Experimental Study On The Protective Effects Of ZhuangTongYin On The Kidney Deficiency And Blood Stasis Syndrome Of Diabetic Nephropathy Rats’ Kidney

Objective: Through the observation of the effects of Chinese fomula Zhuang Tong Yin（ZTY）on the biochemical indexes of renal fuctions,the renal pathomorphology,the protein kinase C（PKC）activity of renal tissue,and the m RNA expression of Connective tissue growth factor（CTGF）,Plasminogen activator inhibitor^-1(PAI^-1),Fibronectin（FN）,Transforming growth factor-β₁（TGF-β₁）and other cytokines of renal cotex in the diabetic nephropathy（DN）rats with kidney deficiency and blood stasis syndrome,thus to explore the mechanisms of ZTY in protecting the kidney of the rats and provide scientific bases for the exploration of the Guangxi specific Zhuang Drugs in preventing and treating DN.Methods: The kidney deficiency and blood stasis syndrome of diabetic nephropathy rat models were induced and prepared by single kidney nephrectomy combined with injection of Streptozotocin（STZ）in fasting abdominal cavity,and all of the rats were randomly divided into 7 groups: blank control group,sham operation group,model group,western medicine group,ZTY low dose group,ZTY medium dose group and ZTY high dose group.The intragastric dose of the ZTY groups were 6.8 g·kg^-1·d^-1 for the low dose group,13.6g·kg^-1·d^-1 for the medium dose group and 27.2 g·kg^-1·d^-1 for the high dose group.The western medicine group was also the positive control group of which the intragastric dose was 1g·kg^-1·d^-1 of captopril.During the treatment process,the general conditions of the rats in each group were observed and the changes of their body weight（BW）and random blood glucose（RBG）were constantly monitored every 2 weeks.6 weeks later,the right kidney indexes（RKI）and fasting blood glucose（FBG）of rats in each group were tested;though full / semi automatic biochemical analyzer the urine,serum and the biochemical indexes of renal function of rats in each group were detected,such as the 24 h urinary protein,Creatinine clearance rate（Ccr）,Blood urea nitrogen（BUN）,β₂-Microglobulin（β₂-MG）and Cystatin C（Cys C）were detected;by optical microscopy and transmission electron microscope（TEM）the pathological changes of glomerulus of rats in each group were observed;though the immunohistochemical SABC staining method the protein kinase C（PKC）activity of rats renal tissues were detected;and by fluorescent quantitative Real Time-Polymerase Chain Reaction（RT-PCR）the m RNA expression of CTGF,PAI^-1,FN,TGF-β₁ cytokines in renal cortex were measured.What’s more,The data obtained were expressed as mean±standard deviation（（?）±S）,and the comparison between two groups were realized by statistical analysis of variance,meanwhile if P<0.05,the criterion for the difference has statistical significance was set.Results: The rats of model group were depressed,sluggish and huddled with the phenomenons like dry-yellow hair or alopecia,pale-clammy tail,dark lips and tongue,besides there also appeared the symptoms like polydipsia,polyphagia,polyuria,and emaciation;4 weeks after the treatment,all the above phenomenons and symptoms were improved in different degree.Comparing with the model group,the biochemical indexes of renal function of rats in each treatment group were decreased significantly（P<0.05）,such as 24 h urinary protein,Ccr,BUN,β₂-MG,Cys C（P<0.05）;the HE and PAS staining and the pathological changes under TEM were alleviated in varying degrees;all the PKC activities of rats renal tissues in treatment group were lowered（P<0.05）;all the rats renal m RNA expressions of CTGF,PAI^-1,FN,TGF-β₁ cytokines in treatment groups were reduced（P<0.05）;however,the comparisons between treatment groups have no statistical significance（P > 0.05）.Conclusions:（1）It shows that the rats in DN model group get the syndromes of kidney deficiency and blood stasis shows like depressed,sluggish,huddled,polydipsia,polyphagia,polyuria,etc..（2）The above detecting indexes suggest that ZTY can reduce the renal damage degree of DN rats with kidney deficiency and blood stasis syndrome,and play a certain therapeutic effect in delaying the progression of the disease.（3）The study suggest that the protective function of ZTY on kidney of DN rats with kidney deficiency and blood stasis syndrome is realized by various factors,such its the inhibition effects on PKC activities and m RNA expression of CTGF,PAI^-1,FN,TGF-β₁ cytokines though improving the five renal function and alleviating the pathological morphological changes.