三七提取物对经MNNG转化后的GES-1细胞增殖抑制及促凋亡作用

作者	单位
李军祥	北京中医药大学东方医院消化内科
王志斌	北京中医药大学东方医院消化内科
朱陵群	北京中医药大学东直门医院中医内科学重点实验室
朱福玲	北京中医药大学东直门医院中医内科学重点实验室
崔巍	北京中医药大学东直门医院中医内科学重点实验室

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中文摘要:

目的通过细胞培养方法,研究灌服三七提取物后犬血清对胃癌前病变细胞增殖抑制及促凋亡作用。方法采用永生化的人胃黏膜上皮细胞系GES-1细胞被N-甲基-N-硝基-N-亚硝基胍(MNNG)进一步转化后的细胞(简称MC细胞)作为胃癌前病变细胞的体外研究模型;以三七提取物给彼格犬一次性灌胃,取给药前的血清及给药后不同时点的血清作为实验药物血清。以四甲基固氮唑盐(MTT)法分别检测每个采血时间点上含药血清对MC细胞作用72h后的抑制情况,找出最佳含药血清时点(2h、6h);根据以上结果,以流式细胞仪(flowcytometry)分别检测2h和6h药物血清对MC细胞作用72h后的促凋亡作用。结果2h和6h时点的药物血清对MC细胞的抑制率最高,分别达到45.3%和42.4%,与空白血清组比较差异有显著性(P<0.01);其能明显促进MC细胞的凋亡,与空白血清组比较其凋亡比例增加(P<0.05)。在2h和6h药物血清的作用下MC细胞在G0/G1期细胞比例明显下降(P<0.05);而G2/M期细胞显著增加(P<0.05);但S期并没有一致性的变化。结论灌服三七提取物后2h、6h药物血清对MC细胞的增殖抑制作用最强;均对MC细胞有显著的促凋亡作用,且能降低MC细胞在G0/G1期的比例及增加G2/M期的比例,这可能是其抑制MC细胞增殖及促凋亡的原因之一。

英文摘要:

ObjectiveTo study the effect drug contained canine serum, prepared by gastric perfusion with Sanchi extract （SE）, in inhibiting proliferation and promoting apoptosis of cultured precancerous gastric cells by cell culture. MethodsThe precancerous model cells （MC） used in the experiment were prepared through transforming eternalized human gastric mucosa epithelial cells GES-1 by N-methyl-N’-nitro-N-nitroso-guanidine （MNNG）. After once gastric perfusion of SE extract to dogs, the canine serum gotten before and at different time points after medication was used for test. The inhibitory effect of the drug serum obtained at different time points on MC after acting for 72 hrs was detected by 3-（4,5）-dimethy thioazol-2-yl-2,5-diphenyl-tetrazoliumbromide （MTT） method to find the optimal time point for drug serum preparation, that were 2 hrs and 6 hrs after medication. Then the cell apoptosis promoting effect after acting for 72 hrs of the drug serum obtained at the optimal time points was determined by flow cytometry. ResultsThe drug serum obtained at 2-hr and 6-hr after medication showed the highest inhibitive effect on MC cells, reaching 45.3% and 42.4% respectively, as compared with the effect of blank serum, the difference was significant （<0.01）. They could evidently promote the MC cell apoptosis, the apoptosis rate also showed significant difference to that of the blank serum （<0.05）. Under their action, the proportion of MC cells in G₀/G₁ phase was obviously decreased （<0.05） while that in the G₂/M phase significantly increased （PWTBZ<0.05）. However, the change of cells in S phase was not uniform. ConclusionThe drug contained canine serum gotten 2 hr and 6 hr after SE feeding shows the optimal MC proliferation inhibitive effect and significant apoptosis promoting effect. Besides, it could significantly decrease the proportion of MC cells in G₀/G₁ phase and significantly increase that in G₂/M phase, this effect might be one of the mechanisms of ES in inhibiting MC cell proliferation and promoting its apoptosis.

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