苦参碱对多发性骨髓瘤细胞凋亡及泛素蛋白酶体通路的影响  

Impact of Matrine on Apoptosis of Multiple Myeloma and Ubiquitin-proteasome Pathway

作　　者：吴迪炯[1] 徐瑾玉[2] 叶宝东[1] 俞庆宏[1] 高雁婷[1] 刘文宾[1] 周郁鸿[1] 

机构地区：[1]浙江中医药大学第一临床医学院 [2]常州市武进中医医院

出　　处：《中医杂志》2016年第10期874-878,共5页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81503296);浙江省自然科学基金(LY14H290001);浙江省中医药优秀青年人才基金计划(2013ZQ012)

摘　　要：目的观察苦参碱对多发性骨髓瘤细胞株凋亡的影响及可能作用机制。方法以不同浓度的苦参碱(0.25、0.5、1.0、2.0 mg/ml)作用于人多发性骨髓瘤细胞株RPMI 8226和U266,另设蛋白酶体抑制剂MG132为阳性对照。采用MTT法测定苦参碱对两种细胞增殖的抑制率,流式细胞仪检测两种细胞的早期凋亡水平,Western-blot法检测两细胞株泛素(Ub)、20S核心颗粒(p Ab)以及NF-κB信号通路相关蛋白(p-NF-κB p65B、IκBα、p-IκBα、IKKα)表达变化。结果在同等浓度苦参碱作用下,两细胞株细胞在48 h的增殖抑制率较本浓度24 h时明显增加(P<0.05或P<0.01)。随着苦参碱浓度的递增,各细胞株的凋亡率均逐渐增加(P<0.05或P<0.01)。随着苦参碱作用浓度的递增,RPMI 8226细胞Ub水平明显上调,同时p Ab呈进行性下降;而U266细胞Ub表达无明显变化,但p Ab水平同样表达下降。苦参碱能明显抑制两细胞株胞核内p-NF-κB p65B以及胞浆内p-IκBα蛋白表达水平,上调RPMI 8226胞浆内IκBα表达,且以浓度为2.0 mg/ml时作用最明显(P<0.05或P<0.01)。结论苦参碱能促进多发性骨髓瘤细胞的凋亡,并且有一定的浓度时间依赖性,其中的关键因素可能是苦参碱具有抑制蛋白酶体活性的作用。Objective To observe the impact of matrine on apoptosis of multiple myeloma and the possible mechanism. Methods Human multiple myeloma cell lines RPMI 8226 and U266 were given matrine in different concentration（ 0. 25,0. 5,1. 0,or 2. 0 mg / ml）. Proteasome inhibitor MG132 was set to be positive control. The inhibition rate of matrine to multiplication of two types of cells was determined adopting methyl thiazolyl tetrazolium（ MTT）method. Early apoptosis level of two types of cells was detected by flow cytometry. Expression changes of ubiquitin（ Ub）,20 S core particle（ p Ab） and NF-κB signaling pathway related proteins（ p-NF-κB p65 B,IκBα,p-IκBα,IKKα） of two cell lines were detected with Western-blot method. Results Under the action of matrine in the same concentration,the inhibitory rate of cell proliferation of two cell lines at the 48 th hour was much higher than that at the 24 th hour（ P 〈 0. 05 or P 〈 0. 01）. With the increasing of matrine concentration,the apoptosis rate of all cell lines increased gradually（ P 〈 0. 05 or P 〈 0. 01）. With the increasing of matrine effective concentration,Ub level in RPMI 8226 cells up-regulated and p Ab decreased progressively. While Ub expression did not change obviously,but p Ab expression decreased in U266 cells. Matrine might inhibit the expressions of p-NF-κB p65 B inside cell nucleus and p-IκBαinside cytoplasm and up-regulate the expression of IκBαinside RPMI 8226 cytoplasm in two cell lines,especially when the concentration was 2. 0 mg / ml（ P 〈 0. 5 or P 〈 0. 01）. Conclusion Matrine might promote apoptosis of multiple myeloma cells with some concentration-time dependence. The critical factor might be that matrine inhibits the activity of proteasome.

关 键 词：多发性骨髓瘤 苦参碱 细胞凋亡 泛素 蛋白酶体 

分 类 号：R285[医药卫生—中药学]