三七提取物对GES-1细胞及MNNG转化后GES-1细胞增殖的抑制作用

Inhibiting effects of Panax notoginseng extracts on proliferation of GES-1 cells and MNNG-transformed GES-1 cells

目的 :通过细胞培养方法 ,由研究灌服三七提取物后的犬血清对胃癌前细胞的抑制作用入手 ,以期找出三七入血后的最佳作用时间点 (段 )。方法 :采用永生化的人胃黏膜上皮细胞系GES 1细胞以及N 甲基 N 硝基 N 亚硝基胍 (N methyl N’ nitro N nitroso guanidine ,MNNG)进一步转化GES 1细胞后获得的细胞 (以下简称MC细胞 )作为胃癌前病变细胞的体外研究模型 ;以三七水提物给彼格犬一次性灌胃 ,取给药前的血清、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 [3 (4,5 dimethylthiazol 2 yl) 2 ,5 diphenyltetrazoliumbromide ,MTT]法分别检测每个采血时间点上不同浓度 (8%、4 %、2 % )含药血清对GES 1及MC细胞作用 72h后的抑制情况 ,并根据结果找出抑制作用最佳的药物血清时间点。结果 :2h和6h含药血清刺激下的抑制率最高 ,两个时间点含 8%药物血清的培养基对两种细胞的抑制作用均十分显著 ,其中 2h 8%GES 1组抑制率为 70 .8% (P <0 .0 1) ,2h 8%MC组抑制率为 4 5 .3% (P <0 .0 1) ;6h 8%GES 1组抑制率为 88.5 % (P <0 .0 1) ,6h 8%MC组抑制率为 4 2 .4 % (P <0 .0 1)。结论 :灌服三七提取物后 2h、6h取的药物犬血清对GES 1及MC细胞的增殖抑制作用最强。

Objective: Through cell cultivation, we studied the inhibiting effects of the serum of the dog fed with Panax notoginseng extracts on precancerous gastric cells, trying to find the best time points or periods when the extracts’ function was the strongest after administration of the extracts to the dog. Methods: The experiments adopted eternalized human gastric mucosa epithelium GES-1 cells and MC cells gained from GES-1 cells transformed by N-methyl-N’-nitro-N-nitroso-guanidine (MNNG) as the model of precancerous lesions for study in vitro. We took the serum of a dog before and at different points of time after feeding the dog with Panax notoginseng extracts for experiment. By means of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, we examined the inhibiting effects of the serum after culturing the GES-1 and MC cells for 72 hours with different concentration (8%, 4%, 2%) of medicated serum obtained from the dog at different points of time, so as to find that, at which points of time the medicated serum obtained, it would be the most effective. Results: The results showed that the GES-1 and MC cells inhibition rates of medicated serum from the points of 2-hour and 6-hour were the highest, and the culture medium containing 8% of medicated serum from these two points had prominent inhibiting effects on both kinds of cells. The GES-1 cells inhibition rate in culture medium containing 8% of medicated serum from the point of 2-hour was 70.8% (P<0.01) and that of the MC cells was 45.3% (P<0.01). The GES-1 cells inhibition rate in culture medium containing 8% of medicated serum from the point of 6-hour was 88.5% (P<0.01) and that of the MC cells was 42.4% (P<0.01). Conclusion: The points of time with the strongest inhibiting effects are 2 hours and 6 hours after being fed with Panax notoginsengextracts. At these two points, the serum is most effective in inhibiting the proliferation of GES-1 and MC cells.