title: Regulation of apoptosis through bcl-2/bax proteins expression and DNA damage by Zanthoxylum alatum.
abstract: BACKGROUND
  Many of the major chemotherapeutic agents are secondary metabolites found in nature. Zanthoxylum alatum Roxb. (Rutaceae) is traditionally used in the treatment of various diseases.
  
  
  OBJECTIVE
  The present study evaluates the apoptotic activity of methanol extract of Z. alatum (MEZA) on Ehrlich ascites tumor (EAT) in Swiss albino mice.
  
  
  METHODS
  The presence of flavonoids in MEZA was standardized by HPLC. The in vitro cytotoxicity of MEZA was measured by the MTT assay. The in vivo antitumor activity of MEZA (100 and 200 mg/kg b.w., i.p. for 9 days) was also evaluated. On the 10th day, EAT tumor volume, cell viability, and hematological parameters were assayed. Apoptotic morphology was determined by acredine orange/ethedium bromide using fluorescence microscopy. Apoptosis percentage was measured by flow cytometric analysis using annexine-V-FITC. Also, DNA damage and bcl-2/bax were estimated by UV-method and western blot, respectively.
  
  
  CONCLUSIONS
  HPLC analysis revealed presence of three flavonoids, rutin, myricetin, and quercetin. MEZA showed satisfactory cytotoxicity in MTT assay (IC50 = 111.50 µg/ml). The extract significantly (p < 0.01) changed the tumor volume, viable, non-viable cell count, and hematological parameters towards the normal. Apoptotic activity of MEZA was confirmed by acridine orange/ethidium bromide staining, annexin-V-FITC staining, DNA fragmentation, and Bcl-2/Bax ratio.
  
  
  CONCLUSIONS
  The study showed that MEZA has antitumor activity which may be due to the presence of flavonoids in the extract.
authors.affiliation: a Department of Pharmaceutical Technology , Jadavpur University , Kolkata , West Bengal , India and.
authors.forename: Indrajit
authors.initials: I
authors.lastname: Karmakar
authors.affiliation: a Department of Pharmaceutical Technology , Jadavpur University , Kolkata , West Bengal , India and.
authors.forename: Sagnik
authors.initials: S
authors.lastname: Haldar
authors.affiliation: a Department of Pharmaceutical Technology , Jadavpur University , Kolkata , West Bengal , India and.
authors.forename: Mainak
authors.initials: M
authors.lastname: Chakraborty
authors.affiliation: b Indian Institute of Chemical Biology , Kolkata , West Bengal , India.
authors.forename: Keya
authors.initials: K
authors.lastname: Chaudhury
authors.affiliation: a Department of Pharmaceutical Technology , Jadavpur University , Kolkata , West Bengal , India and.
authors.forename: Saikat
authors.initials: S
authors.lastname: Dewanjee
authors.affiliation: a Department of Pharmaceutical Technology , Jadavpur University , Kolkata , West Bengal , India and.
authors.forename: Pallab Kanti
authors.initials: PK
authors.lastname: Haldar
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D002286:Carcinoma, Ehrlich Tumor / Q000188:drug therapy / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation; D008297:Male; D051379:Mice; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D018515:Plant Leaves; D032106:Zanthoxylum; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis*
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D010936:Plant Extracts; D051028:bcl-2-Associated X Protein
keywords: Cytotoxicity; Ehrlich ascites tumor; flavonoids; western blot