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左金方对幽门螺旋杆菌感染的人GES-1细胞增殖与凋亡的作用研究

Effect of Zuojin Formula on proliferation and apoptosis of human GES-1 cells infected by Helicobacter pylori
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摘要 目的研究左金方对幽门螺旋杆菌引起人正常胃黏膜上皮(GES-1)细胞增殖与凋亡的影响。方法幽门螺旋杆菌以不同的感染复数(1∶1、50∶1、100∶l、200∶1、300∶1)感染GES-1细胞,于12、24、48 h后,CCK-8检测细胞的增殖活性;以不同质量浓度(0.5、1.0、2.0、4.0μg/m L)左金方作用幽门螺旋杆菌感染的GES-1细胞12、24、48 h后,收集细胞,CCK-8检测细胞的增殖活性,流式细胞仪和Western blotting检测细胞的凋亡率;Hochest染色检测凋亡细胞的形态学改变。结果幽门螺旋杆菌感染GES-1细胞后,感染复数越大,侵染时间越长,抑制细胞生长和促进细胞凋亡的作用越强,幽门螺旋杆菌以100∶1的感染复数作用人GES-1细胞12、24、48 h后,细胞存活率分别降低到(80.57±1.21)%、(70.04±3.21)%、(67.74±2.91)%,细胞的凋亡率分别增加到(23.74±1.71)%、(53.60±1.87)%、(70.67±2.87)%;与1.0μg/m L左金方共培养24 h后,细胞存活率上升到(97.67±1.04)%,细胞凋亡率降低到(31.04±1.02)%,与相应模型组对比,差异显著(P<0.01);Westernblotting结果表明,左金方各处理组细胞Caspase-3蛋白水平表达均明显降低;Hochest荧光染色,从形态学的角度进一步验证了这一结果。结论幽门螺旋杆菌感染人GES-1细胞后,可以抑制细胞增殖,诱导细胞凋亡,左金方可以降低幽门螺旋杆菌引起的细胞损伤,保护GES-1细胞。 Objective To study the effect of Zuojin Formula on proliferation and apoptosis of human normal gastric epithelial cells(GES-1)cells infected by Helicobacter pylori.Methods GES-1 cells were infected by H.pylori at different multiplicity of infection(1∶1,50∶1,100∶1,200∶1,300∶1)for 12,24,48 h;GES-1 cells infected by H.pylori were treated with different concentrations(0.5,1.0,2.0,4.0μg/mL)of Zuojin Formula,and the cells were harvested after 12,24,and 48 h.The proliferation activity of cells was detected by CCK-8,and the apoptosis rate of GES-1 cells was measured by Anexin V-FITC apoptosis detecting kit and flow cytometry.Moreover,the expression of Caspase-3 was detected by Western blot and the morphological changes of apoptotic cells were detected by Hochest staining.Results After infected by H.pylori,the cell viability showed a descending trend while the apoptotic rate showed tendency to ascend,with the increasing of multiplicity of infection and infection time.GES-1 cells were treated with multiplicity of infection of H.pylori at 100∶1 for 12,24,48 h,and the cell viability decreased to(80.57±1.21)%,(70.04±3.21)%,and(67.74±2.91)%,while the apoptotic rate increased to(23.74±1.71)%,(53.60±1.87)%,and(70.67±2.87)%.After co-cultured with 1.0μg/m L Zuojin Formula for 24 h,the cell viability increased to(97.67±1.04)%,and the apoptotic rate decreased to(31.04±1.02)%,and the difference was statistically significant compared with the corresponding model group(P<0.01).The results of Western blotting showed that the protein expression of Caspase-3 of all treatment groups was obviously decreased.Based on the morphological point of view,this result was further verified.Conclusion GES-1 infected by H.pylori NCTC11637 can inhibit cell proliferation and induce apoptosis,while ZuoJinFang could protect GES-1 cells from cell damage.
作者 周红祖 黎莹斯 陈永恒 侯梅轩 余惠旻 ZHOU Hong-zu;LI Ying-si;CHEN Yong-heng;HOU Mei-xuan;YU Hui-min(Shenzhen Chinese Medicine Hospital,Shenzhen 518020,China;School of Medicine,Shenzhen University,Shenzhen 518060,China)
出处 《中草药》 CAS CSCD 北大核心 2019年第4期880-887,共8页 Chinese Traditional and Herbal Drugs
基金 深圳市知识创新基础研究(自由探索)(JCYJ20170818094033689) 深圳大学教学改革项目(JG2017035) 深圳大学聚徒+创研项目(2018延续) 中药特色技术传承人才培训项目(粤中医办函[2016]54号).
关键词 左金方 幽门螺旋杆菌 人正常胃黏膜上皮细胞 细胞增殖 细胞凋亡 Zuojin Formula Helicobacter pylori human normal gastric epithelial cells proliferation apoptosis
作者简介 周红祖,女,副主任中药师,主要从事中药学基础及中药药理、鉴定研究。E-mail:zhouhzu@163.com;通信作者:余惠旻,男,博士,副研究员,主要从事中药及其复方与幽门螺杆菌的研究。E-mail:yuhuimin@szu.edu.cn
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