Based On VEGF/PI3K/Akt Signaling Pathway To Explore The Mechanism Of Enlistment Of Astragalus And Angelica Pairs To Interfere With Renal Tissue Fibrosis Of Diabetic Kidney Disease Rats

Objective:To investigate the effect of astragalus membranaceus combined with angelica sinensis on the related biochemical indexes and urinary protein excretion rate of diabetic kidney disease(DKD)rats by regulating the level of mRNA related to VEGF/PI3K/Akt pathway.The expression of VEGF/PI3K/Akt pathway mRNA protein was detected by immunohistochemistry and quantitative Real-time PCR(RT-PCR),in order to explore the mechanism of astragalus and angelica medicinal pairs on VEGF/PI3K/Akt pathway.To provide experimental basis for clinical treatment of diabetic nephropathy with astragalus and angelica medicinal pairs.Methods:Experimental study 1.Streptozocin(STZ)30mg/kg was injected intraperitoneally to establish DKD rat model.Rats were divided into model group,astragalus and angelica medicinal pairs group(astragalus: angelicae sinensis = 3:2,5.40g/kg·d)and dapagliflozin group(0.45mg/kg·d),with5 rats in each group.Five SD rats of the same age were used as the blank control group.2.The body weight,urine volume,urine glucose,urine ketone,and insulin were measured during administration.Glycated hemoglobin(Hb A1c)was measured before administration and after the last administration.3.After8 weeks,3 kidneys from each group were sacrificed,and paraffin sections of renal tissue were made according to the histopathological data for pathological and ultrastructure observation.The mRNA expressions of VEGF,PI3 K and Akt in renal tissue of DKD rats were detected by immunohistochemistry.The expressions of VEGF,PI3 K and Akt proteins were detected by RT-PCR,The experimental results of each group were statistically analyzed to explore the effect of astragalus and angelica medicinal pairs on the expression of VEGF/PI3K/Akt pathway mRNA protein on the process of renal fibrosis in DKD rats.Results:1.Effects on the symptoms and signs of DKD rats:(1)The autonomic activity of rats in the control group was normal.(2)The rats in the standard model group showed slow response.(3)The general condition of rats in the astragalus and angelica medicinal pairs group and dapagliflozin group were better than that in the standard model group.2.Effects on the body weight of DKD rats:(1)Before gavage,the body weight of rats in the standard model group,the astragalus and angelica medicinal pairs group and the dapagliflozin group increased faster than that in the control group(P <0.05).(2)After 8 weeks of gavage,the body weight of the astragalus and angelica medicinal pairs group and dapagliflozin groups increased significantly compared with the standard model group(P <0.05).(3)The body weight of rats in the astragalus and angelica medicinal pairs group increased more than that in the dapagliflozin treatment group.3.Effects on serum of DKD rats:(1)The levels of fasting plasma glucose(FPG),blood urea nitrogen(BUN),serum creatinine(Scr),total cholesterol(TC),triglycerides(TG),glycated serum protein(GSP)and Hb A1 c in the standard model group were significantly higher than those in the control group(P <0.05).(2)The levels of Hb A1 c,FPG,TC,TG,GSP,BUN and Scr in the astragalus and angelica medicinal pairs group and dapagliflozin group were significantly lower than those in the standard model group.(3)Dapagliflozin treatment group significantly reduced the levels of TG,GSP and BUN(P <0.01).(4)The levels of insulin(inspiration,INS)in the astragalus and angelica medicinal pairs group and dapagliflozin treatment groups were significantly lower than those in the control group(P <0.05),and the dapagliflozin group had a greater reduction(P <0.05).4.Effect on microscale albuminuria(MAU)and β2-microglobulin(β2-MG)in DKD rats:(1)The levels of β2-MG and MAU in the astragalus and angelica medicinal pairs group and dapagliflozin treatment group were significantly higher than those in the control group(P <0.01).(2)Compared with the standard model group,the levels of β2-MG and MAU in the astragalus and angelica medicinal pairs group and dapagliflozin treatment groups were significantly decreased,and the reduction in dapagliflozin treatment group was greater(P <0.05).5.The effect on renal histopathology in DKD rats:(1)The renal tubular epithelial cells in the standard model group showed significant granular changes compared with the control group and the glomerular cystic cavity had sclerotic changes.There were significantly proliferation of mesangial cells in the capillaries,significantly thickened basement membrane,and a small amount of granulocyte infiltration.(2)In the astragalus and angelica medicinal pairs group,the glomerular capillaries were clearer than those in the standard model group,and a small amount of renal tubular epithelial cells also had vacuolization.In the dapagliflozin treatment group,a small amount of renal tubular dilatation and granulocyte infiltration were also observed.6.The effect on renal fibrosis in DKD rats:(1)More blue-stained collagen fibers were observed in the interstitium and around blood vessels in the standard model group than in the control group.(2)A large number of blue collagen fibers were found in the local renal tubulointerstitium in both the astragalusand angelica medicinal pairs group and dapagliflozin group.7.The expression of VEGF/PI3K/Akt signaling pathway mRNA in renal tissue of DKD rats:(1)The mRNA levels of VEGF,PI3 K and Akt in the kidney tissue of the standard model group were lower than those in the control group(P <0.01).(2)The mRNA expression levels of VEGF,PI3 K and Akt in the astragalus and angelica medicinal pairs group and dapagliflozin treatment groups were significantly higher than those in the standard model group(P <0.05).(3)Compared with the dapagliflozin treatment group,the mRNA expression of VEGF,Akt,PI3 K from high to low.(4)The mRNA expression of VEGF in dapagliflozin group was slightly higher than that in astragalus and angelica medicinal pairs group(P <0.01).(5)The mRNA expression of Akt in the astragalus and angelica medicinal pairs group was slightly better than that in the dapagliflozin treatment group.(6)The expression of PI3 K mRNA between the astragalus and angelica medicinal pairs group and dapagliflozin treatment groups have no obvious differences.8.The effect on the protein expression of VEGF/PI3K/Akt signaling pathway in renal tissue of DKD rats:(1)The protein expression levels of VEGF,PI3 K and Akt in the standard model group were significantly lower than those in the control group(P <0.05).(2)The protein expressions of VEGF,PI3 K and Akt in the astragalus and angelica medicinal pairs group and dapagliflozin treatment groups were significantly higher than those in the standard model group(P <0.01).Conclusions:1.The astragalus and angelica medicinal pairs can improve the clinical symptoms and detection indicators of DKD rats,improve the quality of life,improve the blood glucose level,reduce the excretion of urinary protein,and restore partial renal function.2.Astragalus and angelica medicinal pairs can improve renal fibrosis,protect renal tissue and delay the development of DKD by interfering with the expression of VEGF/PI3K/Akt pathway related target mRNA protein.