头顶一颗珠正丁醇萃取物对人肝癌HepG2细胞的抑制作用及其对MMP-9、NF-κB p65的表达研究

Inhibitory effect of n-butanol extracts from Trillium Tschonoskii Maxim on human hepatoma HepG2 cells and its expression of MMP-9 and NF-κB p65

目的探究头顶一颗珠提取物对人肝癌HepG2细胞抑制作用及其对基质金属蛋白酶-9(matrix metallo proteinases-9,MMP-9)、核转录因子-κB p65(nuclear factorκB p65,NF-κB p65)的表达研究。方法多浓度头顶一颗珠正丁醇萃取物(0、6、12、24、48、96)μmol/L分别作用于人肝癌细胞株(hepatocarcinoma G2,HepG2)24小时、48小时、72小时,使用细胞增殖和毒性检测试剂盒(cell counting kit-8,CCK8)法,比较各组肝癌HepG2细胞的生长抑制率。多浓度头顶一颗珠正丁醇萃取物(0、3、6、12、24)μmol/L分别作用于HepG2细胞48小时后,用实时荧光定量PCR(real-timequantitative PCR,qPCR)检测MMP-9、NF-κB p65的表达水平。结果多浓度头顶一颗珠正丁醇萃取物,与对照组比较,HepG2细胞的生长增殖均受到明显抑制(P<0.01),且呈时间和浓度依赖性;MMP-9、NF-κB p65表达水平明显降低(P<0.01),药物浓度越高,MMP-9 mRNA、NF-κB p65 mRNA表达水平越低,MMP-9、NF-κB p65 mRNA表达水平呈正相关性。结论多浓度头顶一颗珠正丁醇萃取物能抑制人肝癌HepG2细胞增殖,且呈时间和浓度依赖性,可能通过抑制MMP-9 mRNA、NF-κB p65 mRNA的表达,从而达到抗肝癌作用。

Objective To investigate the inhibitory effect of a pearl extract from Trillium Tschonoskii Maxim on hepatocarcinoma G2(HepG2)cells and its expression of matrix metallo proteinases-9(MMP-9)and nuclear factor kappa bp65(NF-κB p65).Methods Multi concentration n-butanol extracts from Trillium Tschonoskii Maxim at a concentration of 0,6,12,24,48 and 96(μmol/L)acted on HepG2 for 24 hours,48 hours and 72 hours respectively,then cell proliferation and toxicity detection Kit(CCK8)was used to compare the growth inhibition rate of HCC HepG2 cells in each group.Multi concentration n-butanol extracts from Trillium Tschonoskii Maxim at a concentration of 0,3,6,12 and 24(μmol/L)acted on HepG2 cells for 48 hours respectively,then the expression levels of MMP-9 and NF-κB p65 were detected by real-time quantitative PCR(qPCR)method.Results When it comes to multi concentration n-butanol extracts from Trillium Tschonoskii Maxim,compared with the control group,the growth and proliferation of HepG2 cells were significantly inhibited(P<0.01),and it is time and concentration dependent,and the expression levels of MMP-9 and NF-κB p65 were significantly reduced(P<0.01).The higher the drug concentration,the lower the mRNA expression levels of MMP-9 and NF-κB p65,and the positive correlation between the mRNA expression levels of MMP-9 and NF-κB p65.Conclusion Multi concentration n-butanol extracts from Trillium Tschonoskii Maxim can inhibit the proliferation of human hepatoma HepG2 cells,which is time and concentration dependent.It is possible to inhibit the mRNA expression of MMP-9 and NF-κB p65,so as to have anti-hepatocellular carcinoma effect.