壁虎活性组分对HepG2细胞增殖及能量代谢的影响  

Effects of gecko active components on the proliferation and energy metabolism of human hepatocellular carcinoma HepG2 cells

作　　者：黄泽月 段一梦 王兵兵 尹慧萍 王建刚[1] HUANG Ze-yue;DUAN Yi-meng;WANG Bing-bing;YIN Hui-ping;WANG Jian-gang(The Key Laboratory of Pharmacology and Medical Molecular Biology,Medical College,Henan University of Science and Technology,Luoyang 471023,Henan Province,China)

机构地区：[1]河南科技大学医学院药理学与分子生物学实验室

出　　处：《中国临床药理学杂志》2019年第23期3056-3058,3063,共4页The Chinese Journal of Clinical Pharmacology

基　　金：河南省科技重点攻关基金资助项目(142102310031)

摘　　要：目的研究壁虎活性组分(GACs)对人肝癌HepG2细胞增殖及能量代谢的影响。方法将HepG2细胞分为空白组和低、中、高3个剂量实验组。空白组和低、中、高3个浓度实验组分别予以含0,0.10,0.15和0.22 mg·mL-1GACs和2%胎牛血清的培养液进行培养。用噻唑蓝比色法检测GACs对HepG2细胞增殖能力的影响,用分光光度法检测己糖激酶(HK)、丙酮酸激酶(PK)活力及腺苷三磷酸(ATP)的含量,用Western blot法检测磷酸烯醇式丙酮酸羧激酶1(PCK1)、葡萄糖转运蛋白1(GLUT1)、HK2和丙酮酸激酶2(PKM2)的表达。结果GACs作用于HepG2细胞24,48和72 h后,其半抑制浓度分别为0.26,0.22和0.21 mg·mL-1。空白组和低、中、高3个剂量实验组的HK活性分别为(16.71±4.38),(10.96±0.42),(8.59±0.82)和(2.31±1.01)U·g prot-1,PK活性分别为(25.11±0.58),(16.51±0.63),(8.41±1.29)和(8.66±1.24)U·g prot-1,ATP水平分别为(152.54±16.15),(97.52±1.23),(68.49±5.27)和(62.44±11.05)μm·g prot-1,PCK1与GAPDH的灰度值比值分别为(0.14±0.03),(0.16±0.02),(0.54±0.02)和(0.83±0.03),GLUT1与GAPDH的灰度值比值分别为(1.19±0.02),(1.01±0.03),(0.64±0.01)和(0.63±0.02),低、中、高剂量实验组的上述指标和空白组比较,差异均有统计学意义(均P<0.05)。结论GACs可抑制肝癌HepG2细胞能量代谢,其机制与抑制有氧糖酵解,恢复HepG2细胞糖异生有关。Objective To investigate the effects of gecko active components(GACs)on the proliferation and energy metabolism of human hepatocellular carcinoma HepG2 cells.Methods The HepG2 cells were divided into blank and experimental-L,-M,-H groups,which were cultured with 0,0.10,0.15 and 0.22 mg·mL-1 GACs and 2%fetal bovine serum,respectively.Methyl thiazolyl tetrazolium assay was used to observe the change of proliferation ability.The total activities of hexokinase(HK)and pyruvate kinase(PK),and the content of adenosine triphosphate(ATP)were tested by chemical colorimetry.Western blot was applied to observe the expressions of phosphoenolpyruvate carboxy kinase 1(PCK1),glucose transporter 1(GLUT1),HK2,pyruvate kinase 2(PKM2)proteins in HepG2 cells.Results The half maximal inhibitory concentration values of HepG2 cells treated with GACs for 24,48,72 h were 0.26,0.22 and 0.21 mg·mL-1.The activities of HK in blank group and experimental-L,-M,-H groups were(16.71±4.38),(10.96±0.42),(8.59±0.82)and(2.31±1.01)U·g prot-1,the activiteies of PK in above four groups were(25.11±0.58),(16.51±0.63),(8.41±1.29)and(8.66±1.24)U·g prot-1,ATP were(152.54±16.15),(97.52±1.23),(68.49±5.27)and(62.44±11.05)μm·g prot-1,grayseale average ratios of PCK1 and GAPDH were(0.14±0.03),(0.16±0.02),(0.54±0.02)and(0.83±0.03),grayseale average ratios of GLUT1 and GAPDH were(1.19±0.02),(1.01±0.03),(0.64±0.01)and(0.63±0.02),respectively.The differences were statistically significant between experimental-L,-M,-H groups and blank group(all P<0.05).Conclusion GACs can inhibit proliferation and energy metabolism,which may be associated with the inhibition of aerobic glycolysis and the recovery of gluconeogenesis in HepG2 cells.

关 键 词：壁虎活性组分 人肝癌HEPG2细胞 糖异生 有氧糖酵解 

分 类 号：R28[医药卫生—中药学]