Effects Of Lamps Kidney Granule On TGF-Smad Transduction Pathway In Rats With Diabetic Nephropathy

Objective: To observe the Erigeron Yishen granule(the original "Yunnan breviscpini capsule")effect on TGF-Smad pathway in diabetic nephropathy rats,to explore the role of targets and mechanisms of Yishen Granule on diabetic nephropathy at protein level,to provide experimental basis for clinical treatment of diabetic kidney disease.Methods: 1.Animal grouping: clean and healthy adult male SD rats,weight(230±20)g,age of 8 weeks.Methods: 90 rats were randomly divided into normal group(n= A)and streptozotocin group(n = 75)in the model group(n = 15).The successful model of the rats were randomly divided into model group(B group),Benner Pury hydrochloride group(C group)and Erigeron Yishen granule low dose group(D group),group of Yishen granule(group E)and Erigeron Yishen granule high dose group(F group)15.2.Model preparation: B group,C group,D group,E group and F group with 1%citrate buffer solution of streptozotocin(STZ,Streptozotocin),according to 55mg/kg in rats on the left side of one-time intraperitoneal injection,A group with normal saline intraperitoneal injection.After modeling for 72 hours,the blood glucose value of caudal vein was > 3 16.7mmol/L,and the model was successfully prepared.Fed diabetic rats after 1 weeks of random blood glucose than 16.7mmol/L and urine volume larger than that of the control group,50% glucose was strongly positive,24 h urinary albumin was higher than that of control group 50% successfully for diabetic nephropathy model.3.Packet delivery: A group and B group were treated by saline 10ml/(kg.d),Cgroup received intragastric administration;Benner Pury hydrochloride 1.05mg/(kg.d),D group,gavage;group E,group F were treated with Erigeron Yishen granule 1.89g/(kg.d),9.45g/(kg.d),18.9g/(kg.d),intragastric gavage for 6 weeks.4.The index observation: the rats in each group were weighed once a week;in the stomach before the end of the sixth week,rats were taken to check the urine of rats in each group;the second day,said the weight,abdominal aorta blood,centrifugal to be tested.A multifunctional automatic biochemical detection of cystatin C(Cystatin C Cys C)Serum Creatinine Scr),serum creatinine,blood urea nitrogen(Blood Uria nitrogen BUN),urine protein / urine creatinine(Urine protein/Urine creatinine Up/Ucr)and other biochemical indicators;the kidneys,weighing,determination of expression of transport in renal tissues of rats with TGF-beta 1 by immunohistochemical method Smad2,Smad7,collagen,fibronectin.Image analysis with pathological image analysis system.The positive rate of TGF-1,Smad2,Smad7,type IV collagen,FN positive staining in renal tissue was detected with the optional visual field in 5 eyes.Statistical analysis based on computer image analysis data.Results: 1.Detection of biochemical indicators:(1)Fasting blood glucose,kidney index(kidney weight / body weight),urine protein / urine creatinine(Up/Ucr),Cystatin C,urea nitrogen,compared with A group and B group,the difference was statistically significant((P < 0.01).(2)Compared to Up/Ucr Cys,kidney index,C: and B group,C group and D group decreased Up/Ucr ratio and Cys content of C(P<0.05),E group,high dose group of kidney decreased significantly(P<0.01);compared with C group,E group decreased,Erigeron Yishen high dose group,the ratio of Up/Ucr the content of C Cys(P<0.05);compared with D group,E group and F group decreased Up/Ucr ratio and Cys content of C(P<0.05).(3)Compared with B group,the BUN content of C group,D group and F group were decreased(P<0.05),E group was significantly lower(P<0.01);compared with the C group,the BUN content of E group was lower(P<0.05);the content of BUN in E group was lower than that in D group(P<0.05).Compared with E group,the BUN content of F group increased(P<0.05).(4)There was no significant difference in Scr: between the two groups(P>0.05).2.Immunohistochemistry TGF-beta 1,Smad2,Smad7,collagen,FN detection:(1)Compared with B group,C group,D group,the expression of TGF-in renal tissue of Smad2,beta 1,type IV collagen,FN protein decreased(P<0.05);E group and F group were significantly decreased(P<0.01);Compared with C group,E group,the expression of F in kidney tissue of group TGF-beta 1,Smad2,type IV collagen,decrease of FN protein(P<0.05).(2)Compared with B group,the expression of Smad7 protein in C group and D group was increased(P<0.05),E group and F group were significantly increased(P<0.01).Compared with C group,the expression of Smad7 in renal tissue of E group and F group increased(P<0.05).Conclusion: The basic condition of the DN rats can be improved by the combination of breviscapine and Benner Pury,both of which can reduce the excretion of urine microalbumin and renal injury.Erigeron Yishen granule 3 dose group,middle dose group and high dose group had better effects of Yishen granule,in high dose group than Benner Pury hydrochloride group;immunohistochemical analysis indicated that breviscapine Yishen granule and Benner Pury hydrochloride can inhibit the activation of the TGF-Smad signaling pathway,reduce the expression of renal fibrosis.The effect of medium dose and high dose group was better than that of low dose group and Benner Pury group,and the therapeutic effect showed a dose effect relationship.