ABSTRACT
A novel paper-based analytical device(PAD)was prepared and applied to determine the xanthine oxi-dase(XOD)inhibitory activity of Salvia miltiorrhiza extracts(SME).First,polycaprolactone was 3D printed on filter paper and heated to form hydrophobic barriers.Then the modified paper was cut according to the specific design.Necessary reagents including XOD for the colorimetric assay were immobilized on two separate pieces of paper.By simply adding phosphate buffer,the reaction was performed on the double-layer PAD.Quantitative results were obtained by analyzing the color intensity with the specialized device system(consisting of a smartphone,a detection box and sandwich plates).The 3D-printed detection box was small,with a size of 9.0 cm x 7.0 cm x 11.5 cm.Color component G performed well in terms of linearity and detection limits and thus was identified as the index.The reaction con-ditions were optimized using a definitive screening design.Moreover,a 10％glycerol solution was found to be a suitable stabilizer.When the stabilizer was added,the activity of XOD could be maintained for at least 15 days under 4℃or-20℃storage conditions.The inhibitory activity of SME was investigated and compared to that of allopurinol.The results obtained with the PAD showed agreement with those ob-tained with the microplate method.In conclusion,the proposed PAD method is simple,accurate and has a potential for point-of-care testing.It also holds promise for use in rapid quality testing of medicinal herbs,intermediate products,and preparations of traditional Chinese medicines.
ABSTRACT
OBJECTIVBE To investigate the intervention of compound Astragalus and Salvia milt-iorrhiza extract (CASE) consisted of astragalosides, astragalus polysaccharides and salvianolic acids on the interaction of microRNA-145/microRNA-21 (miR-145/miR-21) and Smad3C/3L phosphorylation (pSmad3C/pSmad3L) down-stream of transforming growth factor-β (TGF-β)/mitogen activated protein kinase (MAPK) signaling in hepatocellular carcinoma (HCC) progression by in vitro and in vivo experi-ments. METHODS In HepG2 cells and xenografts of nude mice, antagomir/agomir and plasmids of Smad3C/3L phosphorylation site mutation (Smad3 3S-A/Smad3 EPSM) were used to intervene miR-145/miR-21 and pSmad3C/pSmad3L expression respectively,then incorporative CASE treatment. Cell proliferation, migration, apoptosis, tumor growth and histopathologic characteristics of xenografts, relevant proteins of TGF-β/Smad pathway and miR-145/miR-21 were evaluated.RESULTS CASE up-regulated miR-145 while down-regulated miR-21, inhibited cell proliferation,migration and tumor growth, accelerated cell apoptosis in HepG2 cells respectively transfected with Smad3 WT, Smad3 EPSM,Smad3 3S-A plasmids in cultured dishes and xenografts of nude mice,the above effects were more evident in HepG2 cells with increased pSmad3C.In TGF-β1-stimulated HepG2 cells and xenografts of nude mice, CASE antagonized the facilitating effects of miR-145 antagomir/miR-21 agomir on cell migration,proliferation,tumor growth and inhibiting effects of miR-145 antagomir/miR-21 agomir on cell apoptosis; CASE increased miR-145 down-regulated by miR-145 antagomir and decreased miR-21 up-regulated by miR-21 agomir,reduced protein level of pSmad3L and their proteins including TβRⅡ, pERK1/2, pJNK1/2 and pp38 while elevated pSmad3C expression. CONCLUSION These results suggest that pSmad3C/pSmad3L maybe interact with miR-145/miR-21 in HCC progression,which may be one of important molecular mechanisms of CASE's anti-HCC effects.
ABSTRACT
Objective: To investigate spectrum-effect relationship of the extract from Salvia miltiorrhiza on HIF-1α of retinal Müller cells in the advanced glycationend products (AGEs) or hypoxia conditions, regarding tanshinone IIA as reference to explore the relationship between pharmacological effects and chemical substances, and to reveal the material basis of S. miltiorrhiza treating diabetic retinopathy (DR). Methods: Fingerprints of different S. miltiorrhiza extract were established by HPLC and characteristic peak's area was recorded. After that, the expression quantity of HIF-1α of retinal Müller cells were tested in the AGEs or hypoxia conditions under the two different conductions: S. miltiorrhiza extract or only tanshinone IIA. Gray relational analysis and partial least squares regression analysis (PLSR) were combined to build spectrum-effect relationship. Results: Seventeen characteristic peaks were marked out. Compared with the model set, 10 batches of extracts and tanshinone IIA could lower the expression quantity of HIF-1α. The statistical results showed that the peaks 1, 5, 6, 9, 10, 11, 12, 13, 14, 15, and 16 would inhibit HIF-1α expression, which made great contribution. Conclusion: The 15,16-dihydrotanshinone I, cryptotanshinone, tanshinone I, tanshinone IIA, and other six components might be the main effective components in the ethanol extract from S. miltiorrhiza.