grade SD rats(male,68 rats)were randomly divided into blank group(10 rats),model group(15 rats),positive control group(TMK group)(15 rats),Huangqi Gegen decoction and Gualou decoction combined decoction group(14 rats),Huangqi Gegen decoction and Gualou decoction combined granule group(14 rats).In addition to the blank group,each group was fed with high-fat diet for 28 days,and the experimental model of diabetes atherosclerosis was induced by intraperitoneal injection of 30 mg · kg-1 2% streptozotocin(STZ)on the29 th day.The corresponding drug or distilled water of 10 ml · kg-1 was injected into the stomach on the 3rd day of STZ,once a day,for 49 consecutive days.Blood glucose(Glu)was detected by blood glucose meter,triglyceride(TG),cholesterol(CHO),high-density lipoprotein(HDL-C),low-density lipoprotein(LDL-C)by biochemical analyzer,and propylene glycol(MDA),vascular hemophilia factor(vWF),nitric oxide(NO),reactive oxygen species(ROS),nitric oxide synthase(e NOS),superoxide dismutase(SOD),glycosylated hemoglobin(Hb A1c)by enzyme-linked immunosorbent assay(ELISA)The content of endothelin-1(ET-1),the relative expression of adiponectin receptor(Adipo R1),sterol regulatory element-binding protein-1c(SREBP-1c),silent information regulatory factor-1(SIRT1),adenylate-activated protein kinase(AMPK),glucose transporter 4(GLUT4)m RNA were detected by real-time fluorescence quantitative PCR.Western blot was used to detect silent information regulator 1(SIRT1),autophagy-associated protein(Beclin-1),forkhead box protein 01(FOXO1),and B-cell lymphoma factor 2(Bcl-2).The experimental data were analyzed and processed with statistical software SPSS26.0.Results:1.Glucose and lipid of atherosclerosis rats with diabetes(1)Effect on blood glucose of diabetes atherosclerotic rats: there was no difference in blood glucose between groups before modeling and 28 days after high-fat diet(P > 0.05);After 49 days of treatment,compared with the blank group,the blood glucose in the model group was higher(P<0.05).Compared with the model group,blood glucose in TMK group,decoction group and granule group decreased(P<0.05).(2)The effect on blood lipid of atherosclerosis rats with diabetes(1)Effect on serum CHO of diabetes atherosclerosis rats: CHO of model group rats was higher(P < 0.05).Compared with the model group,CHO in TMK group,decoction group and granule group decreased(P<0.05).(2)Effect on serum TG of diabetes atherosclerosis rats: TG of model group rats was higher(P<0.05).Compared with the model group,TG in TMK group,decoction group and granule group decreased(P < 0.05).(3)Effect on serum LDL-C of diabetes atherosclerosis rats:LDL-C of model group rats was higher(P<0.05).Compared with the model group,LDL-C in TMK group,decoction group and granule group were lower(P<0.05).(4)Effect on serum HDL-C of diabetes atherosclerosis rats: HDL-C of model group rats was lower(P < 0.05).Compared with the model group,HDL-C in TMK group,decoction group and granule group increased(P<0.05).2.Expression of SIRT1 m RNA and GLUT4 m RNA in aorta of atherosclerosis rats with diabetes(1)Effect on the expression of SIRT1 m RNA in the aorta of diabetes atherosclerotic rats: The expression of SIRT1 m RNA in the aorta of the model group was reduced,which was different from that of the blank group(P<0.05).Compared with model group,TMK group,decoction group and granule group could increase the expression of SIRT1 m RNA(P < 0.05).The effect of granule group was better than that of decoction group(P<0.05).(2)Effect on the expression of GLUT4 m RNA in aorta of diabetes atherosclerotic rats:The expression of GLUT4 m RNA in model group was lower than that in blank group(P<0.05).Compared with the model group,TMK group,decoction group and granule group could increase the expression of GLUT4 m RNA(P<0.05).There was no difference between the effects of granule and decoction groups(P>0.05).3.Expression of AMPK m RNA,Adipo R1 m RNA and SREBP-1c m RNA in the aorta and liver of atherosclerotic rats with diabetes(1)Effect on Adipo R1 m RNA expression in aorta of diabetes atherosclerotic rats:Adipo R1 m RNA expression in model group was lower than that in blank group(P<0.05).Compared with the model group,TMK group,decoction group and granule group could increase the expression of Adipo R1 m RNA(P < 0.05).There was no difference between the effect of granule and decoction groups(P>0.05).