吴茱萸碱对黑色素瘤细胞的耐药逆转作用

黑色素瘤是目前恶性程度最高的肿瘤之一。临床上,常采用维罗非尼（PLX4032）作为晚期患者的治疗药物,但患者很快就出现了耐药。因此,如何克服耐药、提高患者生存率成为急需解决的问题。本文选用中药吴茱萸碱（EVO）与黑色素瘤A375细胞、对PLX4032耐药的A375细胞（A375/R）进行相关研究。采用CCK-8法检测发现,用EVO的细胞非毒性浓度0.5 μmol/L处理A375/R,PLX4032对A375/R细胞的半数抑制浓度（IC50）降低,逆转倍数为3.85。显示EVO能够增强黑色素瘤A375/R细胞对PLX4032的敏感性。后续实验分为对照组、EVO组、PLX组、PLX + EVO组。流式细胞仪检测结果显示,EVO组细胞凋亡率为5.88%,PLX组细胞凋亡率为17.88%,PLX + EVO组细胞凋亡率为30.28%。细胞集落结果证明,PLX4032联合EVO能够抑制A375/R细胞的克隆形成。免疫印迹法结果证明,联合用药能够上调促凋亡蛋白质Bax、胱天蛋白酶3的表达量,下调p-Akt、p-NF-κB-p65及抗凋亡蛋白Bcl-2的蛋白质水平。以上结果均表明,EVO能够有效逆转黑色素瘤A375/R细胞耐药,并且诱导细胞凋亡,抑制细胞增殖。     

Migration,clogging, and carbon source release of nano emulsified vegetable oil in porous media,evaluated by column experiments

To effectively solve the problem of aquifer clogging in the process of in situ bioremediation of groundwater pollution by micron emulsified vegetable oil (Micron EVO), Nano emulsified vegetable oil (Nano EVO) was prepared to replace conventional micron EVO, and three one-dimensional laboratory columns packed with medium or fine sands were conducted to simulate migration, clogging, and carbon source release of EVO in porous media. Column experiment results show that micron and nano EVO resulted in a 20.40% and 3.20% reduction in permeability of medium sand, respectively. Correspondingly, the interception of micron and nano EVO in medium sand were 28.51% and 20.15%, respectively. Obviously, EVO interception is an important reason for permeability loss, and reducing EVO droplet size can effectively alleviate permeability loss in porous media. The COD ratios (dissolved COD/total COD) of micron and nano EVO in medium sand were 87.61% and 61.95%, respectively. The release effect of nano EVO was better than that of micron EVO. Effective longevity of micron and nano EVO were 243.17 d and 98.80 d, respectively. The effect of fine sand media on EVO indicated nano EVO can be used in a finer granular media, and its longevity can also be extended in this media.     

吴茱萸碱对黑色素瘤细胞的耐药逆转作用

黑色素瘤是目前恶性程度最高的肿瘤之一。临床上,常采用维罗非尼（PLX4032）作为晚期患者的治疗药物,但患者很快就出现了耐药。因此,如何克服耐药、提高患者生存率成为急需解决的问题。本文选用中药吴茱萸碱（EVO）与黑色素瘤A375细胞、对PLX4032耐药的A375细胞（A375/R）进行相关研究。采用CCK-8法检测发现,用EVO的细胞非毒性浓度0.5 μmol/L处理A375/R,PLX4032对A375/R细胞的半数抑制浓度（IC50）降低,逆转倍数为3.85。显示EVO能够增强黑色素瘤A375/R细胞对PLX4032的敏感性。后续实验分为对照组、EVO组、PLX组、PLX + EVO组。流式细胞仪检测结果显示,EVO组细胞凋亡率为5.88%,PLX组细胞凋亡率为17.88%,PLX + EVO组细胞凋亡率为30.28%。细胞集落结果证明,PLX4032联合EVO能够抑制A375/R细胞的克隆形成。免疫印迹法结果证明,联合用药能够上调促凋亡蛋白质Bax、胱天蛋白酶3的表达量,下调p-Akt、p-NF-κB-p65及抗凋亡蛋白Bcl-2的蛋白质水平。以上结果均表明,EVO能够有效逆转黑色素瘤A375/R细胞耐药,并且诱导细胞凋亡,抑制细胞增殖。     

Effects of evodiamine and rutaecarpine on the secretion of corticosterone by Zona fasciculata‐reticularis cells in male rats

Evodiamine (EVO) and rutaecarpine (RUT) are two bioactive alkaloid isolated from Chinese herb named Wu–Chu–Yu. Previous studies have shown that EVO and RUT possess thermoregulation, vascular regulation, anti‐allergic, anti‐nociceptive and anti‐inflammatory activities. The mechanisms of EVO and RUT effect on steroidogenesis are not clear. The goal of this study was to characterize the mechanism by which EVO and RUT affect corticosterone production in rat zona fasciculata‐reticularis (ZFR) cells. ZFR cells were isolated from adrenal glands of male rats and incubated with adrenalcorticotropin (ACTH, 10^?9 M), forskolin (an adenylyl cyclase activator, 10^?5 M), 8‐bromo‐adenosine 3′:5′‐cyclic monophosphate (8‐Br‐cAMP, a permeable cAMP analog, 10^?4 M), or steroidogenic precursors including 25‐hydroxycholesterol, pregnenolone, progesterone, and deoxycorticosterone, 10^?5 M each, in the presence or absence of EVO and RUT respectively (0–10^?3 M) at 37°C for 1 h. The concentrations of corticosterone, pregnenolone and progesterone in the media were measured by radioimmunoassay. After administration of ZFR cells with EVO or RUT (10^?4 M) for 60 and 120 min, Western blot analysis was employed to explore the influence of EVO and RUT on the expression of cytochrome P450 side chain cleavage enzyme (P450scc) and steroidogenic acute regulatory protein (StAR). EVO and RUT reduced both basal and ACTH‐, forskolin‐, as well as 8‐Br‐cAMP‐stimulated corticosterone production in rat ZFR cells. The enhanced corticosterone production caused by the administration of four steroidogenic precursors was decreased following EVO or RUT challenge. These results suggest that EVO and RUT inhibit corticosterone production in rat ZFR cells via a mechanism involving: (1) a decreased activity of cAMP‐related pathways; (2) a decreased activity of the steroidogenic enzymes, that is, 3β‐hydroxysteroid dehydrogenase (3β‐HSD) and 11β‐hydroxylase (P450c11), during steroidogenesis; and (3) an inhibition of StAR protein expression. J. Cell. Biochem. 108: 469–475, 2009. © 2009 Wiley‐Liss, Inc.