title: [Inhibition of mouse hepatocyte apoptosis by anti-caspase-12 small interfering RNA].
abstract: OBJECTIVE
  To study the inhibition of primary mouse hepatocyte apoptosis by small interfering RNA (siRNAs) against caspase-12.
  
  
  METHODS
  The Balb/c mouse primary hepatocytes were isolated in situ with two-step liver perfusion with 0.5 g/L collagenase type IV, and apoptosis were induced with 4 micromol/L thapsigargin (TG). The three kingds of siRNAs targeting different gene sites (130, 214, 521) were synthetized chemically. The single-stranded RNAs were annealed to produce double-stranded siRNAs, then the mouse primary hepatocytes were transfected by oligofectamine package. The inhibition of caspase-12 was analyzed with RT-PCR and Western-blot. The viable hepatocytes following the induction of apoptosis were evaluated with MTT.
  
  
  RESULTS
  All the three kinds of siRNAs could obviously inhibit normal mouse hepatocyte caspase-12 mRNA. The siRNA (214) were more effective than the other two when the concentration was 100 nmol/L. The caspase-12 mRNA expression was inhibited by 52.08%, while that of siRNA (521) was 30.73% (t=4.30, P <0.05). However when the concentration was 200 nmol/L, the inhibitions were similar (88.07%, 86.22% and 89.41% respectively). siRNA (214) could downregulate the expression of apoptotic hepatocytes procaspase-12 by 51.43% ( t=4.30, P <0.01). Contrasted with apoptotic hepatocytes, the cell activity, which was analyzed with MTT, increased by 48.76% (t=2.23, P <0.01).
  
  
  CONCLUSIONS
  siRNAs could effectively downregulate the expression of caspase-12 at mRNA and protein levels and prevent mouse primary hepatocytes from apoptosis.
authors.affiliation: Department of Infectious Diseases, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China.
authors.forename: Hai-fang
authors.initials: HF
authors.lastname: Liu
authors.forename: Qing
authors.initials: Q
authors.lastname: Xie
authors.forename: Shan
authors.initials: S
authors.lastname: Jiang
authors.forename: Guang-ming
authors.initials: GM
authors.lastname: Li
authors.forename: Xia-qiu
authors.initials: XQ
authors.lastname: Zhou
authors.forename: Yi
authors.initials: Y
authors.lastname: Shi
authors.forename: Hong
authors.initials: H
authors.lastname: Yu
authors.forename: You-xin
authors.initials: YX
authors.lastname: Jin
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D053454:Caspase 12 / Q000096:biosynthesis* / Q000235:genetics; D022781:Hepatocytes / Q000166:cytology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D053454:Caspase 12