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).Conclusion 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.