摘要
目的:体外观察药用植物中提取的白黎芦醇与5-氟尿嘧啶联用对人胃癌SGC7901细胞增殖的干预。方法:实验于2003-04/2004-05在重庆医科大学完成。①实验设空白对照组、白藜芦醇治疗组、5-氟尿嘧啶治疗组、药物联合作用组。白藜芦醇(购自美国Sigma公司),配制成11.36mmol/L的储备液,4℃保存备用。5-氟尿嘧啶(南通精华制药厂生产)。人胃癌细胞株SGC7901(重庆医科大学病理生理教研室提供)。②人胃癌癌细胞株SGC7901接种于含体积分数为0.1新生牛血清的RPMI-1640培养液中进行培养,取对数生长期的SGC7901细胞制成5×107L-1的悬液,每孔200μL加入96孔培养板中。③白藜芦醇治疗组按5个不同浓度分别加在96孔板内,每孔加药量20μL,使白黎芦醇的终浓度为28.4,56.8,113.6,227.2,454.4μmol/L。同理5-氟尿嘧啶治疗组的终浓度为48,96.1,192.2,384.4,768.8μmol/L。药物联合作用组分别加入白藜芦醇、5-氟尿嘧啶各100μL,两药合用比例为1∶1,每种单药剂量浓缩1倍。空白对照组则加入等量相同的培养液,不含任何药物。每种剂量3个平行孔。④采用四甲基偶氮唑盐法观察不同浓度白黎芦醇、5-氟尿嘧啶单独或联合应用对人胃癌SGC7901细胞生长的抑制作用,并利用中效原理判定两药合用的效果。结果:①药物抑制率测定结果:随着药物浓度的增加,白黎芦醇、5-氟尿嘧啶单用及联合作用72h时对人胃癌SGC7901细胞的抑制率均相应增加。②白黎芦醇与5-氟尿嘧啶单用、联合作用72h时的回归方程及对SGC7901细胞中效剂量的测定结果:白黎芦醇与5-氟尿嘧啶各自单用时对SGC7901细胞的中效剂量分别为107.08μmol/L和180.06μmol/L;联合用药时为193.74μmol/L,其中白黎芦醇为71.98μmol/L,5-氟尿嘧啶为121.76μmol/L;白黎芦醇单用是合用的1.49倍,5-氟尿嘧啶单用是合用的1.48倍。对于SGC7901细胞,抑制效应大于0.85时合用指数<1,即两药合用时产生协同作用。结论:在体外联合应用白黎芦醇与5-氟尿嘧啶,两种药物很小剂量就能达到各自单用但需很大剂量时才能产生的效果。两药联合应用抑制效应大于0.85时能够产生协同作用。
AIM: To study the intervention of combination of resveratrol and 5-flurouracil (5-FU) on cell proliferation of human SGC7901 carcinoma in vitro.
METHODS: The experiment was performed at Chongqing University of Medical Sciences from April 2003 to May 2004. ①It was designed four groups, including blank control group, resveratrol treated group, 5-FU treated group, combination of resveratrol mad 5-FU group. The resveratrol was purchased from American Sigma Company and made into 11.36 mmol/L reserving liquid and kept at 4℃. 5-FU was produced by Nantong General Pharmaceutical Factory. Human SGC7901 carcinoma cell line was provided by Staff Room of Pathology and Physiology, Chongqing University of Medical Sciences.②Human gastric cancer cell line SGC7901 were grown in RPMI-1640 supplemented with 10% fetal calf serum. Cells in logarithmic phase were made into 5×10^7 L^-1 suspended liquid and seeded into 96-well plates at the 200 μL per well.③Resveratrol treated groups was plused into 96-well plates according to 5 different concentration, and the drug volume that was plused into each well of 96-well plates was 20μL. End concentration of resveratrol was 28.4, 56.8, 113.6, 227.2 and 454.4 μmol/L, so the end concentration of 5-FU was 48, 96.1,192.2, 384.4 and 768.8 μmol/L. The 100μL of each drug was plused in combination of resveratrol and 5- FU group. The rate of combination was 1:1. The dose of each drug was concentrated 1 times. Equal culture liquid that it had nothing of drugs was plused into 3 parallel wells of the blank control group. ④MTT method was used to estimate the inhibitive effect of resveratrol, 5-FU only or combination of resveratrol and 5-FU on the growth of human gastric cancer cell SGC7901 of different concentration, and the median-effect principle was used to analyze union of medication.
RESULTS: ①Result of inhibitive rate: With the increase of drug concentration, the inhibitive rates of resveratrol only, 5-FU only and the combination on human gastric cancer cell SGC7901 increased.②Regression equation of resveratrol only, 5-FU only and the combination as well as result of median-effect dose on human gastric cancer cell SGC7901 at hour 72: The median-effect dose on human gastric cancer cell SGC7901 was 107.08 μmol/L and 180.06 μmol/L with resveratrol only and 5-FU only, respectively; it was 193.74μmol/L with their combination. The resveratrol was 71.98 μmol/L, while 5-FU was 121.76μmol/L. Only resveratrol was 1.49 times of the combination, while only 5-FU was 1.48 times of the combination. For SGC7901 cells, the combination was less than 1 when the inhibitive effect was more than 0.85, which was that there was synergistic action of the combination.
CONCLUSION: A small dose of the combination of resveratrol and 5-FU can have the effect of a large dose of each drug alone in vitro. There is synergistic action when the inhibitive effect of the combination is over 0.85.
出处
《中国临床康复》
CSCD
北大核心
2006年第23期127-129,共3页
Chinese Journal of Clinical Rehabilitation