(2)Effect on the expression of AMPK m RNA in aorta of diabetes atherosclerotic rats:The AMPK m RNA expression in model group was lower than that in blank group(P<0.05).Compared with the model group,TMK group,decoction group and granule group could increase the expression of AMPK m RNA(P < 0.05).The effect of granule group was better than that of decoction group(P<0.05).(3)Effect on the expression of SREBP-1c m RNA in the liver of atherosclerotic rats with diabetes: The expression of SREBP-1c m RNA in the model group was increased compared with the blank group(P<0.05).Compared with the model group,TMK group,decoction group and granule group could reduce the expression of SREBP-1c m RNA(P < 0.05).The effect of granule group was better than that of decoction group(P<0.05).4.Contents of MDA,ROS,SOD,Hb A1 c in diabetes atherosclerotic rats(1)Effect on the content of serum MDA in diabetes atherosclerosis rats: MDA in model group rats increased(P < 0.05).Compared with the model group,MDA in TMK group,decoction group and granule group decreased(P<0.05).There was no difference between the effect of granule and decoction groups(P>0.05).(2)Effect on serum ROS content of diabetes atherosclerotic rats: ROS in model group was increased(P < 0.05).Compared with the model group,ROS in TMK group,decoction group and granule group decreased(P < 0.05).There was no difference between the effect of granule and decoction groups(P>0.05).(3)Effect on serum SOD content of diabetes atherosclerosis rats: SOD in model group rats was reduced(P<0.05).Compared with the model group,SOD in TMK group,decoction group and granule group was higher(P<0.05).There was no difference between the effect of granule and decoction groups(P>0.05).(4)Effect on serum Hb A1 c content of diabetes atherosclerosis rats: Hb A1 c content of model group rats increased(P<0.05).Compared with the model group,Hb A1 c in TMK group,decoction group and granule group decreased(P<0.05).The effect of granule group was better than that of decoction group(P<0.05).5.The content of e NOS,NO,ET-1,vWF in diabetes atherosclerotic rats(1)Effect on serum e NOS content of diabetes atherosclerosis rats: e NOS content of model group rats was reduced(P<0.05).Compared with the model group,the content of e NOS in TMK group,decoction group and granule group was increased(P<0.05).There was no difference between the effect of granule and decoction groups(P>0.05).(2)Effect on serum NO content of diabetes atherosclerosis rats: serum NO of model group rats was reduced(P<0.05).Compared with the model group,the content of NO in TMK group,decoction group and granule group was increased(P < 0.05).The effect of granule group was better than that of decoction group(P<0.05).(3)Effect on the content of serum ET-1 in diabetes atherosclerotic rats: The level of serum ET-1 in model group rats increased(P<0.05).Compared with the model group,the content of ET-1 in serum of rats in TMK group,decoction group and granule group decreased(P<0.05).There was no difference between the effect of granule and decoction groups(P>0.05).(4)Effect on serum vWF content of diabetes atherosclerosis rats: The serum vWF of model group rats was increased(P<0.05).Compared with the model group,the TMK group,the decoction group and the granule group decreased the serum vWF content of rats(P<0.05).There was no difference between the effect of granule and decoction groups(P>0.05).6.Expression of SIRT1,FOXO1,Bcl-2 and Beclin-1 proteins in aorta of diabetes atherosclerotic rats(1)Effect on the expression of SIRT1 protein in aorta of atherosclerosis rats with diabetes: The expression of SIRT1 protein in model group rats was reduced(P<0.05).Compared with the model group,the expression of SIRT1 protein was increased in TMK group,decoction group and granule group(P < 0.05).The effect of granule group was better than that of decoction group(P<0.05).