健脾中药胃肠安与5-FU相互作用的中效原理评价
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摘要
目的:采用中效原理探讨健脾复方胃肠安(Weichang′an,WCA)联合5-氟尿嘧啶(5-fluorouracil,5-FU)的相互作用,并观察体外对胃癌细胞MKN45的增殖、凋亡等生物学行为的影响。方法:本实验选取人胃癌细胞株MKN45为实验对象,用不同浓度的WCA、5-FU单药和两药11浓度配比联合作用于胃癌细胞(24 h,48 h,72 h),Cell Counting Kit-8(CCK-8)法检测药物对胃癌细胞抑制作用。根据中效原理分析两药联合作用的效应(Fa)与合用指数(CI)的关系,判定药物间的相互作用。以药物单用及合用的1/2倍中效浓度作用于MKN45细胞,流式细胞凋亡检测法评价药物对细胞周期、凋亡的影响,Hoechst 33258荧光染色观察细胞凋亡形态。结果:两种药物单独应用或联合应用均对胃癌细胞有明显的抑制作用,并且呈剂量效应依赖关系。由中效原理计算可知,两药联合应用抑制胃癌细胞增殖作用增强,存在协同效应。细胞周期检测提示WCA与5-FU均阻滞细胞周期于S期,两药联用时更加显著;流式细胞凋亡检测与Hoechst染色均发现单药组细胞凋亡现象较对照组明显,而联合组细胞凋亡现象更加明显。结论:健脾中药WCA与5-FU联合应用对胃癌MKN45细胞产生协同效应。
Objective:To discuss the interaction between invigorating spleen Chinese medicine and 5-FU using the median effect principle and observe the effect of in vitro on proliferation,apoptosis and other biological behaviors of gastric cancer cell MKN45.Methods:In this experiment,human gastric cancer cell line MKN45 was selected as the experimental object,and different concentrations of WCA,5-FU single drug and 1■1 concentration ratio of the two drugs were combined to act on gastric cancer cells(24 h,48 h,72 h).Cell Counting Kit-8(CCK-8) method was used to detect the relationship between the inhibition of drugs on gastric cancer cells.According to the median effect principle,the relationship between the effect(Fa) of the combined action of the two drugs and the combination index(CI) was analyzed to determine the interaction between the drugs.The MKN45 cells were treated with 1/2 times the median concentration of the drugs used alone and in combination.The effects of drugs on cell cycle and apoptosis were detected by flow cytometry.The morphology of apoptosis was observed by Hoechst 33258 fluorescence staining.Results:Both drugs alone or in combination had obvious inhibitory effect on gastric cancer cells and show a dose-effect dependent relationship.From the calculation of the medium efficiency principle,the combined application of the two drugs enhances the inhibition of gastric cancer cell proliferation and had a synergistic effect.Cell cycle test showed that WCA and 5-FU both blocked cell cycle in S phase,and the combination of WCA and 5-FU was more significant.Flow cytometry and Hoechst staining showed that apoptosis in the single drug group was more obvious than that in the control group,while apoptosis in the combined group was more obvious.Conclusion:Combined application of invigorating spleen Chinese medicine and 5-FU Jianpi compound WCA and 5-FU has a synergistic effect on MKN45 cells of gastric cancer.
