姜半夏乙醇提取物对人胃腺癌SGC7901细胞内pH值的影响  

Effects of ethanol extract of Rhizome Pinelliae Preparata on intracellular pH value of human gastric adenocarcinoma cells

作　　者：张慈安[1] 武峰[1] 毛竹君[1] 魏振[1] 李勇进[1] 魏品康[1] 

机构地区：[1]第二军医大学长征医院中医科,上海200003

出　　处：《中西医结合学报》2011年第8期894-900,共7页Journal of Chinese Integrative Medicine

摘　　要：目的:观察姜半夏乙醇提取物对人胃腺癌SGC7901细胞内pH值的影响。方法:不同浓度姜半夏乙醇提取物(终浓度分别为1、0.5、0.25、0.125mg/mL)处理SGC7901细胞后,通过甲基噻唑基四唑(3-(4,5-di methylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide,MTT)法检测细胞增殖状况,使用2,7-双(2-羧基乙基)-5(6)-羧基荧光素四(丙酰甲酯)(2,7-bis-(2-carboxyethyl)-5-carboxyfluorescein-acetoxymethyl,BCECF-AM)荧光探针检测细胞内pH值,pH计测定胞外培养基的pH值;采用荧光定量聚合酶链反应技术检测药物干预细胞72h后细胞钠氢交换蛋白1(Na+/H+exchangerisoform1,NHE1)和空泡质子转运ATP酶(vacuolar-H+-ATPase,V-ATPase)基因的表达水平。结果:不同浓度姜半夏乙醇提取物均能不同程度地抑制人胃癌SGC7901细胞的增殖,降低细胞内pH值,提高细胞外培养基pH值。0.5mg/mL姜半夏乙醇提取物可下调SGC7901细胞V-ATPase和NHE1mRNA的表达。结论:姜半夏乙醇提取物具有降低SGC7901细胞内pH值的作用,其机制可能与抑制细胞V-ATPase和NHE1mRNA表达有关。Objective：To observe the effects of ethanol extract of Rhizome Pinelliae Preparata on the intracellular pH value of human gastric cancer SGC7901 cells.Methods：After coculturing SGC7901 cells with ethanol extract of Rhizome Pinelliae Preparata（1,0.5,0.25 and 0.125 mg/mL）,cell viability was evaluated by chromatometry with 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT）staining.Intracellular pH value of SGC7901 cells was measured in the monolayer by using the pH-sensitive fluorescent probe 2,7-bis-（2-carboxyethyl）-5-carboxyfluorescein-acetoxymethyl ester.The extracellular pH value of culture medium was measured by a pH211 Calibration Check Microprocessor pH Meter.Half-inhibitory concentration（IC50）of ethanol extract culture to SGC7901 cells was decided by the MTT method and expressions of vacuolar-H＋-ATPase（V-ATPase）and Na＋/H＋ exchanger isoform 1（NHE1）mRNAs were examined by the method of fluorescence quantitative-polymerase chain reaction after 72 h of drug treatment.Results：Ethanol extract of Rhizome Pinelliae Preparata at different concentrations significantly inhibited the proliferation of SGC7901 cells,lowered the intracellular pH values and heightened the extracellular pH values.The IC50 of 72 h culture was 0.5 mg/mL and it inhibited the expressions of V-ATPase and NHE1 mRNAs.Conclusion：Ethanol extract of Rhizome Pinelliae Preparata can lower down the intracellular pH value of SGC7901 cells.The mechanism may be related to inhibiting the expressions of V-ATPase and NHE1 mRNAs.

关 键 词：中草药 半夏 胃肿瘤 细胞内PH值 钠氢反向转运物 空泡质子转运ATP酶 细胞系 肿瘤 

分 类 号：R285.5[医药卫生—中药学]