裂果薯皂苷Ⅰ通过干预谷氨酰胺代谢影响人肝癌BEL-7404细胞增殖的研究  

Saponins from Schizocapsa Plantaginea Hance I affects the proliferation of human hepatocellular carcinoma BEL-7404cellsby interfering with glutamine metabolism

作　　者：何能婷 秦燕萍 李昊 吕美娴 莫宇雪 李锦华 植丽敏 左琪琪 王金妮[1] 梁梓樱 郭心怡 王燕雪 梁钢[1] HE Neng-ting;QIN Yan-ping;LI Hao;LU Mei-xian;MO Yu-xue;LI Jin-hua;ZHI Li-min;ZUO Qi-qi;WANG Jin-ni;LIANG Zi-ying;GUO Xin-yi;WANG Yan-xue;LIANG Gang(College of Pharmacy,Guangxi Medical University,Nanning 530000,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学药学院,广西壮族自治区南宁530000

出　　处：《中国临床药理学杂志》2023年第9期1232-1236,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81460620、81960737)。

摘　　要：目的探究裂果薯皂苷Ⅰ(SSPHⅠ)对BEL-7404细胞的增殖及谷氨酰胺代谢的影响,并分析其可能的作用机制。方法体外培养BEL-7404细胞,分别给予不同浓度的SSPHⅠ(0、1、2、3、4、5、6、7和8μmol·L^(-1))处理24、48和72 h,用噻唑蓝法检测SSPHⅠ对BEL-7404细胞的存活率的影响。将BEL-7404细胞分为空白组(不做任何处理)和低、中、高剂量实验组(分别给予2、4和8μmol·L^(-1)SSPHⅠ处理)。用酶联免疫吸附实验法检测细胞内谷氨酰胺(Gln)、谷氨酸(Glu)和谷胱甘肽(GSH)的含量,用流式细胞术检测细胞内活性氧(ROS)水平,用蛋白质印迹法检测谷氨酰胺酶(GLS1)及谷氨酸脱氢酶(GLUD1)蛋白的表达水平。结果SSPHⅠ对肝癌BEL-7404细胞的增殖有抑制作用。SSPHⅠ作用于BEL-7404细胞24、48、72 h后的半抑制浓度分别为3.37、2.60和2.70μmol·L^(-1)。低、中、高剂量实验组和空白组的细胞克隆形成率分别为(76.53±0.93)%、(18.60±0.49)%、(8.00±1.23)%和(80.20±0.82)%,Gln含量分别为(1728.89±37.47)、(1526.11±20.79)、(1434.44±38.69)和(1812.22±37.47)μmol·L^(-1),Glu含量分别为(51.97±0.06)、(47.95±0.61)、(45.91±0.59)和(55.24±0.54)mg·L^(-1),GSH含量分别为(110.13±1.84)、(102.91±1.58)、(69.40±1.01)和(150.05±1.33)μmol·L^(-1),ROS含量分别为(31.53±2.32)%、(34.90±2.06)%、(52.12±3.99)%和(22.67±2.15)%,GLS1蛋白相对表达水平分别为0.90±0.13、0.61±0.08、0.43±0.10和1.28±0.16,GLUD1蛋白相对表达水平分别为1.28±0.09、0.92±0.22、0.65±0.03和1.34±0.24。高剂量实验组的上述指标与空白组比较,差异均有统计学意义(均P<0.01)。结论SSPHⅠ可以抑制BEL-7404细胞的增殖,其机制可能与抑制谷氨酰胺代谢有关。Objective To investigate the effect of Saponins from Schizocapsa Plantaginea Hance I(SSPH I)on the proliferation and glutamine metabolism of BEL-7404 cells,and analyze its possible mechanism.Methods BEL-7404 cells were cultured in vitro and treated with different concentrations of SSPH I(0,1,2,3,4,5,6,7,8μmol·L^(-1))for 24,48 and 72 h,respectively.Methyl thiazolium tetrazolium assay was used to observe the viability of the BEL-7404 cells.BEL-7404 cells were divided into blank group(without any treatment)and experimental-L,-M,-H groups(treated with 2,4,8μmol·L^(-1)SSPH I,respectively).The levels of intracellular glutamine(Gln)and its related metabolites glutamate(Glu)and glutathione(GSH)were measured by enzyme-linked immunosorbent assays,and the levels of intracellular reactive oxygen species(ROS)in BEL-7404 cells were measured by flow cytometry.The protein levels of glutaminase(GLS1)and glutamate dehydrogenase(GLUD1)were measured by Western Blot method.Results SSPH I could inhibit the proliferation of BEL-7404 cells,the half maximal inhibitory concentration(ICso)of SSPH I on BEL-7404 cells after 24,48 and 72 h were 3.37,2.60,2.70μmol·L^(-1),respectively.The cell clone formation rates of experimental-L,-M,-H groups and blank group were(76.53±0.93)%,(18.60±0.49)%,(8.00±1.23)%and(80.20±0.82)%;Gln contents in the 4 groups were(1728.89±37.47),(1526.11±20.79),(1434.44±38.69)and(1812.22±37.47)μmol·L-;Glu contents in the 4 groups were((51.97±0.06),(47.95±0.61),(45.91±0.59)and(55.24±0.54)mg·L^(-1);GSH contents in the 4 groups were(110.13±1.84),(102.91±1.58),(69.40±1.01)and(150.05±1.33)μmol·L^(-1);R0S contents in the 4 groups were(31.53±2.32)%,(34.90±2.06)%,(52.12±3.99)%and(22.67±2.15)%;the relative expression levels of GLS1 protein in the 4 groups were 0.90±0.13,0.61±0.08,0.43±0.10 and 1.28±0.16;the relative expression levels of CLUD1 protein in the 4 groups were 1.28±0.09,0.92±0.22,0.65±0.03 and 1.34±0.24,respectively.Comparing with the blank group,the above indexes in the experimen

关 键 词：裂果薯皂苷Ⅰ 肝癌 增殖 谷氨酰胺代谢 

分 类 号：R28[医药卫生—中药学]