附子肉桂抑制小鼠肾上腺皮质瘤细胞皮质酮生成的研究

Inhibitory effect of aconite and cinnamon on corticosterone production in Y1 mouse adrenocortical tumor cells

目的研究温热散寒中药药对"附子肉桂"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,设置空白对照组,1μmol/L佛司可林(FSK)组,以及不同浓度附子肉桂水煎醇沉液组(0.25 g/L、1 g/L、4 g/L、8 g/L),干预Y1细胞36 h或48 h。分别采用MTT检测细胞增殖,运用ELISA检测细胞分泌皮质酮含量,RT-qPCR检测类固醇激素合成酶基因表达,Western blot检测蛋白表达。结果与对照组(Con)比较,0.25 g/L、1 g/L、4 g/L、8 g/L附子肉桂组Y1细胞活力被显著抑制(P<0.05);4 g/L与8 g/L附子肉桂组Y1细胞皮质酮分泌显著下降(P<0.01);0.25 g/L、1 g/L、4 g/L、8 g/L附子肉桂组Cyp11a1、Cyp21a1、Cyp11b1基因表达显著下调(P<0.01),而8 g/L附子肉桂促进Star基因表达(P<0.01),且4 g/L附子肉桂显著抑制CYP11A1、CYP21A2、CYP11B1蛋白表达。与FSK组比较,4 g/L附子肉桂抑制Star、Cyp11a1、Cyp21a1、Cyp11b1基因表达后不能再被FSK诱导表达(P<0.01)。结论附子肉桂通过抑制细胞活力和下调类固醇激素合成酶表达,以抑制肾上腺皮质功能。

Objective To study the effect of aconite and cinnamon,two traditional Chinese medicines which have the function of dispelling cold by their warming and heating property,on the function of adrenal cortex.Methods Y1 mouse adrenocortical tumor cells(Y1 cells)were taken as subjects in this experiment.Y1 cells were treated with 0.25 g/L,1 g/L,4 g/L,8 g/L concentration of aconite and cinnamon decoction alcohol precipitation for 36 h or 48 h,and a blank control group and 1μmol/L FSK group were established.The cell proliferation was detected by MTT,the content of corticosterone secreted by the cells was detected by ELISA,the expression of steroid hormone synthase gene by real-time quantitative PCR(RT-qPCR)and protein expression by Western blot.Results Compared with the control group,the viability of Y1 cells was inhibited significantly in 0.25 g/L,1 g/L,4 g/L and 8 g/L aconite and cinnamon groups(P<0.05).The corticosterone secretion of Y1 cells was decreased significantly in 4 g/L and 8 g/L aconite and cinnamon groups(P<0.01).Cyp11 a1,Cyp21 a1 and Cyp11 b1 genes expression were inhibited significantly by 0.25 g/L,1 g/L,4 g/L and 8 g/L aconite and cinnamon(P<0.01).However,8 g/L aconite and cinnamon significantly promoted Star gene expression(P<0.01).And 4 g/L aconite and cinnamon significantly inhibited CYP11 A1,CYP21 A2,CYP11 B1 proteins expression.Compared with the FSK group,4 g/L aconite and cinnamon silenced the expression of Star,Cyp11 a1,Cyp21 a1,Cyp11 b1 and could not be induced by FSK(P<0.01).Conclusion Aconite and cinnamon inhibits adrenocortical function by inhibiting cell viability and down-regulating the expression of steroid hormone synthase.