Department of Anesthesiology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, 325027, China.
Department of Pathology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, 325027, China.
Chem Biol Interact. 2019 Oct 1;312:108817. doi: 10.1016/j.cbi.2019.108817. Epub 2019 Sep 6.
Aconitine might have reproductive toxicity and the effects of aconitine on androgen synthesis in Leydig cells remain unclear. Here, we explore how aconitine affects androgen synthesis and metabolism in rat immature Leydig cells in vitro. Immature Leydig cells were isolated from 35-day-old male Sprague Dawley rats and cultured with 0-50 μM aconitine for 3 h in combination with LH, 8Br-cAMP, 22R-hydroxycholesterol, pregnenolone, progesterone, androstenedione, testosterone, and dihydrotestosterone, respectively. Medium androgens were measured. The levels of Leydig cell mRNAs, Lhcgr, Scarb1, Star, Cyp11a1, Hsd3b1, Cyp17a1, Hsd17b3, Srd5a1, and Akr1c14, were measured by qPCR. ROS and apoptosis were determined after 24-h aconitine treatment. Aconitine inhibited basal androgen production in Leydig cells at 0.05 μM and the higher concentrations. Aconitine blocked pregnenolone, progesterone, and androstenedione mediated androgen outputs without affecting 22R-hydroxycholesterol-mediated androgen production at 5 μM. Aconitine also inhibited LH and 8Br-cAMP stimulated androgen outputs at 5 μM. Further investigation showed that aconitine blocked androgen synthesis via down-regulating the expression of Scarb1, Hsd3b1, Cyp17a1, and Hsd17b3. At 50 μM, aconitine also induced ROS generation and increased apoptotic rate of Leydig cells. Aconitine lowered serum testosterone levels at 1.5 mg/kg after 7 days of oral exposure from postnatal day 35. In conclusion, aconitine inhibits androgen synthesis.
乌头碱可能具有生殖毒性，其对间质细胞雄激素合成的影响尚不清楚。在这里，我们研究了乌头碱如何影响体外培养的大鼠未成熟间质细胞的雄激素合成和代谢。从 35 日龄雄性 Sprague Dawley 大鼠中分离未成熟间质细胞，并与 LH、8Br-cAMP、22R-羟胆固醇、孕烯醇酮、孕酮、雄烯二酮、睾酮和二氢睾酮分别孵育 3h，用 0-50µM 乌头碱处理。测量培养基中的雄激素。用 qPCR 测量间质细胞 mRNA 的水平，Lhcgr、Scarb1、Star、Cyp11a1、Hsd3b1、Cyp17a1、Hsd17b3、Srd5a1 和 Akr1c14。用 24h 乌头碱处理后测定 ROS 和细胞凋亡。乌头碱在 0.05µM 时抑制基础雄激素的产生，浓度较高时抑制作用更强。乌头碱在 5µM 时阻断了 pregnenolone、孕酮和雄烯二酮介导的雄激素产物的形成，而不影响 22R-羟胆固醇介导的雄激素产物的形成。乌头碱还抑制了 5µM 时 LH 和 8Br-cAMP 刺激的雄激素产物的形成。进一步研究表明，乌头碱通过下调 Scarb1、Hsd3b1、Cyp17a1 和 Hsd17b3 的表达来阻断雄激素的合成。在 50µM 时，乌头碱还诱导了 ROS 的产生并增加了间质细胞的凋亡率。在口服暴露后第 35 天的 7 天内，1.5mg/kg 的乌头碱使血清睾酮水平降低。综上所述，乌头碱抑制雄激素的合成。
