ABSTRACT
Objective To investigate the effects of crude extract of Capparis spinosa L. fruit alka-loids (CSFA) on the maturation of murine bone marrow-derived dendritic cells (DCs). Methods CSFA was prepared and the contents were determined by high performance liquid chromatography. DCs were trea-ted with different doses (1, 2, 3 mg/ml) of CSFA. The viability of DCs, the expression of surface mole-cules and the ability of phagocytosis were detected by flow cytometry. The secretion of cytokines was meas-ured by ELISA. Western blot assay was performed to analyze the activation of key molecules in mitogen-acti-vated protein kinases ( MAPK) and nuclear factor-kappa B ( NF-κB) signaling pathways. Results The re-sults showed that CSFA alone had no significant influence on the expression of surface molecules and cyto-kines in DCs. However, it significantly decreased the expression of CD40, CD80, CD86 and MHC Ⅱ as well as the secretion of IL-12p40 and TNF-αthat were induced by lipopolysaccharides (LPS), but increased IL-10 secretion and the ability of phagocytosis after treating DCs with both CSFA and LPS. Further, the phosphorylation of p38, extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) and the nuclear translocation of NF-κBp65 induced by LPS were inhibited by CSFA. Conclusion CSFA could inhibit the maturation of DCs and the secretion of pro-inflammatory cytokines induced by LPS while in-creasing the secretion of the anti-inflammatory cytokine IL-10 and the ability of phagocytosis, which might in-volve MAPK and NF-κB signaling pathways. This study suggests that CSFA could be used as a potential im-munosuppressant.
ABSTRACT
AIM: To select proper hydrolysis time to determine Ginkgo biloba flavonoids content. METHODS: Sample dissolved in 25mL MeOH-25%∶HCl(4∶1) mixture and was heated to hydrolyze for various time, then diluted. Flavonoids'content was determined by HPLC. The conditions were developed by Shimpack CLC-ODS(150mm?6mm) column using 0.4% phosphoric acid solution-Methanol(50∶50), the detective wavelength was at 360nm with quercetin、kaempferol and isorhamnetin as standards, temperature being 30℃. RESULTS: Contents of flavonoids of Ginkgo biloba leaves,Ginkgo biloba crude extracts and Ginkgo biloba refined extracts were highest in the hydrolysis time of 2.0h. Ginkgo biloba flavonoids can be completely hydrolyzed in 2.0h. CONCLUSION: The hydrolysis time of 2.0h is best.