| Objective: The purpose of this experiment is to study the intervention effect of Huayu and Tongluo traditional Chinese medicine on rats with diabetic kidney disease(DKD)and its effect on TGF-β1 / p38 MAPK pathway in the kidney in order to provide a way for clinical prevention and treatment of DKD.Methods: 60 healthy male SD rats were randomly divided into normal group(C group,n = 8)and modelling group(n = 52).And 52 rats in the modelling group were fed with high sugar and high fat diet,combined with streptozotocin(STZ)by intraperitoneal injection(35 mg/kg)to establish the model of diabetic rats.After 72 h,the random blood glucose of rats was measured,which was ≥16.7mmol/L was considered as successful model.The model rats were randomly divided into 4 groups: model group(M group),Huayu group(H group),Tongluo group(T group)and Irbesartan group(I group).All rats in treatment groups were fed with the corresponding drugs by gavage once a day for 16 weeks.Blood and urine samples were collected at the end of the 16 th week to detect 24 h urinary total protein(24h UTP)and related blood indexes.Renal tissues were taken to detect indexes of oxidative stress and observe the pathological changes of kidney.Immunohistochemistry technique(IHC),Real-time PCR and Western Blot were used to detect the expression of protein and mRNA of TGF-β1 / p38 MAPK signaling pathway.Results1 The general status of rats he rats in C group were in good condition;The rats in M group showed mental depression,polydrink,polydipsia,polyuria and emaciation;The rats in treatment groups were improved in different degrees.2 Comparison of 24 h urinary total protein(24h UTP)of rats ompared with C group,the 24 h UTP of rats in M group was significantly increased(P<0.01);Compared with M group,the 24 h UTP of rats in treatment groups was significantly decreased(P<0.01);Compared with I group,it was significantly increased in H group(P<0.05),and there was no difference between I and T group(P>0.05).3 Comparison of random blood glucose(RBG)and blood lipid of rats ompared with C group,the RBG of rats in M group was significantly increased(P<0.01);There was no statistical difference in RBG of rats among modelling groups(P>0.05).Compared with C group,the total cholesterol(TC),triglyceride(TG)and low density lipid(LDL)of rats in M group was increased(P< 0.01);Compared with M group,the blood lipid of rats in treatment groups was decreased(P<0.01);In terms of TC and LDL,there was no statistical difference among treatment groups(P>0.05);In terms of TG,compared with I group,it was significantly decreased in T group(P<0.05),and there was no statistical difference between I and H group(P>0.05).4 Comparison of renal function of rats Compared with C group,the blood urea nitrogen(BUN)and serum creatinine(Scr)and uric acid(UA)of rats was increased in M group(P<0.01);Compared with M group,the indexes of renal function of rats in treatment groups was decreased(P<0.05 or P<0.01);and there was no statistical difference among treatment groups(P>0.05).5 Comparison of oxidative stress indicators of rats Compared with C group,the activity of SOD were decreased,and the content of MDA were increased in M group(P<0.01);Compared with M group,the oxidative stress indicators of rats in treatment groups was improved(P<0.01);and there was no statistical difference among treatment groups(P>0.05).6 Morphological observation of kidney Rats in C group had normal renal structure.It appeared glomerular hypertrophy,narrow renal capsule,the accumulation of mesangial matrix,and diffuse thickening of glomerular basement membrane(GBM)in kidney of rats in M group;The results of electron microscope showed that the GBM was thickened,and the foot process appeared diffuse fusion and even disappeared in M group.The rats in treatment groups were improved in different degrees.7 Detection of the expression of TGF-β1 and p-p38 MAPK in renal tissue by immunohistochemistry(IHC)In the glomeruli,only a small amount of TGF-β1 and p-p38 MAPK were expressed in C group;And there was a large amount of expression in M group;Protein expression in treatment groups was reduced in different degrees than that in M group.8 Detection of the expression of TGF-β1 and p38 MAPK mRNA in renal tissue by Real-time PCR Compared with C group,the expression of TGF-β1 mRNA in M group was increased(P<0.01);Compared with M group,it was decreased in treatment groups(P<0.05 or P<0.01);and there was no statistical difference among treatment groups(P>0.05).There was no statistical difference among all of the groups in the expression of p38 MAPK mRNA(P>0.05).9 Detection of the protein expression of TGF-β1,p-p38 MAPK and p38 MAPK in renal tissue by Western Blot In terms of the expression of TGF-β1 and p-p38 mapk protein,compared with C group,it was increased in M group(P<0.01);Compared with M group,it was decreased in treatment groups(P<0.01);and there was no statistical difference among treatment groups(P>0.05).There was no statistical difference among all of the groups in the expression of p38MAPK(P>0.05).Conclusions:1.Huayu and Tongluo traditional Chinese medicine can improve the general condition of DKD rats,regulate blood lipid,decrease urinary protein,protect renal function,alleviate renal pathological damage,and have obvious protective effects on kidney of DKD rats.2.Both Huayu and Tongluo herbs can inhibit oxidative stress of kidney,and down-regulate the expression of TGF-β1 and p-p38 MAPK,suggesting that inhibiting the TGF-β1/p38 MAPK pathway through controlling oxidative stress may be one of the mechanisms of reducing ECM accumulation and its protective effect on the kidney of DKD rats.3.The effect of Tongluo traditional Chinese medicine on reducing urine protein is similar with irbesartan.4.The effect of Tongluo traditional Chinese medicine on reducing urine protein and TG is better than Huayu traditional Chinese medicine. |
