| Purpose:Guided by Traditional Chinese Medicine Theory and based on animal experiment, the paper shows effect of TFA on proteinuria of rats with early diabetic nephropathy, and discusses the possible active molecular mechanism.Materials and Methodology:40male SD rats were divided into4groups:normal control group (NS), model group (DN), low TFA dose group (LTFA), high TFA dose group (HTFA); models were made by intraperitoneal injection of streptozotocin (STZ), and after successful model, rats were gavaged with high and low dose (50mg/kg/d and200mg/kg/d)of TFA carboxymethyl cellulose solution respectively (CMC,30mg/ml), and normal control group was gavaged with the same amount of CMC solution. When the successive administration was given for24weeks, rat urines in24hours were collected since the last administration, and24-hour urinary protein and urinary albumin of rats in each group were analyzed by automatic biochemistry analyzer, and urinary creatinine values were measured by elisa. When rental biopsy was performed, pathological changes of rental tissue were observed through electron microscope, HE staining and PAS staining. Expression of Nephrin protein level and mRNA of rental tissue were measured by protein immunoblotting and real-time PCR.Results:Experimental results showed that24h-UTP in DN group was significant higher than that in NS group, and the difference was statistically significant (P<0.05). After intervention of high and low dose of TFA in early diabetic nephropathy model,24-UTP and Urimary mAlb/Cr were all lower than that in DN group,but compared with DN group,only the former had significant difference (P<0.05); rental biopsy under electron microscope and HE staining showed that:incrassation of basement membrane, disordered arrangement and decreased number of podocyte, broadening and fusion of podocytic process in DN group. Both high and low dose of TFA intervention witnessed improvement of glomerular basement membrane lesions, reduction of extracellular matrix, increase of the number of podocyte and alleviation of podocytic process fusion; ratio of positive PAS staining area and tangent area of each glomerulus by image analysis software showed that ratio of rental tissue positive PAS staining area and tangent area of each glomerulus in DN group was significantly higher (P<0.05), and both high and low dose of TFA intervention reduce the ratio of rental tissue positive PAS staining area and tangent area of each glomerulus, but compared with DN group, the former had no significant difference (P>0.05) but the latter had significant difference (P<0.05); protein immunoblotting and real-time PCR of rental tissue showed that after low and high dose of TFA intervention in early diabetic nephropathy model, the expression of nephrin in rental tissue and its mRNA levels were obviously lower than that in DN group,but compared with DN group,only the later had significant difference(P<0.05).Conclusion:TFA can significantly reduce24h-UTP and Urinary mAlb/Cr of rats with early diabetic nephropathy as well as alleviate pathological injuries, which are likely realized through increasing the expression of Nephrin mRNA and protein in rental tissue podocytes. |
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