Medical School, Institute of Reproductive Medicine, Nantong University, Nantong 226001, Jiangsu, China.
The Mary M. Wohlford Laboratory for Male Contraceptive Research, Center for Biomedical Research, Population Council, New York, USA.
Bioorg Chem. 2020 Nov;104:104295. doi: 10.1016/j.bioorg.2020.104295. Epub 2020 Sep 19.
Two synthesized resveratrol analogs from our laboratory, namely pinosylvin (3,5-dihydroxy-trans-stilbene, PIN) and 4,4'-dihydroxystilbene (DHS), have been carefully evaluated for treatment of oligoasthenospermia. Recent studies have demonstrated that PIN and DHS improved sperm quality in the mouse. However, the mechanism of action of PIN and DHS on oligoasthenospermia remains unknown. Herein, we investigated the mechanistic basis for improvements in sperm parameters by PIN and DHS in a mouse model of oligoasthenospermia induced by treatment with busulfan (BUS) at 6 mg/kg b.w.. Two weeks following busulfan treatment, mice were administered different concentrations of PIN or DHS daily for 2 consecutive weeks. Thereafter, epididymal sperm concentration and motility were determined, and histopathology of the testes was performed. Serum hormone levels including testosterone (T), luteinizing hormone (LH), and follicle stimulating hormone (FSH) were measured using corresponding specific enzyme-linked immunosorbent assay (ELISA) kits. Testicular mRNA expression profiles were determined by RNA sequencing analysis. These findings were validated by quantitative real-time PCR, western blotting and ELISA. Both PIN and DHS improved the epididymal sperm concentration and motility, enhanced testosterone levels, and promoted testicular morphological recovery following BUS treatment. PIN treatment was found to significantly reduce oxidative stress via the nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE)-dependent antioxidant, glutathione peroxidase 3. DHS treatment significantly reduced oxidative stress via the Nrf2-ARE-dependent antioxidants glutathione S-transferase theta 2 and glutathione S-transferase omega 2. In summary, PIN and DHS ameliorated oligoasthenospermia in this mouse model by attenuating oxidative stress via the Nrf2-ARE pathway.
我们实验室合成了两种白藜芦醇类似物，即松脂素（3,5-二羟基反式-芪，PIN）和 4,4'-二羟基二苯乙烯（DHS），并对其治疗少精子症进行了仔细评估。最近的研究表明，PIN 和 DHS 可改善小鼠的精子质量。然而，PIN 和 DHS 治疗少精子症的作用机制尚不清楚。在此，我们研究了 PIN 和 DHS 改善布硫磷（BUS，6mg/kg bw）诱导的少精子症小鼠精子参数的机制基础。BUS 处理 2 周后，每天给予不同浓度的 PIN 或 DHS 连续 2 周。然后，测定附睾精子浓度和活力，并进行睾丸组织病理学检查。使用相应的特异性酶联免疫吸附测定（ELISA）试剂盒测定血清激素水平，包括睾酮（T）、黄体生成素（LH）和卵泡刺激素（FSH）。通过 RNA 测序分析测定睾丸 mRNA 表达谱。通过定量实时 PCR、western blot 和 ELISA 验证了这些发现。PIN 和 DHS 均可提高附睾精子浓度和活力，提高睾酮水平，并促进 BUS 处理后睾丸形态恢复。发现 PIN 治疗可通过核因子红细胞 2 相关因子 2（Nrf2）-抗氧化反应元件（ARE）依赖性抗氧化剂谷胱甘肽过氧化物酶 3 显著降低氧化应激。DHS 治疗可通过 Nrf2-ARE 依赖性抗氧化剂谷胱甘肽 S-转移酶 theta 2 和谷胱甘肽 S-转移酶 omega 2 显著降低氧化应激。总之，PIN 和 DHS 通过 Nrf2-ARE 通路减轻氧化应激，改善了该小鼠模型的少精子症。
