title: [Total saponins of Chaenomeles speciosa inhibit the degranulation of primary mouse bone marrow-derived mast cells in vitro].
abstract: OBJECTIVE
  To study the effects of total saponins of Chaenomeles speciosa on the degranulation of mouse bone marrow-derived mast cells (BMMCs).
  
  
  METHODS
  Bone marrow cells were isolated from C57BL/6 mice and were cultured in RPMI1640 medium containing 100 mL/L fetal bovine serum, 20 ng/mL IL-3 and 40 ng/mL stem cell factor (SCF). After four-week culture, flow cytometry was used to identify the purity of double-positive (CD117(+)FcEpsilonRIα(+)) mast cells; toluidine blue was used to detect the maturity of mast cells. After total saponins of Chaenomeles speciosa was added into the medium of BMMCs, CCK-8 assay was performed to assess the toxic effects of total saponins of Chaenomeles speciosa on BMMCs; the amount of β-hexosaminidase release was detected by fluorescence spectrophotometry; the content of tryptase in cell supernatants was detected by ELISA.
  
  
  RESULTS
  After cultured for four weeks, the purity of double-positive cells was more than 95%, and the cells presented the features of mature mast cells, with blue nuclear and purple cytoplasm by toluidine blue staining. After BMMCs were cultured in the presence of total saponins of Chaenomeles speciosa (0.01, 0.03, 0.10 mg/mL) for 12 hours, CCK-8 assay indicated that the total saponins did not exert the toxic effects on the BMMCs. Fluorescence spectrophotometry revealed that the release amount and release rate of β-hexosaminidase decreased in the cells treated with the total saponins of Chaenomeles speciosa, DNP-BSA and A23187 compared with the controls, and ELISA assay showed that the amount of tryptase release in the supernatants was reduced as well.
  
  
  CONCLUSIONS
  Total saponins of Chaenomeles speciosa can inhibit the degranulation of primary mouse BMMCs stimulated by different antigens with a clear dose-effect relationship.
authors.affiliation: Medical College, China Three Gorges University, Yichang 443002, China.
authors.forename: Yunli
authors.initials: Y
authors.lastname: Jia
authors.affiliation: Medical College, China Three Gorges University, Yichang 443002, China.
authors.forename: Shigang
authors.initials: S
authors.lastname: Li
authors.affiliation: Medical College, China Three Gorges University, Yichang 443002, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Medical College, China Three Gorges University, Yichang 443002, China.
authors.forename: Lingling
authors.initials: L
authors.lastname: Yu
authors.affiliation: Medical College, China Three Gorges University, Yichang 443002, China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zeng
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects; D015550:Cell Degranulation / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005260:Female; D008407:Mast Cells / Q000187:drug effects* / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D027824:Rosaceae / Q000737:chemistry*; D012503:Saponins / Q000494:pharmacology*
chemical_list: D012503:Saponins