ABSTRACT
Objective To investigate the effect of SABP, a water-soluble component of Salvia miltiorrhiza, on the growth of orthotopic transplantation of H22 liver cancer and the immune microenvironment of liver cancer. Methods We established a mouse model of orthotopic transplantation of H22 cell liver cancer in BALB/c mice. ELISA was used to detect the expression of PD-L1, TGF-β, IL-1β, IL-10, IL-4, IFN-γ, IL-18, IL-7, IL-2, CCL-2 and CCL-21 in the liver. We counted the organ indexes of liver, spleen and kidney. Results SABP inhibited the growth of orthotopic transplantation tumors of H22 cell liver cancer, and increased the expression levels of PD-L1, TGF-β, IL-1β and IL-10 in the microenvironment of liver cancer, as well as the liver, spleen and kidney coefficients. Conclusion SABP could inhibit the growth of orthotopic transplantation tumors of H22 cell liver cancer and promote the expression of PD-L1, TGF-β, IL-1β and IL-10 in the microenvironment of liver cancer.
ABSTRACT
Objective::To establish an LC-MS/MS method for the determination of alcohol-soluble components and water-soluble components in Arnebiae Radix(L-shikonin, β, β′-dimethylacrylshikonin, β-acetoxy-isovalerylshikonin, lithospermic acid, caffeic acid, rosmarinic acid), for the purpose of quality control of the herb. Method::Chromatographic separation was carried out at 35 ℃ on Agilent ZORBAX SB C18 (4.6 mm×50 mm, 1.8 μm), with 0.1% formic acid acetonitrile (A)-0.1% formic acid solution (B) as the mobile phase for gradient elution (0-3 min, 5%-95%A; 3-7 min, 95%A; 7-7.01 min, 95%-5%A; 7.01-8.5 min, 5%A). The flow rate was 0.2 mL·min-1, and the injection volume was 5 μL. In a negative ionization mode, MRM scanning mode was adopted for quantification. Result::The calibration curves of L-shikonin, β, β′-dimethylacrylshikonin, β-acetoxy-isovalerylshikonin, lithospermic acid, caffeic acid, rosmarinic acid showed a good linearity, and the linear ranges of the above six compounds were 10.12-1 012 μg·L-1(r=0.998 1), 10.88-1 088 μg·L-1(r=0.991 2), 10.08-806.4 μg·L-1(r=0.997 6), 20.32-1 016 μg·L-1(r=0.996 6), 10.37-1 037 μg·L-1(r=0.999 6), 10.26-1 026 μg·L-1(r=0.997 8), respectively. The average recoveries of the analysts were 95.8%(RSD 3.2%), 103.5%(RSD 2.3%), 105.3%(RSD 2.1%), 96.1%(RSD 3.3%), 98.9%(RSD 2.7%), 100.8%(RSD 3.4%). The contents of six components in 13 batches of samples showed significant differences. Conclusion::The established method is feasible and simple, and provides a basis for comprehensive quality evaluation of Arnebiae Radix.
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Objective: To investigate the effect of water-soluble components from Salvia miltiorrhiza on liver sinusoid endothelial cell function and angiogenesis. Methods: Different dosages of water-soluble components from S. miltiorrhiza were incubated for 24 h with HHSEC cell line, and the toxicity of HHSEC cells was assayed by CCK-8 method. The proliferation of HHSEC cells was induced by endothelial cell growth supplements (ECGS), with receptor tyrosine kinase inhibitor sorafenib as positive control, cell proliferation was analyzed by EdU DNA cell proliferation kit; Fluorescence probe method was used to assay the intracellular NO level and NOS activity; Immunofluorescence method was performed to observe the expression of CD31; The transgenic zebrafishes were incubated for 24 h with each component. The fluorescence vessels, the number of functional blood vessels, and intersegmental vessel changes were observed; Chicken chorioallantoic membranes were incubated for 24 h with each component, Image Analysis Software was used to analyze the vessel area changes. Results: Compared with control group, the proliferation of HHSEC cells induced by ECGS increased, the level of NO and NOS activity reduced and the expression of CD31 increased; Compared with the model group, salvianolic acid A, salvianolic acid B, salvianolic acid C, and sorafenib inhibited the proliferation of HHSEC cells induced by ECGS, reduced the level of NO, NOS activity, and expression of CD31. The vessel area of chicken chorioallantoic membranes was decreased in sorafenib, salvianolic acid A, salvianolic acid B and caffeic acid. Salvianolic acid C also significantly inhibited the intersegmental vessel changes of transgenic zebrafish. Conclusion: Salvianolic acid A, salvianolic acid B, and salvianolic acid C have the potential effects on function of endothelial cell proliferation and angiogenesis.
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This study was aimed to establish a HPLC method for the content determination of liposoluble components, such as dihydrotanshione I, croptotanshinone, tanshinone I and tanshinone IIA, as well as the content determination of water-soluble components, such as danshensu, protocatechuic aldehyde, rosmarinc acid, salvianolic acid B, and salvianolic acid A in Danshen capsule simultaneously. For liposoluble components, the determination was performed on a Welch ultimate XB-C18 column (250 mm × 4.6 mm, 5 μm) by gradient elution using acetonitrile-water as the mobile phase. The flow rate was 1 mL·min-1. And the detection wavelength was set at 270 nm. For water-soluble components, the determination was performed on a Thermo Syncronis C18 column (250 mm × 4.6 mm, 5μm) by gradient elution using methanol-acetonitrile-0.02%phosphoric acid as the mobile phase. The flow rate was 1 mL·min-1. And the detection wavelength was set at 286 nm. The results showed that there were good linear relationships between components of peak areas and the ranges were 0.472-9.44 μg (r = 0.999 8) for danshensu, 0.352-7.04 μg (r = 0.999 9) for protocatechuic aldehyde, 0.244-4.88 μg (r = 1.000 0) for rosmarinc acid, 2.268-45.36 μg (r = 0.999 9) for salvianolic acid B, 0.168-3.36 μg (r = 0.999 9) for salvianolic acid A, 0.088-1.76 g(r=0.999 9) for dihydrotanshione I, 0.18-3.6μg (r=0.999 9) for croptotanshinone, 0.208-4.16μg (r=0.999 9) for tanshinone I, and 0.17-3.4μg (r=0.999 9) for tanshinone IIA. Average recoveries of the method were between 97.48%and 98.59%. It was concluded that the analysis was stable and reproducible, which can be used as a method for the analysis of Danshen capsule.