Reproductive Medicine Center, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, People's Republic of China; Guangdong Provincial Key Laboratory of Reproductive Medicine, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, People's Republic of China.
Reproductive Medicine Center, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, People's Republic of China; Guangdong Provincial Key Laboratory of Reproductive Medicine, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, People's Republic of China.
F S Sci. 2024 Feb;5(1):16-23. doi: 10.1016/j.xfss.2023.10.005. Epub 2023 Oct 29.
To investigate the direct effect of growth differentiation factor 9 (GDF9) on androgen production in human theca cells.
Experimental study.
Tertiary hospital-based research laboratory.
PATIENT(S): Women who underwent in vitro fertilization and intracytoplasmic sperm injections at our clinic were included in this study.
INTERVENTION(S): Primary cultured human theca cells from women undergoing in vitro fertilization and intracytoplasmic sperm injection treatment were treated with GDF9, an activin receptor-like kinase 5 (ALK5) inhibitor, and a SMAD4 agonist.
MAIN OUTCOME MEASURE(S): The expression of androgen synthesis-related genes StAR, CYP17A1, and LHCGR, levels of androstenedione and testosterone, phosphorylation of SMAD2/3, and the interaction between bone morphogenic protein-activated type II receptor and ALK5 were evaluated using reverse transcription-quantitative polymerase chain reaction, Western blot, enzyme-linked immunosorbent assays, and coimmunoprecipitation assays, respectively.
RESULT(S): Growth differentiation factor 9 decreased StAR, CYP17A1, and LHCGR expression levels in human theca cells, which was prevented by treatment with the ALK5 inhibitor, and suppressed production of androgen in human theca cells. Growth differentiation factor 9 increased SMAD2/3 phosphorylation, and the ALK5 inhibitor also suppressed this effect. Bone morphogenic protein-activated type II receptor and ALK5 bound to each other after GDF9 stimulation. The SMAD4 agonist kartogenin also decreased messenger RNA levels of StAR and CYP17A1 and protein levels of StAR in human theca cells.
CONCLUSION(S): Growth differentiation factor 9 can activate the bone morphogenic protein-activated type II receptor-ALK5-SMAD2/3 signaling pathway, suppress CYP17A1 expression, and decrease androgen production in human theca cells.
研究生长分化因子 9（GDF9）对人卵巢膜细胞雄激素生成的直接作用。
实验研究。
我们诊所的基于三级医院的研究实验室。
纳入本研究的患者为在我们诊所接受体外受精和胞浆内精子注射的女性。
用 GDF9、激活素受体样激酶 5（ALK5）抑制剂和 SMAD4 激动剂处理接受体外受精和胞浆内精子注射治疗的人卵巢膜细胞原代培养物。
采用逆转录定量聚合酶链反应、Western blot、酶联免疫吸附试验和共免疫沉淀试验分别评估雄激素合成相关基因 StAR、CYP17A1 和 LHCGR 的表达、雄烯二酮和睾酮水平、SMAD2/3 的磷酸化以及骨形态发生蛋白激活型 II 受体与 ALK5 的相互作用。
GDF9 降低了人卵巢膜细胞中 StAR、CYP17A1 和 LHCGR 的表达水平，这种作用可被 ALK5 抑制剂阻断，并抑制人卵巢膜细胞中雄激素的产生。GDF9 增加了 SMAD2/3 的磷酸化，ALK5 抑制剂也抑制了这种作用。GDF9 刺激后，骨形态发生蛋白激活型 II 受体和 ALK5 相互结合。SMAD4 激动剂 kartogenin 也降低了 StAR 和 CYP17A1 的信使 RNA 水平和 StAR 的蛋白水平。
GDF9 可激活骨形态发生蛋白激活型 II 受体-ALK5-SMAD2/3 信号通路，抑制 CYP17A1 的表达，减少人卵巢膜细胞中的雄激素生成。
