摘要
目的:探讨广藿香醇(PA)诱导MV4-11细胞凋亡,并探讨其可能存在的相关活性。方法:采用四甲基偶氮唑盐比色(MTT)法检测PA对人结、直肠癌细胞HCT116,人肝癌细胞Hep G-2,人肺癌细胞A549,人急性淋巴髓单核细胞白血病细胞MV4-11,人皮肤黑色素瘤细胞A375,小鼠乳腺癌细胞4T1,人单核细胞型淋巴瘤细胞THP-1,人正常胚肾293A细胞的增殖抑制作用;采用Hoechst 33258染色法观察细胞凋亡的形态学变化;通过Annexin V-FITC/碘化丙啶(PI)双染法并应用流式细胞术检测细胞凋亡;免疫印迹法(Western blot)检测磷酸化M2型丙酮酸激酶(p-PKM2),核转录因子-κB(NF-κB),半胱氨酸天冬氨酸蛋白酶3(Caspase-3)凋亡蛋白的表达。结果:PA能抑制人HCT116,Hep G-2,A549,MV4-11,A375,4T1,THP-1,293A细胞的增殖,半抑制浓度(IC50)分别为0.26,0.32,0.28,0.09,0.37,0.23,0.24,0.33 mmol·L-1;作用于MV4-11细胞24 h后可使细胞核皱缩,染色质凝聚,形成明显的凋亡小体。0.1,0.2,0.5 mmol·L-1PA能诱导MV4-11细胞凋亡,凋亡率分别为7.9%,13.6%,22.0%,与溶剂组比较,结果具有统计学差异(P<0.05);Western blot检测显示,PA使细胞NF-κB,p-PKM2,Caspase-3蛋白表达发生明显改变(P<0.05)。结论:PA能抑制人白血病细胞MV4-11细胞的增殖,并诱导其发生凋亡,作用机制可能与NF-κB,p-PKM2,Caspase-3蛋白量的改变有关。
Objective： To study the effect of patchouli alcohol（ PA） on inducing apoptosis of MV4-11 cells and preliminarily investigate its potential mechanism. Method： The 3-（ 4,5-dimethyl-2 thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide（ MTT） assay was used to detect the inhibitory effect of PA on human colorectal cancer cell line HCT116,human hepatocellular carcinoma cell line Hep G-2,human lung epithelial cell line A549,human leukemia cell line MV4-11,human skin melanoma cell line A375,mice breast cancer cell line 4T1,human mononuclear cell type lymphoma cell line THP-1,and human normal embryonic kidney cell line 293 A. The morphology changes of apoptosis were observed by Hoechst 33258 staining method. Apoptosis was detected by Annexin V-FITC / Propidium Iodide staining method using flow cytometry. Western blot was used to detect the protein expressions of NF-κB,p-PKM2 and Cystine aspartic acid protease-3（ Caspase-3）. Result： PA could efficiently inhibit the proliferation of HCT116,Hep G-2,human A549,MV4-11,A375,4T1,THP-1,and 293 A,with IC50 values of 0. 26,0. 32,0. 28,0. 09,0. 37,0. 23,0. 24,and 0. 33 mmol·L- 1,respectively. After MV4-11 cells were treated for 24 h,nuclear shrinkage,chromatin condensation and obvious apoptosis were observed.The 0. 1,0. 2,and 0. 5 mmol·L- 1PA could induce apoptosis of MV4-11 cells,with the apoptosis rates of 7. 9%,13. 6%,22. 0% respectively,with statistically significant difference compared with control group（ P 〈 0. 05）.Western blot results showed that PA could result in changes of NF-κB,p-PKM2,and Caspase-3 protein expressions（ P〈 0. 05）. Conclusion： PA can inhibit the proliferation of human leukemic cell line MV4-11,and induce apoptosis. Its mechanism may be associated with the changes of NF-κB, p-PKM2, and Caspase-3 protein expressions.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第6期99-103,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81403149)
四川省科技厅项目(2011BAI13B05)
