| Objective To observe how Yangyin Huoxue Recipe affects growth factor (TGF-beta 1) , PA/PAI-1 system, and extracellular matrix metabolism regulation process in sodium depletion rat model of chronic cyclosporine A nephrotoxicity. To reveals the links of Yangyin Huoxue Recipe anti-fibrosis in protein and gene transcription. Method 50 SPF grade SD rats are randomly divided into three groups, 10 in each group: enalapril group, the low-dose group of Chinese medicine, medium-dose group of Chinese medicine, high-dose group of Chinese medicine. To establish rat model of chronic cyclosporine A nephrotoxicity, each group keeps low-salt diet, injected CsA subcutaneously with doses of 12.5mg/kg.d, simultaneously fed with distilled water, enalapril, low-dose, dose and high dose Chinese medicine. 24-hour urine is collected at the ist,2nd,3rd,4th weekend to test urine volume blood,urine creatinine, in calculating creatinine clearance. At 4th weekend all rats are harvested. The left kidney is taken to RT-PCR detection of renal interstitial TGF-beta, uPA, tPA, PAI-1, FN, typeⅣcollagen mRNA expression; The right kidney is taken to histological examination and immunohistochemical detection of TGF-beta 1, fibronectin, typeⅣcollagen expression. Results is analyzed with statistical methods of ONE WAY ANOVA analysis,T-test and figures. Result The rats creatinine clearance rate, 24-hour urine protein in the first two weekends deteriorate to peak, in addition to the blank group continue to deteriorate, and others improve in the group. At 4th weekend renal function of enalapril group, the low-dose group of Chinese medicine, medium-dose group of Chinese medicine, and high-dose group of Chinese medicine improved significantly, creatinine clearance rate increased (P＜0.05) and 24-hour urinary protein (P＞0.01) of Chinese medicine groups in a dose-dependent manner; With HE staining pathology, we can see in blank group some kidney tubular atrophy, tubular epithelial cell degeneration and necrosis obvious infiltration with inflammatory cells and collagen fibers hyperplasia. The remaining enalapril group, the low-dose group of Chinese medicine, medium-dose group of Chinese medicine, and high-dose group of Chinese medicine are weakened change in order. In Masson stain Glomerular and tubular around fibroelastosis can be seen in the blank group, enalapril group, and the low-dose group of Chinese medicine, especially blank Group is the most obvious; Immunohistochemical staining of the renal interstitiaiTGF-beta 1, FN, IV collagen are expressed in each group, and around renal tubule TGF-beta 1, FN, IV collagen expression is obvious, in which the blank group is the most significantly, but the expression reduce in those rats which fed with enalapril and Chinese medicine. PCR suggested in enalapril group and high-dose group of Chinese medicine rats' renal interstitial TGF-beta mRNA express with a minimum strength, which followed by blank group, the low-dose group of Chinese medicine and medium-dose group of Chinese medicine; The low-dose group of Chinese medicine, medium-dose group of Chinese medicine, and high-dose group of Chinese medicine are weakened. mRNA expression of UPA, tPA, PAI-1 seem not too much different between different groups. Conclusion Enalapril and Yangyin Huoxue Recipe improves the the deterioration of proteinuria and renal function caused by chronic cyclosporine A nephrotoxicity, and Yangyin Huoxue Recipe on the treatment of proteinuria is in a dose-dependent manner; The both anti-fibrosis effect may be associated with down-regulate the expression of renal interstitial TGF-beta 1 mRNA induced by CsA, which thereby blocks the the interstitial extracellular matrix synthesis caused by the TGF-beta, but how Yangyin Huoxue Recipe affects PA/PAI-1 systems is not yet clear in this experiment. |
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