title: [Active ingredients of Plastrum Testudinis inhibit epidermal stem cell apoptosis in serum-deprived culture].
abstract: OBJECTIVE
  To investigate the effects of active ingredients of Plastrum Testudinis (PT) on serum deprivation-induced apoptosis of epidermal stem cells (ESCs).
  
  
  METHODS
  ESCs were isolated from the back skin of fetal Sprague-Dawley rats with 2 weeks of gestational age and were divided into normal group (10% fetal bovine serum), control group (serum-deprived culture) and groups treated with serum deprivation plus active ingredients of PT, including ethyl acetate extract (2B), stearic acid ethyl ester (S6), tetradecanoic acid sterol ester (S8) and (+)-4-cholesten-3-one (S9). The vitality of ESCs after 24, 48 and 72 h of culture was measured with MTT method; apoptotic ESCs double-stained with Annexin V-FITC and propidium iodine were detected by flow cytometry (FCM); Bcl-2 and caspase-3 expressions were measured by Western blotting.
  
  
  RESULTS
  MTT results indicated that the vitality of ESCs in the active ingredients of PT groups at 48 h was increased compared with the control group and 2B had better effects than the others. FCM results indicated that 2B had the most significant anti-apoptotic effect compared with the control as well as S6, S8 and S9. Western blot results indicated that 2B, S6, S8 and S9 up-regulated the expression of Bcl-2 protein and down-regulated the expression of caspase-3 protein compared with the control.
  
  
  CONCLUSIONS
  Ethyl acetate extract of Plastrum Testudinis inhibits epidermal stem cell apoptosis in serum-deprived culture by regulating the expressions of Bcl-2 and caspase-3 proteins and has a stronger anti-apoptotic effect than its constituents S6, S8 and S9.
authors.affiliation: Teaching and Research Section of Anatomy, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Chen
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.forename: Chun
authors.initials: C
authors.lastname: Li
authors.forename: Jia-hui
authors.initials: JH
authors.lastname: Cao
authors.forename: Yi-ling
authors.initials: YL
authors.lastname: Wu
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Jian-hong
authors.initials: JH
authors.lastname: Zhou
authors.forename: Dong-feng
authors.initials: DF
authors.lastname: Chen
authors.forename: Yi-wei
authors.initials: YW
authors.lastname: Li
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects*; D008297:Male; D008419:Materia Medica / Q000494:pharmacology*; D008516:Medicine, Chinese Traditional; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*
chemical_list: D016895:Culture Media, Serum-Free; D008419:Materia Medica; D019253:Proto-Oncogene Proteins c-bcl-2; C505410:Casp3 protein, rat; D053148:Caspase 3