title: Cytotoxic effects of four Caryophyllaceae species extracts on macrophage cell lines.
abstract: BACKGROUND
  Saponins have been reported to possess antitumor properties, to inhibit angiogenesis and to induce tumor apoptosis.
  
  
  OBJECTIVE
  To test the possible cytotoxic effect of crude extracts from four Caryophyllaceae species including Gypsophila paniculata L., Gypsophila trichotoma Wend., Saponaria officinalis L., and Dianthus sylvestris Wulffen on cultured monocyte/macrophage cell lines.
  
  
  METHODS
  After acid hydrolysis of the methanol-aqueous extracts, two representative prosaponins of the Caryophyllaceae, gypsogenin 3-O-glucuronide and quillaic acid 3-O-glucuronide were purified using solid-phase extraction (SPE), then identified by ultra-performance liquid chromatography-electrospray/mass spectrometry (UPLC-ESI/MS). Cytotoxic activity of the crude extracts at concentrations ranging from 0.1 to 200 µg/ml was evaluated on rat alveolar macrophage NR8383 and human monocytic THP-1 cell lines. Apoptosis was determined by measuring caspase-3 activity.
  
  
  RESULTS
  Quantitative analysis by reversed-phase high-performance liquid chromatography (RP-HPLC) revealed a high content of gypsogenin 3-O-glucuronide in Gypsophila species roots (0.52-1.13% dry weight). At a concentration ≥10 µg/ml of crude extracts, a significant reduction of NR8383 and THP-1 cell lines viability was evidenced using the Trypan blue exclusion test. D. sylvestris extract exhibited the highest toxicity against THP-1 cells. Caspase-3 activation was evidenced after 4 and 24 h incubation of macrophages with 100 µg/ml of S. officinalis and G. trichotoma extracts, indicating apoptosis induction.
  
  
  CONCLUSIONS
  Crude extracts from the assayed species revealed cytotoxic effects toward macrophage cell lines. In Gypsophila species, gypsogenin 3-O-glucuronide derivatives could be responsible for the observed cytotoxicity. Therefore, crude extract of Caryophyllaceae is worth investigating for the potential development of agents against cancer cells.
authors.affiliation: Department of Pharmacognosy, Faculty of Pharmacy, Medical University-Sofia , Sofia , Bulgaria .
authors.forename: R
authors.initials: R
authors.lastname: Gevrenova
authors.forename: O
authors.initials: O
authors.lastname: Joubert
authors.forename: Tsv
authors.initials: T
authors.lastname: Mandova
authors.forename: M
authors.initials: M
authors.lastname: Zaiou
authors.forename: Y
authors.initials: Y
authors.lastname: Chapleur
authors.forename: M
authors.initials: M
authors.lastname: Henry
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D029748:Caryophyllaceae / Q000737:chemistry*; D053148:Caspase 3 / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D020719:Glucuronides / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D008264:Macrophages / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D000432:Methanol / Q000737:chemistry; D009000:Monocytes / Q000187:drug effects; D009828:Oleanolic Acid / Q000031:analogs & derivatives / Q000032:analysis / Q000302:isolation & purification; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity*; D018517:Plant Roots / Q000737:chemistry; D051381:Rats; D012503:Saponins / Q000032:analysis / Q000302:isolation & purification
chemical_list: D020719:Glucuronides; D010936:Plant Extracts; D012503:Saponins; C062873:gypsogenin 3-O-glucuronide; C005564:quillaic acid; D009828:Oleanolic Acid; D053148:Caspase 3; D000432:Methanol
keywords: Apoptosis; Dianthus sylvestris Wulffen; Gypsophila paniculata L.; Gypsophila trichotoma Wend.; Saponaria officinalis L.; caspase-3; gypsogenin 3-O-glucuronide