机构地区:[1]湖南中医药大学针灸推拿学院经穴与脏腑相关重点研究室,长沙410007 [2]深圳市中医院推拿科,深圳518033
出 处:《针刺研究》2011年第3期157-163,192,共8页Acupuncture Research
基 金:国家自然科学基金(No.30772707);教育部博士点科研基金(20070541003)
摘 要:目的:探讨艾灸血浆对胃黏膜细胞凋亡的影响,揭示艾灸对胃黏膜细胞的保护作用及其信号转导机制。方法:24名健康人随机等分为艾灸穴位组和艾灸非穴位组,分别艾灸中脘、关元和足三里穴及非穴位对照点各10 d。将人体胃黏膜上皮(GES-1)细胞分为空白组、模型组、艾灸穴位血浆组、艾灸非穴位血浆组,采用含8%乙醇的培养液造成GES-1细胞损伤模型,分别加以预先提取的灸前人体血浆、艾灸穴位和非穴位血浆。流式细胞技术检测细胞凋亡率;Western-blot法检测细胞内热休克蛋白70(HSP70)、第二个线粒体来源的胱氨酸酶激活物(Smac)、凋亡诱导因子(AIF)的表达;细胞免疫化学法检测半胱氨酸天冬酸蛋白酶-3、9(Caspase-3、9)的表达。结果:模型组与空白组比较,细胞凋亡率明显增加,HSP70、Smac、AIF、Caspase-3、Caspase-9表达均上调(均P<0.01);艾灸穴位血浆组与模型组比较,凋亡率明显下降(P<0.01),HSP70表达进一步提高(P<0.01),Smac、AIF、Caspase-3、Caspase-9表达下调(均P<0.01);艾灸非穴位血浆组与模型组比较,凋亡率、Smac表达下降,Caspase-3、Caspase-9表达也下调(均P<0.01),但高于艾灸穴位血浆组(P<0.05,P<0.01)。结论:艾灸中脘、关元和足三里穴位后提取的人体血浆能抑制GES-1细胞凋亡,其信号转导通路可能是:促进细胞内HSP70合成,通过线粒体凋亡通路,抑制Smac、AIF表达,同时阻断其与Caspase-9的结合,减少Caspase-3含量,抑制细胞凋亡。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 rain, 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 Oaspase (Smac) and AIF proteins of GES-1 cells were assayed by western blotting, and the immunoactivity of cysteinyl aspirate-specific proteinase-3 and 9 (Oaspase-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 Oaspase-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 immunoac
关 键 词:艾灸血浆 人胃黏膜上皮细胞 凋亡 热休克蛋白70 胱氨酸酶激活物 凋亡诱导因子 半胱氨酸天冬酸蛋白酶-3 9
分 类 号:R245.81[医药卫生—针灸推拿学]
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