益气健脾法对人胃癌MGC803细胞增殖及凋亡的影响

Effects of benefiting qi and strengthening spleen method on proliferation and apoptosis of human gastric cancer MGC803 cells

目的:探讨益气健脾法对人胃癌MGC803细胞增殖及凋亡的影响。方法:20只SPF级雄性大鼠随机分为对照组(n=10)、益气健脾组(n=10)。对照组每日灌胃给予生理盐水1ml/100g,益气健脾组按照每日灌胃给予四君子汤1ml/100g,1次/d,共7d,第7天灌胃给药1.5h后腹主动脉取血分离出血清;采用含15%胎牛血清、15%对照组血清以及15%益气健脾组血清的DMEM培养基分别培养MGC803细胞后,应用CCK-8试剂分别在加药处理24、48及72h时检测细胞的增殖能力;加药处理72h后采用流式细胞仪检测分析各组细胞凋亡率的改变;同时采用Real-time PCR的方法检测各组细胞中Bax-2和Caspase3 mRNA表达水平的改变。结果:与空白对照组及15%对照血清组相比较,15%益气健脾血清组细胞的增殖能力在48h及72h均明显下降(P<0.05);加药72h后,细胞的凋亡率明显增加[(16.38±0.84)%vs(8.32±2.73)%,vs(8.07±2.46)%,P<0.01],细胞中凋亡相关基因Bax和Caspase3 mRNA表达水平显著上调(P<0.05)。结论:益气健脾法能够有效抑制胃癌MGC803细胞的增殖能力并促进其凋亡,这可能是其治疗胃癌的潜在机制。

Objective: To explore the effects of benefiting qi and strengthening spleen method on proliferation and apoptosis of human gastric cancer MGC803 cells. Methods: 20 SPF male rats were randomly divided into control group( n =10) and the benefiting qi and strengthening spleen group( n =10).Control group were treated with 1 ml/100 g daily of physiological saline and benefiting qi and strengthening spleen group were treated with isodose Sijunzi decoction,once everyday for a week.On the seventh day we drew blood from the abdominal aorta and serum were isolated 1.5 h after drug treatment.MGC803 cells were cultured in DMEM medium containing 15% FBS serum,15% control serum group serum and 15% benefiting qi and strengthening spleen group serum,respectively.Proliferation abilities of MGC803 cells 24 h,48 h and 72 h after drug treatment were tested by CCK-8 assay.And apoptosis rate,Bax and Caspase3 mRNA expression level of the cells were examined by FCM and Real-time PCR 72 h after drug treatment,respectively. Results: Compared with control group and 15% control serum group,proliferation abilities of 15% benefiting qi and strengthening spleen group were decreased 48 h and 72 h after drug treatment( P <0.05).The apoptosis ratio of 15% benefiting qi and strengthening spleen group were significantly increased compared with control group and 15% control serum group[(16.38±0.84)% vs (8.32±2.73)%,vs (8.07±2.46)%, P <0.01].Apoptosis-related genes Bax and Caspase3 mRNA level were all significantly increased( P <0.05). Conclusion: The therapy for benefiting qi and strengthening spleen can inhibit proliferation and promote apoptosis of human gastric cancer MGC803 cells,and it may be a potential mechanism of benefiting qi and strengthening spleen in the treatment of gastric cancer.