RESUMO
Ligustrazine (LIG) is a purified and chemically identified component of the Chinese herb Ligusticum wallichii Franchat. It is a potent blocker of vasoconstriction and has strong scavenger of oxygen free radicals. We investigated the effect of LIG on renal tubulointerstitial fibrosis using a rat model of unilateral ureteral obstruction. Ligustrazine treatment significantly reduced the scores of interstitial collagen deposition, amounts of hydroxyproline, the density of myofibroblasts and macrophages, and amounts of transforming growth factor-ß1 (TGF-ß1) and connective tissue growth factor (CTGF) compared with their level in a saline-treated control group. Using quantitative polymerase chain reaction we found that LIG treatment significantly reduced the mRNA expression of TGF-ß1, CTGF, monocyte chemoattractant protein-1 and osteopontin. Moreover, the mRNA expression of hepatocyte growth factor and bone morphogenetic protein-7 were significantly increased by LIG. In vitro, LIG inhibited the TGF-ß1-induced loss of cytokeratin-18 expression and de novo increase of the expression of α-smooth muscle actin of HK-2 cells in a dose-dependent manner, which suggested that LIG could restrain the process of epithelial-myofibroblast transition of tubular epithelial cells. This study indicates that LIG can attenuate renal tubulointerstitial fibrosis. It might be useful as a potential candidate in the treatment of chronic renal diseases.
Assuntos
Túbulos Renais/patologia , Ligusticum/química , Pirazinas/farmacologia , Obstrução Ureteral/complicações , Vasodilatadores/farmacologia , Actinas/metabolismo , Animais , Proteína Morfogenética Óssea 7/genética , Linhagem Celular , Colágeno/metabolismo , Fator de Crescimento do Tecido Conjuntivo/genética , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Fibrose/etiologia , Fibrose/patologia , Fibrose/prevenção & controle , Humanos , Hidroxiprolina/metabolismo , Queratina-18/genética , Túbulos Renais/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Masculino , Modelos Animais , Miofibroblastos/efeitos dos fármacos , RNA Mensageiro/genética , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Obstrução Ureteral/patologiaRESUMO
A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed and validated for determining tacrolimus (FK506) in rat tissues to study the effect of Schisandra sphenanthera extract on FK506 tissue distribution. After a liquid-liquid extraction with ethyl acetate, FK506 and ascomycin (IS) were subjected to LC-MS/MS analysis using positive electrospray ionization under multiple reactions monitoring mode. Chromatographic separation of FK506 and ascomycin was achieved on a Hypersil BDS C(18) column with a mobile phase consisting of methanol-water (containing 2 mM ammonium acetate, 95 : 5, v/v). The intra- and inter-batch precision of the method were less than 8.8 and 9.8%, respectively. The intra- and inter-batch accuracies ranged from 97.5 to 104.0%. The lowest limit of quantification for FK506 was 0.5 ng/mL. The method was applied to a FK506 tissue distribution study with or without a dose of Wuzhi (WZ) tablet. Most of the FK506 tissue concentrations were slightly increased after a concomitant WZ tablet dose, but the whole blood concentration of FK506 was dramatically increased 3-fold after a concomitant WZ tablet dose. These results indicated that the LC-MS/MS method was rapid and sensitive enough to quantify FK506 in different rat tissues, and strict drug monitoring is recommended when co-administering WZ tablet in clinical use.