启膈散乙酸乙酯提取物对食管癌Eca9706细胞增殖和凋亡的影响  

Effect of Qigesan Extracted by Ethyl Acetate on the Proliferation and Apoptosis of Eca9706 Cell in Esophageal Neoplasm

作　　者：尹素改[1] 王慧慧[1] 吴耀松[1] 陈玉龙[1] 

机构地区：[1]河南中医学院科研处分子生物实验中心,河南省郑州市450046

出　　处：《中国全科医学》2015年第14期1663-1666,共4页Chinese General Practice

基　　金：国家自然科学基金面上项目(81173177);河南省高等学校重点科研项目(15A310020)

摘　　要：目的探讨启膈散乙酸乙酯提取物对食管癌Eca9706细胞增殖和凋亡的影响。方法按沙参∶丹参∶茯苓∶川贝母(去心)∶郁金∶砂仁壳∶杵头糠=6∶6∶2∶3∶1∶1∶1的比例配制启膈散,每克药物配10 ml的比例加入95%乙醇溶液,用旋转蒸发仪浓缩药液,浓缩液与乙酸乙酯按体积比2∶1进行萃取,获取启膈散乙酸乙酯提取物。Eca9706细胞与浓度分别为12.5、25.0、50.0、100.0、200.0μg/ml的启膈散提取物共培养,MTT法检测启膈散提取物对Eca9706细胞的抑制率。Eca9706细胞与35%抑制浓度(IC35)、50%抑制浓度(IC50)和70%抑制浓度(IC70)的启膈散提取物共培养,采用流式细胞仪检测启膈散提物对Eca9706细胞的凋亡率。结果不同浓度启膈散提取物对Eca9706细胞的抑制率比较,差异有统计学意义(P<0.05)。启膈散提取物对Eca9706细胞的抑制率与启膈散提取物浓度拟合曲线方程为Y=-1.835×10^(-5)X2+0.006X+0.185,R2=0.996,概率法计算IC35为22.8μg/ml,IC50为81.8μg/ml,IC70为162.2μg/ml。显微镜下观察,随着启膈散提取物浓度的增加,细胞变圆、变小,贴壁细胞数目逐渐减少,细胞间隙增大,细胞碎片增多。IC35、IC50和IC70启膈散提取物对Eca9706细胞的凋亡率比较,差异有统计学意义(P<0.001)。激光共聚焦显微镜下显示,启膈散提取物作用于Eca9706细胞后,部分细胞膜呈绿色,提示早期凋亡;部分细胞膜呈绿色,且细胞核呈红色,细胞体积变小、变圆,形成凋亡小体,提示晚期凋亡细胞。结论启膈散乙酸乙酯提取物可抑制食管癌Eca9706细胞增殖,诱导细胞凋亡。Objective To investigate the effect of qigesan extracted by ethyl acetate on the proliferation and apoptosis of Eca9706 cell in esophageal neoplasm. Methods Qigesan used in the study was confected by the set recipe（ adenophora stricta∶ red sage root∶ poria cocos∶ fritillaria cirrhosa with core removed∶ curcuma aromatic∶ fructus amomi shuck∶ rice bran = 6∶ 6∶ 2∶ 3∶ 1∶ 1∶ 1）. With the proportion of 1 gram of qigesan/ 10 ml ethanol solution,qigesan was mixed with 95% ethanol solution. A rotary evaporator was used to concentrate the mixed liquor. After mixing the concentrated liquor with ethyl acetate as the volume ratio of 2∶ 1,we got the finished qigesan extractive. Eca9706 cells was co - cultured with the extractives of 12. 5,25. 0,50. 0, 100. 0 and 200. 0 μg/ ml concentrations,and MTT method was used to test the inhibition ratios of Eca9706 cells caused by the extractives of different concentrations. Then Eca9706 cells were co - cultured with the extractives of 35% inhibition concentration （IC35）,50% inhibition concentration（IC50）and 70% inhibition concentration（IC70）,and flow cytometry was employed to test the apoptosis rates of Eca9706 caused by the extractives. Results Qigesan extractives of different concentrations induced different（P 〈 0. 05）inhibition ratios of Eca9706 cells. The curve - fitting equation for inhibition ratios of Eca9706 cells and the concentrations of qigesan extractives was Y = - 1. 835 ×10 - 5 X2 ＋ 0. 006X ＋ 0. 185（ R2 = 0. 996）. After calculating by the＆amp;nbsp;equation,we found that the concentrations for IC35,IC50 and IC70 were 22. 8 μg/ ml,81. 8 μg/ ml and 162. 2 μg/ ml respectively. Through a microscope,we observed that,as the concentration of qigesan extractives increasing,Eca9706 cells became rounder and smaller,the amount of adherent cells decreased,the intercellular space expanded and the cell debris increased. IC35,IC50 and IC70 extractives caused significantly different（ P 〈 0. 001 ）apoptosis rates of Eca9706

关 键 词：食管肿瘤 启膈散 细胞增殖 Eca9706细胞 细胞凋亡 

分 类 号：R735.1[医药卫生—肿瘤]