Objective:To discuss the interaction between invigorating spleen Chinese medicine and 5-FU using the median effect principle and observe the effect of in vitro on proliferation,apoptosis and other biological behaviors of gastric cancer cell MKN45.Methods:In this experiment,human gastric cancer cell line MKN45 was selected as the experimental object,and different concentrations of WCA,5-FU single drug and 1■1 concentration ratio of the two drugs were combined to act on gastric cancer cells(24 h,48 h,72 h).Cell Counting Kit-8(CCK-8) method was used to detect the relationship between the inhibition of drugs on gastric cancer cells.According to the median effect principle,the relationship between the effect(Fa) of the combined action of the two drugs and the combination index(CI) was analyzed to determine the interaction between the drugs.The MKN45 cells were treated with 1/2 times the median concentration of the drugs used alone and in combination.The effects of drugs on cell cycle and apoptosis were detected by flow cytometry.The morphology of apoptosis was observed by Hoechst 33258 fluorescence staining.Results:Both drugs alone or in combination had obvious inhibitory effect on gastric cancer cells and show a dose-effect dependent relationship.From the calculation of the medium efficiency principle,the combined application of the two drugs enhances the inhibition of gastric cancer cell proliferation and had a synergistic effect.Cell cycle test showed that WCA and 5-FU both blocked cell cycle in S phase,and the combination of WCA and 5-FU was more significant.Flow cytometry and Hoechst staining showed that apoptosis in the single drug group was more obvious than that in the control group,while apoptosis in the combined group was more obvious.Conclusion:Combined application of invigorating spleen Chinese medicine and 5-FU Jianpi compound WCA and 5-FU has a synergistic effect on MKN45 cells of gastric cancer.
引文
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[19]韦尉元,吴琨,王晓通,等.5-Fu联合人参黄芪复方对人胃癌MGC-803细胞生物学行为的影响[J].中国中西医结合杂志,2012,32(12):1647-1651.
[20]郑秋惠,陈卫国,王法林,等.健脾益肾中药对晚期胃癌化疗患者的辅助疗效观察[J].四川中医,2012,30(3):73-74.
[21]YAMAGUCHI K,NAKAGAWA S,YABUSAKI H,et al.Combination chemotherapy with 5-fluorouracil,cisplatin and paclitaxel for pretreated patients with advanced gastric cancer[J].Anticancer research,2007,27(5):3535-3539.
[22]CHOU T C.Drug combination studies and their synergy quantification using the Chou-Talalay method[J].Cancer research,2010,70(2):440-446.
[23]邱佳信,唐莱娣,左建平,等.中药的反突变作用研究[J].上海中医药杂志,1985,19(9):46-49.
[24]邱佳信,杨金坤,唐莱娣,等.晚期恶性肿瘤的异病同治——健脾法在消化道恶性肿瘤中的应用[J].上海中医药杂志,1984,18(1):2-4.
[25]赵海磊,赵爱光,尤圣富,等.健脾中药复方胃肠安对人胃癌裸小鼠原位移植瘤生长和转移的影响[J].中西医结合学报,2005,3(5):48-51.
[26]赵爱光,杨金坤,赵海磊.健脾中药诱导人胃癌细胞裸小鼠移植瘤细胞凋亡的实验研究[J].世界华人消化杂志,2000,8(7):737-740.
[27]苑伟,陈彬,赵爱光,等.健脾复方对人胃癌细胞株MKN- 45裸小鼠皮下移植瘤模型非特异性免疫功能的影响[J].中华中医药杂志,2013,28(9):2766-2770.
[2]ADAM J,BASS V T,SHMULEVICH I Y,et al.Comprehensive molecular characterization of gastric adenocarcinoma[J].Nature,2014,513(7517):202-209.
[3]许晶,侯丽,武苗,等.新加良附方联合5-Fu对裸鼠胃癌移植瘤Bax/Bcl-2表达的影响[J].世界中西医结合杂志,2013,8(7):675-678.
[4]黄挺,杨雪飞,李勇,等.健脾理气方对人胃癌移植瘤小鼠5-Fu代谢过程的影响[J].中华中医药学刊,2013,31(4):871-874.
[5]吴晓勇,赵林涛,宋延平,等.加味良附丸联合5-Fu对裸鼠胃癌移植瘤NF-ΚBp65表达的影响[J].陕西中医药大学学报,2017,40(1):76-80.
[6]李朝燕,赵爱光.中医药治疗晚期胃癌临床对照试验疗效的文献分析[J].中医药学报,2011,39(5):46-49.
[7]赵爱光,李朝燕,孙姗姗,等.健脾为基础的中药辨证治疗对老年胃癌生存期影响的同期对照研究[J].中国肿瘤,2010,19(10):651-658.