(2)Effect on FOXO1 protein expression in aorta of atherosclerosis rats with diabetes:FOXO1 protein expression in model group rats was reduced(P<0.05).Compared with the model group,the expression of SIRT1 protein was increased in TMK group,decoction group and granule group(P<0.05).The effect of granule group was better than that of decoction group(P<0.05).(3)Effect on Bcl-2 protein expression in aorta of diabetes atherosclerotic rats: Bcl-2protein expression in model group was reduced(P<0.05).Compared with the model group,the expression of Bcl-2 protein was increased in TMK group,decoction group and granule group(P < 0.05).The effect of granule group was better than that of decoction group(P<0.05).(4)Effect on Beclin-1 protein expression in aorta of diabetes atherosclerotic rats:Beclin-1 protein expression in model group rats was increased(P<0.05).Compared with the model group,the expression of Beclin-1 protein in TMK group,decoction group and granule group decreased(P<0.05).The effect of granule group was better than that of decoction group(P<0.05).7.Histomorphology of aorta in atherosclerotic rats with diabetes After HE staining of rat aortic tissue,it was found that in the blank group,the structure of aortic vascular wall was clear,the endothelial cells were orderly,the morphology was clear and complete,the lipid nucleus was small,and no plaque was found.In the model group,the cells of aortic cells were in disorder,with vague shape,incomplete structure,thickened intima,a large number of foam cells,lipid crystals,and atherosclerotic plaques.Compared with the model group,the aortic cells in the TMK group were slightly disordered in arrangement,slightly fuzzy in shape,slightly complete in structure,with larger fat nuclei,foam cells visible,and a small amount of plaque formation.Aortic cells in the decoction group and granule group were orderly arranged,clear in shape,complete in structure,small in lipid nucleus,and atherosclerotic plaques were seen in some areas.Conclusion:1.The combination of Huangqi Gegen Decoction and Gualou Decoction and the combination of Huangqi Gegen Decoction and Gualou Decoction can play an effective role in reducing blood glucose and lipid,mainly by stimulating the secretion of Adipo R1,activating the SIRT1/AMPK signal pathway,and then regulating the activity of downstream factors GLUT4 and SREBP-1c,to reduce blood glucose and lipid in diabetic atherosclerotic rats.2.The combination of Huangqi Gegen Decoction and Gualou Decoction and the combination of Huangqi Gegen Decoction and Gualou Decoction can effectively reduce the oxidative stress reaction,mainly by activating the SIRT1/Mn SOD signal pathway,reducing the content of MDA,ROS,Hb A1 c,and increasing the content of SOD,to reduce the oxidative stress reaction in diabetic atherosclerosis rats.3.The combination of Huangqi Gegen decoction and Gualou decoction,Chinese herbal decoction and granules of Huangqi Gegen decoction and Gualou decoction can effectively alleviate endothelial dysfunction,mainly by activating the SIRT1/FOXO1 signal pathway,increasing Bcl-2,reducing the level of Beclin-1 protein,thereby inhibiting cell apoptosis,increasing cell autophagy,and improving endothelial dysfunction in diabetic atherosclerotic rats.4.The combination of Huangqi Gegen Decoction and Gualou Decoction and the combination of Huangqi Gegen Decoction and Gualou Decoction can effectively alleviate endothelial dysfunction,mainly by activating the SIRT1/e NOS signal pathway to increase the content of e NOS and NO,reduce the content of ET-1 and vWF,reduce the damage of vascular endothelial cells,and regulate the endothelial function of aorta in diabetic atherosclerotic rats. |
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