Preparation Of Compound Rhizoma Coptidis Capsule And Its Intervention Effect On Diabetic Nephropathy Rats

Objectives: The purpose of this study was to prepare Compound Rhizoma Coptidis capsules(CRCC)and control its quality,and investigate the intervention effect of CRCC on DN rats with diabetic nephropathy.Methods:Prepare of CRCC according to the preparation approval. Rhizoma coptidis, radix puerariae, loquat leaf were identifiesd by TLC, The content of berberine hydrochloride was determined by HPLC,Chromatographic conditions : the chromatographic column SinoChrom ODS- BP(250mm x 4.6 mm, 5 microns), mobile phase: 0.3% phosphoric acid and acetonitrile(72:28),to join 0.25 ml triethylamine per 100 ml, detection wavelength : 345 nm, column temperature :25 ℃, the flow rate : 1.0 ml/min, Injection Volume:20 μl. And water, the difference of filling quantity, disintegration time limit and microbial limit were examined by reference to the 2010 edition of China pharmacopoeia. DN model rats were established by injecting streptozotocin(STZ).The rats were randomly divided into seven groups: the normal group,the model group,the enalapril treatment group,the xiaoke pill treatment group and three CRCC treatment groups.They were orally administered once a day for five weeks.The fasting blood glucose(FBG),blood urea nitrogen(BUN),serum creatinine(Scr), Insulin(Ins), 24 h urinary protein(24h Upro) and24 h urinary microalbumin(24h UmAlb) were tested.The ultrastructural changes of renal tissue were observed by the electron microscope,the pathological changes in renal tissues were examined by optical microscopy. Immunohistochemical measures were used to detect the expressions of TGF-β1、Smad2/3、P-Smad2、Smad7、BMP-7 、Smad1/5 、 SnoN 、 CTGF and AGEs protein, and RT-PCR was used to detect TGF-β1mRNA and BMP-7 mRNA in renal tissues.Results: The TLC identification was special for Berberine hydrochloride in Rhizoma coptidis, puerarin in radix puerariae, ursolic acid in loquat leaf. berberine hydrochloride showed a good linear relationship in the range of 0.0625 ~ 2.0 μg, r = 0.9999. The average recovery was99.4%, and RSD was 0.75%.Water, the difference of filling quantity, disintegration time limit and microbial limit conformed to the requirements of the Chinapharmacopoeia.Compared with model group,BUN、Scr、Ins、24h Upro and 24 UmAlb levels decreased at different degrees in CRCC treatment groups; the abnormal pathomorphology and ultrastructural in renal tissue was improved;immunohistochemistry results showed that the expression of TGF-β1 、 P-smad2 、Smad2/3、CTGF and AGEs were reduced,while the expression of BMP-7,Smad1/5 and Smad7 increased in CRCC treatment groups;the expression of TGF-β1 m RNA were reduced,but the expression of BMP-7 mRNA had no obvious change in CRCC treatment groups.Conclusions:The method of CRCC quality control is reliable, stable,simple, rapid, reproducibility, specificity, which could be used for quality control of CRCC, and the preparation quality of CRCC conformed to the requirements of the Chinese pharmacopoeia. CRCC can improve the early renal function, delay the progression of chronic renal pathology.