[8]符玉章.中西医结合疗法运用于胃癌术后50例[J].河南中医,2016,36(4):682-683.
[9]ZHAO L,ZHAO AG,ZHAO G,et al.Survival benefit of traditional chinese herbal medicine(a herbal formula for invigorating spleen) in gastric cancer patients with peritoneal metastasis[J].Evid Based Complement Alternat Med,eCAM,2014:625493.
[10]XU Y,ZHAO A G,LI Z Y,et al.Survival benefit of traditional Chinese herbal medicine(a herbal formula for invigorating spleen) for patients with advanced gastric cancer[J].Integrative cancer therapies,2013,12(5):414-422.
[11]朱晓虹,赵爱光,李宏伟,等.基于健脾为基础的辨证治疗方案对ⅢC期胃癌根治术后患者无病生存期的影响[J].中国肿瘤,2016,25(7):569-574.
[12]沈克平,王海永,胡兵,等.胃肠安对人胃癌SGC-7901细胞TGF-β1诱导上皮间质转化作用[J].中国实验方剂学杂志,2011,17(23):132-135.
[13]赵爱光,杨金坤,尤圣富,等.中药胃肠安诱导裸鼠胃癌移植瘤细胞凋亡的途径及基因调控的研究[J].中西医结合学报,2007,5(3):287-297.
[14]CHOU T C,TALALAY P.Quantitative analysis of dose-effect relationships:the combined effects of multiple drugs or enzyme inhibitors[J].Adv Enzyme Regul,1984,22:27-55.
[15]CHOU T C.Theoretical basis,experimental design,and computerized simulation of synergism and antagonism in drug combination studies[J].Pharmacological reviews,2006,58(3):621-681.
[16]郭佳栋,冯变玲,张雪梅,等.胃癌患者5-氟尿嘧啶用药安全性Meta分析[J].中国药业,2015,24(2):27-32.
[17]周阿高,李琰,洪声,等.中医药联合化疗治疗胃癌疗效的Meta分析[J].世界中西医结合杂志,2010,5(5):376-378.
[18]于庆生,经文善,宋加奎,等.丹参及其联合5-Fu对人胃癌SGC-7901细胞凋亡的影响[J].世界中医药,2016,11(7):1315-1319.
[19]韦尉元,吴琨,王晓通,等.5-Fu联合人参黄芪复方对人胃癌MGC-803细胞生物学行为的影响[J].中国中西医结合杂志,2012,32(12):1647-1651.
[20]郑秋惠,陈卫国,王法林,等.健脾益肾中药对晚期胃癌化疗患者的辅助疗效观察[J].四川中医,2012,30(3):73-74.
[21]YAMAGUCHI K,NAKAGAWA S,YABUSAKI H,et al.Combination chemotherapy with 5-fluorouracil,cisplatin and paclitaxel for pretreated patients with advanced gastric cancer[J].Anticancer research,2007,27(5):3535-3539.
[22]CHOU T C.Drug combination studies and their synergy quantification using the Chou-Talalay method[J].Cancer research,2010,70(2):440-446.
[23]邱佳信,唐莱娣,左建平,等.中药的反突变作用研究[J].上海中医药杂志,1985,19(9):46-49.
[24]邱佳信,杨金坤,唐莱娣,等.晚期恶性肿瘤的异病同治——健脾法在消化道恶性肿瘤中的应用[J].上海中医药杂志,1984,18(1):2-4.
[25]赵海磊,赵爱光,尤圣富,等.健脾中药复方胃肠安对人胃癌裸小鼠原位移植瘤生长和转移的影响[J].中西医结合学报,2005,3(5):48-51.
[26]赵爱光,杨金坤,赵海磊.健脾中药诱导人胃癌细胞裸小鼠移植瘤细胞凋亡的实验研究[J].世界华人消化杂志,2000,8(7):737-740.
[27]苑伟,陈彬,赵爱光,等.健脾复方对人胃癌细胞株MKN- 45裸小鼠皮下移植瘤模型非特异性免疫功能的影响[J].中华中医药杂志,2013,28(9):2766-2770.