title: [Effect of plasma of healthy subjects undergoing moxibustion on ethanol-injured human gastric epithelial GES-1 cells in vitro and the involved mitochondrial apoptosis pathway].
abstract: OBJECTIVE
  To observe the effect of plasma derived from healthy volunteers undergoing moxibustion (moxibustion plasma) on alchol-injured human gastric epithelial GES-1 cells in vitro, and expression of heat shock protein 70 (HSP 70, cell apoptosis inhibitory protein), apoptosis inducing factor (AIF), Smac (a mitochondrial protein), and Caspase 3 and Caspase 9 (the latter 3 proteins are also involved in cell apoptosis) in order to study its mechanisms underlying protecting gastric mucous membrane.
  
  
  METHODS
  Twenty-four healthy volunteer subjects (half men and half women) were randomized into acupoint-moximustion (A-M) [Zhongwan(CV 12), Guanyuan (CV 4) and Zusanli (ST 36)] group and non-acupoint-moxibustion (NA-M, 3 cun right to CV 12 and CV 4.1 cun medial to ST 36 ) group (n = 12/group). Moxibustion was applied to the above-mentioned 3 acupoints and non-acupoints for 30 min, once daily for 10 days. Venous blood of the subjects was collected before and after moxibustion. The cultured GES-1 cells were divided into: control group. ethanol-injury group (model), A-M plasma group (A-M-P, plasma got from volunteers undergoing A-M), and NA-M plasma group (NA-M-P,plasma got from volunteers accepting NA-M). The GES-1 cells of the latter 3 groups were treated with 8% ethanol for duplicating cell injury model. Apoptosis was detected by flowcytometry. Expression of HSP 70, second mitochondria-derived activator of Caspase (Smac) and AIF proteins of GES-1 cells were assayed by western blotting, and the immunoactivity of cysteinyl aspirate-specific proteinase-3 and 9 (Caspase-3, 9) was detected by immunocytochemistry.
  
  
  RESULTS
  In comparison with the control group, the apoptosis rate, the expression of HSP 70, Smac and AIF proteins, and the immunoactivity of Caspase-3 and Caspase-9 of the model group were increased significantly (P < 0.01). Compared with the model group, the apoptosis rate of GES-1 cells, the expression of Smac and AIF proteins, and the immunoactivity of Caspase-3 and Caspase-9 in the A-M-P group, the apoptosis rate, the expression of Smac and the immunoactivity of Caspase-3 and Caspase-9 in the NA-M-P group were all down-regulated considerably (P < 0.05, P < 0.01). In comparison with the model group, HSP 70 expression of the A-M-P group was up-regulated significantly (P < 0.01). The apoptosis rate of GES-1 cells, the expression levels of Smac, AIF, Caspase-3 and Caspase-9 were significantly lower in the A-M-P group than in the NA-M-P group (P < 0.05, P < 0.01), while the expression of HSP 70 was apparently higher in the A-M-P group than in the NA-M-P group (P < 0.01).
  
  
  CONCLUSIONS
  Plasma derived from the subjects undergoing moxibustion of Zusanli (ST 36), Zhongwan (CV 12) and Guanyuan (CV 4) can inhibit apoptosis of GES-1 cells in vitro, which is closely related to its effects in up-regulating intracellular HSP 70 expression and down-regulating mitochondrial apoptosis protein expression of AIF. Smac, Caspase-3 and Caspase-9.
authors.affiliation: Lab for Studying Meridian-Zangfu-organ Correlation, College of Acu-moxibustion and Massage, Hunan University of Chinese Medcine, Changsha 410007, China. hjbywzywz@126.com
authors.forename: Jinbiao
authors.initials: J
authors.lastname: Hong
authors.forename: Shou-Xiang
authors.initials: SX
authors.lastname: Yi
authors.forename: Yun
authors.initials: Y
authors.lastname: Huang
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Lin
authors.forename: Yan
authors.initials: Y
authors.lastname: Du
authors.forename: Hong
authors.initials: H
authors.lastname: Peng
authors.forename: Yan
authors.initials: Y
authors.lastname: Peng
mesh_terms: D000293:Adolescent; D000328:Adult; D017209:Apoptosis; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053453:Caspase 9 / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D000431:Ethanol / Q000633:toxicity; D005260:Female; D005753:Gastric Mucosa / Q000166:cytology* / Q000187:drug effects / Q000293:injuries / Q000378:metabolism*; D005786:Gene Expression Regulation; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D008297:Male; D008928:Mitochondria / Q000378:metabolism*; D009071:Moxibustion; D010949:Plasma / Q000737:chemistry*; D055815:Young Adult
chemical_list: D018840:HSP70 Heat-Shock Proteins; D000431:Ethanol; D053148:Caspase 3; D053453:Caspase 9