Salvianolate inhibits reactive oxygen species production in H(2)O(2)-treated mouse cardiomyocytes in vitro via the TGFß pathway.

AIM: To investigate the effects of salvianolate, a water-soluble active compound from Salvia miltiorrhiza Bunge, on reactive oxygen species (ROS) production in mouse cardiomyocytes in vitro. METHODS: Primary ventricular cardiomyocytes were prepared from neonatal mouse. The cell viability was determined using MTT assay. Culture medium for each treatment was collected for measuring the levels of NO, iNOS, total antioxidant capacity (TAOC) and transforming growth factor ß1 (TGFß1). TGFß1 and Smad2/3 expression in the cells was detected with Western blotting. RESULTS: H2O2 (1.25 mmol/L) did not significantly affect the cell viability, whereas the high concentration of salvianolate (5 g/L) alone dramatically suppressed the cell viability. Treatment of the cells with H2O2 (1.25 mmol/L) markedly increased ROS and iNOS production, and decreased the levels of NO, TAOC and TGFß1 in the culture medium. Furthermore, the H2O2 treatment significantly increased TGFß1 and Smad2/3 expression in the cells. Addition of salvianolate (0.05, 0.1, and 0.5 g/L) concentration-dependently reversed the H2O2-induced alterations in the culture medium; addition of salvianolate (0.05 g/L) reversed the H2O2-induced increases of TGFß1 and Smad2/3 expression in the cells. Blockage of TGFß1 with its antibody (1 mg/L) abolished the above mentioned effects of salvianolate. CONCLUSION: Salvianolate inhibits ROS and iNOS production and increases TAOC and NO levels in H2O2-treated cardiomyocytes in vitro via downregulation of Smad2/3 and TGFß1 expression. High concentration of salvianolate causes cytotoxicity in mouse cardiomyocytes.

[Study on the fingerprint of processed Rhizoma Atractylodis Macrocephalae by HPLC].

OBJECTIVE: To establish HPLC fingerprint for the quality control of processed Rhizoma Atractylodis Macrocephalae (PRAM). METHODS: 14 batches of PRAM were collected from different places and were analyzed with the developed HPLC fingerprints method. The HPLC separation was performed on a Kromasil C18 analytical column (250 mm x 4.6 mm, 5 microm), and gradient elution was performed by mobile phase containing acetonitrile and water. The flow rate was 1.0 mL/min and the detection wavelength was at 242 nm. The temperature of column was 25 degrees C. RESULTS: Sixteen mutual peaks were selected in chromatography. Among the obtained fingerprints, the most of the detected peaks were separated effectively. The methodological evaluation showed that the method had a good repeatability. CONCLUSION: The RSD of relative retention time of mutual peaks which existed in all samples was less than 1.1%. The results of peak areas were in accordance with the request of fingerprint. The established fingerprint can be used for the quality control and species identifying of PRAM.