蛇床子素对Y1小鼠肾上腺皮质瘤细胞皮质功能的影响  

Effect of Osthole on Adrenocortical Function in Y1 Mouse Adrenocortical Tumor Cells

作　　者：潘志强[1] 梁龙龙[1] 方肇勤[1] 刘小美[1] 卢文丽[1] 张园园[1] 

机构地区：[1]上海中医药大学基础医学院,上海201203

出　　处：《中国中西医结合杂志》2016年第5期574-579,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81473562);上海市进一步加快中医药事业发展三年行动计划(2014-2016年)(No.ZY3-CCCX-3-3010)

摘　　要：目的研究蛇床子素(Osthole,Ost)对肾上腺皮质功能的影响。方法以Y1小鼠肾上腺皮质瘤细胞为实验对象,以含10%、1%和0.1%血清培养液培养细胞,分别采用1、10、25、50、100、200μmol/L Ost处理Y1细胞24、48 h,以0.1%二甲基亚砜(DMSO)为阴性对照组,腺苷酸环化酶激活剂(Bu)2cAMP为阳性对照组,倒置显微镜下观察细胞形态变化,采用ELISA法检测细胞分泌液皮质酮含量,RT-qPCR法检测类固醇急性调节蛋白(Star)、胆固醇侧链裂解酶(Cyp11a1)、21α-羟化酶(Cyp21a1)、3β-羟基类固醇脱氢酶(Hsd3b2)、11β-羟化酶1(Cyp11b1)、11β-羟化酶2(Cyp11b2)、17α-羟化酶/17,20-碳链裂解酶(Cyp17a1)及17β-羟基类固醇脱氢酶3(Hsd17b3)基因表达水平。结果 100、200μmol/L Ost组可明显抑制Y1细胞增殖,且对含0.1%血清培养液中细胞抑制作用更明显。与阴性对照组比较,阳性对照组Y1细胞Star、Cyp11a1、Cyp21a1、Hsd3b2、Cyp11b1、Cyp17a1及Hsd17b3基因表达水平明显增强(P<0.05);干预24、48 h,50μmol/L Ost组Y1细胞皮质酮水平明显升高(P<0.05)。与24 h比较,干预48 h后,25、50μmol/L Ost组皮质酮含量明显升高(P<0.01)。干预24 h后,25、50μmol/L Ost组Star、Cyp21a1、Hsd3b2基因表达明显增强(P<0.05);干预48 h后,Star基因表达水平进一步增强(P<0.05),但Cyp11a1基因表达无明显差异(P>0.05)。干预24、48 h后,10、25、50μmol/L Ost组皮质酮合成酶Cyp11b1和性激素合成酶Cyp17a1基因表达明显增强(P<0.05);Ost对醛固酮合成酶Cyp11b2和性激素合成酶Hsd17b3基因表达水平未见明显作用。结论蛇床子素通过增强类固醇激素合成相关酶基因表达,参与调节肾上腺皮质功能,并以促进皮质酮的合成与分泌作用为主。Objective To study the effect of osthole（Ost） on adrenocortical function in Y1 mouse adreno-cortical tumor cells. Methods Y1 mouse adrenocortical tumor cells were taken as subjects in this experiment. In10. 0%, 1. 0%, and 0. 1% serum DMEM-F12 medium, Y1 cells were treated with 1, 10, 25, 50, 100, and 200μmol/L Ost for 24 and 48 h. 0. 1% Dimethyl Sulfoxide（DMSO） was taken as negative control group and 1 mmol/L（Bu） 2c AMP as positive control group. Cell growth morphology was observed under inverted microscope. Con-tents of corticosterone were tested by ELISA. Expression levels of steroids synthase such as Star, Cyp11a1,Cyp21a1, Hsd3b2, Cyp11b1, Cyp11b2, Cyp17a1, and Hsd17b3 m RNA were detected by Real time quantitative PCR（RT-q PCR）. Results Y1 cell proliferation was obviously inhibited by 100 and 200 μmol/L Ost, and its inhibitory effect was more significant in 0. 1% serum medium. Compared with the negative control group, gene expres-sions of Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1, and Hsd17b3 were significantly enhanced in the positive control group（P〈0. 05）. Y1 cell corticosterone levels significantly increased in 50 μmol/L Ost treatment group after 24-and 48-h intervention（P〈0. 05）. Contents of corticosterone increased more obviously in 25 and50 μmol/L Ost treatment groups after 48-h intervention, as compared with 24-h intervention（P〈0. 01）. After 24-h intervention, expression levels of Star, Cyp21a1, and Hsd3b2 genes were significantly up-regulated in 25 and50 μmol/L Ost groups（P〈0. 05）. Star gene expression was further enhanced after 48-h intervention（P〈0. 05）.However, Ost showed no effect on Cyp11a1（P〈0. 05）. Additionally, gene expressions of Cyp11b1 and Cyp17a1 were significantly enhanced by 10, 25, and 50 μmol/L Ost after treatment for 24 and 48 h（P〈0. 05）. Ost showed no obvious effect on Cyp11b2 and Hsd17b3 expressions. Conclusion Ost could regulate adrenal cortex function and promote corticosterone synthesis and secretion through s

关 键 词：蛇床子素 肾上腺皮质 Y1小鼠肾上腺皮质瘤细胞 11β-羟化酶1 20-碳链裂解酶 3β-羟基类固醇脱氢酶 类固醇急性调节蛋白 

分 类 号：R285.5[医药卫生—中药学]