论文部分内容阅读
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 investigate the effect of Osthole (Ost) on the function of adrenal cortex. Methods Y1 mouse adrenocortical tumor cells were used as experimental subjects. Cells were cultured with 10%, 1% and 0.1% serum. The cells were treated with 1, 10, 25, 50, 100 and 200 μmol / 48 h, 0.1% dimethylsulfoxide (DMSO) as the negative control group, adenylyl cyclase activator (Bu) 2cAMP as the positive control group, the morphological changes of the cells were observed under an inverted microscope, The levels of corticosterone, corticosterone, Cyp11a1, Cyp21a1, Hsd3b2 and 11β-hydroxysteroid were assayed by RT-qPCR. (Cyp11b1), 11β-hydroxylase 2 (Cyp11b2), 17α-hydroxylase / 17,20-carbon chain cleaving enzyme (Cyp17a1) and 17β-hydroxysteroid dehydrogenase 3 (Hsd17b3). Results 100, 200μmol / L Ost group could significantly inhibit the proliferation of Y1 cells, and the inhibitory effect on the cells containing 0.1% serum was more obvious. Compared with the negative control group, the expression levels of Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1 and Hsd17b3 in Y1 cells in the positive control group were significantly increased (P <0.05) The level was significantly higher (P <0.05). Compared with 24 h, the levels of corticosterone in 25 and 50 μmol / L Ost groups were significantly increased after 48 h intervention (P <0.01). After intervention for 24 h, the expression of Star, Cyp21a1 and Hsd3b2 genes in 25 and 50μmol / L Ost groups were significantly increased (P <0.05), and the expression of Star gene was further enhanced 48h after intervention (P <0.05), but the expression of Cyp11a1 gene was not significant Difference (P> 0.05). After intervention for 24 and 48 h, the expression of corticosterone synthase Cyp11b1 and sex hormone synthase Cyp17a1 in 10, 25 and 50μmol / L Ost groups were significantly increased (P <0.05); Ost also inhibited the expression of aldosterone synthase gene Cyp11b2 and sex hormone synthase Hsd17b3 No significant effect. Conclusion Osthole can regulate adrenocortical function by enhancing the gene expression of enzymes involved in steroid hormone synthesis and promote the synthesis and secretion of corticosterone.