Study On The Mechanism Of Danzhi Jiangtang Capsule On NOTCH Pathway Of Kidney Tissue In The Rat Model Of Early Diabetic Nephropatht

Objective:1.To observe the expression of Jagged1,Notch 1,Hes1 and hey1 protein of Notch signal pathway in kidney of early DN model rats with deficiency of Qi and Yin and stagnation of blood stasis,and to explore the correlation between early DN with deficiency of Qi and Yin and stagnation of blood stasis and Notch signal transduction pathway.2.To observe the effect of Yiqi Yangyin Huoxue traditional Chinese medicine Danzhi Jiangtang capsule on the Jagged1,Notch1,Hes1 of Notch signaling pathway and the expression of Hey1 protein in kidney tissue of rats with early DN Qi-yin deficiency and blood stasis syndrome,and to explore the mechanism of TCM in preventing and treating early Qi-yin deficiency and blood stasis syndrome.Methods:Male SD rats were used as experimental animals.After one week of adaptive feeding,normal group(K Group)and model group(M group)were randomly divided.In the model group,the left kidney was excised and fed with high sugar and high fat(10%lard,2.5% cholesterol,20% sucrose)diet for 4 weeks.After fasting for 12 hours,STZ was injected intraperitoneally(with 0.1 mmol / L citric acid buffer of ph4.2 as 2%concentration)according to 30 mg / kg body weight.Blood glucose was measured by tail vein blood sampling after 72 hours mmol / L,and the amount of urine,diet and drinking water increased significantly.It was a successful model.2 weeks later,it was confirmed as early DN model by urine albumin test.The normal group was administrated with proper amount of normal saline,the model group was administrated with Fructus aurantii,aconite,Qingpi Decoction and the rat model of deficiency of Qi and Yin with stasis induced by low temperature regulation.The digital random method after successful modeling was divided into model group(M),western medicine group(X),high dose group(G),middle dose group(Z)and low dose group(D).After 8weeks of treatment,blood and urine samples were collected,blood glucose(Glu),Serum creatinine(Scr),24-hour urinary microalbumin,kidney weight / body weight were measured,renal tissues were taken,pathological sections were made,hematoxylin eosin(HE)staining was performed,pathological changes of renal tissues were observed,Notch/Jagged1,Notch1,Hes1 and Hey1 protein expression were detected by immunohistochemistry,and Notch/Hes1-m RNA protein expression was detected by real-time fluorescence quantitative PCR.Results:(1)Change trend of blood glucose and serum creatinine: after 2 weeks of modeling,compared with the K,the blood glucose and serum creatinine values of the model group,the traditional Chinese medicine group and the western medicine group increased significantly(P<0.05);after 8 weeks of medication,compared with the M,the blood glucose and serum creatinine value of each dose of traditional Chinese medicine group decreased(P<0.05).(2)Change trend of 24-hour urinary microalbumin: compared with the K,the M increased significantly(P<0.05);compared with the M,the X and each dose group of traditional Chinese medicine decreased(P<0.05);compared with the X,the Z and the D increased(P<0.05);compared with the G,the M and the D increased(P<0.05).(3)Kidney index: compared with the K,the kidney weight ratio of the M increased significantly(P<0.05);compared with the M,the X and each dose group of traditional Chinese medicine decreased(P<0.05);compared with the X,each dose group of traditional Chinese Medicine decreased(P<0.05);compared with the Z,the G decreased(P<0.05),and the D increased(P<0.05).(4)Pathological observation of HE staining showed glomerular sclerosis,atrophy,mesangial matrix increase,mesangial thickening and widening,and nodular hyperplasia in the model group;the high and medium dose groups of traditional Chinese medicine had positive improvement on the above pathological changes.(5)Notch/Jagged1,Notch1,Hes1 and Hey1 protein expression:Jagged1:compared with K group,there was no significant difference in M group(P>0.05),but the average Jagged1 renal tissue content in the group was higher than that in the group,and the average of renal tissue content in the G group was lower than that in the group.Notch1:compared with K group,there was no statistical significance in the M group compared with the K group(P>0.05);however,the average value of the M group was higher than that of the K group,and the average value of the G group was lower than that of the group.Hes1:compared with K group,mean value of the M group was significantly higher than that of the K group(P<0.05);compared with the M group,the treatment group was significantly lower(P<0.05),but there was no difference between the treatment groups.Hey1:compared with K group,there was no significant difference between the group and the K group(P>0.05).Compared with the M group,the mean value of the X group was significantly higher(P<0.05),and the other treatment groups were not statistically significant(P>0.05).(6)Notch/Hes1-m RNA expression determination: compared with the normal value group,the model group increased significantly(P<0.05);compared with the model group,the western medicine group,the traditional Chinese medicine each dose group decreased significantly(P<0.05).Conclusions:Yiqi yangyin huoxue traditional chinese medicine Danzhi Jiangtang Capsule has a hypoglycemic effect on the early DN of qi-yin deficiency and blood stasis syndrome rat model.At the same time,by reducing the expression of Hes1 protein and m RNA in rat kidney tissue,play a protective role in renal function,reduce the formation of proteinuria,delay the development of DN.