摘要
目的通过体外实验研究甘草提取物(Gc34)对人肝癌细胞Hep G-2侵袭、增殖生物学行为的影响及其可能的分子机制。方法将人Hep G-2细胞分为对照组、Gc34组。常规培养细胞。对照组用酒精干预,使其终浓度为0.5%;Gc34组用Gc34干预,使其终浓度为12.5、25.0、50.0 mg/L。细胞侵袭实验测定Hep G-2细胞侵袭力。四甲基偶氮唑蓝(MTT)法检测Hep G-2细胞的增殖活性。酶联免疫吸附实验(ELISA)测定细胞培养上清基质金属蛋白酶(MMP)2、9的含量。比色法测定细胞培养上清还原性谷胱甘肽/谷胱甘肽(GSH/GSSG)、超氧化物岐化酶(SOD)、丙二醛(MDA)的含量。结果 HE染色光学显微镜下Hep G-2细胞核浆比例增大,核深染、大小、形态和染色不一,核分裂像增多并可见病理性核分裂像:巨核、双核、多核。24 h及以上的同时间,Gc34 25.0、50.0 mg/L组Hep G-2细胞增殖活性显著低于对照组(P均<0.01)。Gc34组Hep G-2细胞侵袭数、MMP-2、MMP-9、GSH/GSSG、SOD分别为45±7、(40.3±6.2)mg/L、(42.7±5.6)mg/L、4.2±0.8、(67.2±7.9)U/(mg·L),均显著低于对照组(P均<0.01)。Gc34组Hep G-2细胞培养上清MDA含量为(45.4±8.5)mmol/g,显著高于对照组(P<0.01)。Hep G-2细胞侵袭力与MMP-2、MMP-9正相关(r分别为0.65、0.72,P均<0.01)。Hep G-2细胞增殖活性与GSH/GSSG、SOD正相关(r分别为0.55、0.64,P均<0.01),与MDA负相关(r=-0.58,P<0.01)。结论甘草提取物(Gc34)下调MMP-2、MMP-9表达,抑制Hep G-2细胞侵袭能力;下调GSH/GSSG、SOD表达,上调MDA表达,抑制Hep G-2细胞增殖活性。
Objective To investigate the influence of glycyrrhiza（ Gc34） on the invasion and proliferation of Hep G-2 cell and its mechanism. Methods Hep G-2 cells were divided into control group and Gc34 group. Hep G-2 cells were treated with 12. 5,25. 0,50. 0 mg / L Gc34 and 0. 5% alcohol. Cell invasion assay was used to detect the invasion of Hep G-2 cell.The proliferation of Hep G-2 cell was detected by methyl thiazolyl tetrazolium（ MTT）. Matrix metalloproteinase 2,9,GSH /GSSG,MDA and SOD of Hep G-2 cell supernatant were measured. Results The nucleus of Hep G-2 cell under light microscope was abnormal with an increase in the ratio of nucleus to cytoplasm and nucleus fission,nucleus anachromasis,meganucleus,binucleus,polynucleus. Inhibition of Gc34 on the proliferation of Hep G-2 cell was time-dependent and concentration-dependent（ P〈0. 01）. The invasive ability,content of MMP-2,9,GSH / GSSG,SOD of Hep G-2 cell supernatant in Gc34 group was 45 ± 7,（ 40. 3 ± 6. 2） mg / L,（ 42. 7 ± 5. 6） mg / L,4. 2 ± 0. 8,（ 67. 2 ± 7. 9） U /（ mg·L）,respectively,which were significantly lower than those in the control group（ P〈0. 01）. The content of MDA of Hep G-2 cell supernatant in Gc34 group was（ 45. 4 ± 8. 5） mmol / g which was predominantly higher than that in control group（ P〈0. 01）. The invasion positively correlated with MMP-2,MMP-9（ r = 0. 65,0. 72,P〈0. 01）. The proliferation positively correlated with GSH / GSSG,SOD（ r = 0. 55,0. 64,P〈0. 01）,negatively correlated with MDA（ r =- 0. 58,P〈0. 01）. Conclusion Gc34 can inhibit the invasion of Hep G-2 cell by down-regulating MMP-2,9 and attenuate the proliferation by down-regulating GSH / GSSG,SOD and up-regulating MDA.
出处
《中华全科医学》
2016年第8期1249-1251,1318,共4页
Chinese Journal of General Practice
基金
国家自然科学基金青年科学基金(81201672)
浙江省义乌市人才引进立项项目(2012-R-04)
关键词
甘草
HEPG-2细胞
侵袭
增殖
基质金属蛋白酶
Glycyrrhiza
Hep G-2 cell line
Invasion
Proliferation
Matrix metalloproteinase
