四君子汤对脾虚证胃癌MGC803细胞周期凋亡及Caspase-3表达的影响

Effects of Sijunzi Decoction on the cell cycle apoptosis and the expression of Caspase-3 in gastric cancer MGC803 of spleen deficiency syndrome

目的:探讨四君子汤对脾虚证胃癌MGC803细胞周期凋亡及Caspase-3表达。方法:采用倒置显微镜观察人胃癌MGC803细胞形态学变化,透射电镜观察染色情况,采用流式细胞术检测细胞周期及凋亡,采用免疫组化法检测Caspase-3蛋白表达情况。结果:倒置显微镜下5umol/L四君子汤组作用48 h后胃癌细胞数目减少,细胞分裂不明显,体积明显缩小,可发现核固缩现象以及圆形体积变小的凋亡小体形成;透射电镜下5umol/L四君子汤处理后细胞膜微绒毛减少,胞质有所浓缩且细胞体积变小,核染色聚集与核膜,电子密度高,可见明显的凋亡小体;流式细胞仪检测结果显示,四君子汤可诱导MGC803发生凋亡,细胞周期检测显示G0/G1期比值明显增加,而在S其与G2/M期则降低,故在G0/G1期发生阻滞;5umol/L四君子汤组Caspase-3表达显著高于0.1%DMSO组,差异有统计学意义。结论:四君子汤对脾虚证胃癌MGC803具有明显促凋亡作用,细胞周期阻滞在G0/G1期,且通过增加Caspase-3表达而促进其凋亡。

Objective： To investigate Sijunzi decoction Caspase-3 expression and MGC803 gastric cell cycle and apoptosis in spleen deficiency. Methods： MGC803 inverted microscope morphological changes in human gastric cancer cells, transmission electron microscopy staining; using flow cytometry cell cycle and apoptosis; detection of Caspase-3 protein expression using immunohistochemistry. Results： The inverted microscope 5umol / L Sijunzi decoction group for 48h to reduce the number of gastric cancer cells , cell division is not obvious , the volume was significantly reduced, the phenomenon can be found karyopyknosis and smaller size of the circular formation of apoptotic bodies; TEM 5umol / L after treatment Sijunzi decoction membrane microvilli , cytoplasmic somewhat concentrated and smaller cell size , nuclear chromatin aggregation and nuclear membrane , high electron density , showed obvious apoptotic bodies; flow cytometry results Sijunzi decoction MGC803 can induce apoptosis , cell cycle analysis showed that the ratio of G0/G1 phase increased significantly, while the S with the G2 / M phase was reduced , so arrest occurred in the G0/G1 phase; 5umol / L Sijunzi decoction group Caspase-3 expression was significantly higher than 0. 1% DMSO group , and the difference was significant. Conclusion： Sijunzi decoction gastric MGC803 for spleen deficiency has obvious effect on apoptosis and cell cycle arrest in G0/G1 phase and by increasing the expression of Caspase-3 induce apoptosis.