Department of Bioscience and Biotechnology, Sejong University, Gwangjin-gu, Seoul 05006, Korea.
Ottugi Food Co., Ltd., Anyang-si 14060, Gyeonggi-do, Korea.
Molecules. 2022 Mar 8;27(6):1777. doi: 10.3390/molecules27061777.
Testicular oxidative stress is one of the most common factors underlying male infertility. Welted thistle, Carduus crispus Linn., and its bioactive principles are attracting scientific interest in treating male reproductive dysfunctions. Here, the protective effects of apigenin isolated from C. crispus against oxidative damage induced by hydrogen peroxide (H2O2) and dysregulation in spermatogenesis associated parameters in testicular sperm cells was investigated. Cell viabilities, ROS scavenging effects, and spermatogenic associated molecular expressions were measured by MTT, DCF-DA, Western blotting and real-time RT-PCR, respectively. A single peak with 100% purity of apigenin was obtained in HPLC conditions. Apigenin treated alone (2.5, 5, 10 and 20 µM) did not exhibit cytotoxicity, but inhibited the H2O2-induced cellular damage and elevated ROS levels significantly (p < 0.05 at 5, 10 and 20 µM) and dose-dependently. Further, H2O2-induced down-regulation of antioxidant (glutathione S-transferases m5, glutathione peroxidase 4, and peroxiredoxin 3) and spermatogenesis-associated (nectin-2 and phosphorylated-cAMP response element-binding protein) molecular expression in GC-2spd cells were attenuated by apigenin at both protein and mRNA levels (p < 0.05). In conclusion, our study showed that apigenin isolated from C. crispus might be an effective agent that can protect ROS-induced testicular dysfunctions.
睾丸氧化应激是男性不育的最常见因素之一。皱叶酸模（Carduus crispus Linn.）及其生物活性成分在治疗男性生殖功能障碍方面引起了科学界的兴趣。在这里，研究了从皱叶酸模中分离出的芹菜素对过氧化氢（H2O2）诱导的氧化损伤和睾丸精子细胞中与精子发生相关的参数失调的保护作用。通过 MTT、DCF-DA、Western blot 和实时 RT-PCR 分别测量细胞活力、ROS 清除作用和与精子发生相关的分子表达。在 HPLC 条件下获得了具有 100%纯度的芹菜素单一峰。芹菜素单独处理（2.5、5、10 和 20 µM）不会表现出细胞毒性，但显著抑制 H2O2 诱导的细胞损伤并显著升高 ROS 水平（在 5、10 和 20 µM 时 p < 0.05）和剂量依赖性。此外，H2O2 诱导的 GC-2spd 细胞中抗氧化剂（谷胱甘肽 S-转移酶 m5、谷胱甘肽过氧化物酶 4 和过氧化物酶 3）和与精子发生相关的分子表达（ nectin-2 和磷酸化-cAMP 反应元件结合蛋白）下调被芹菜素在蛋白和 mRNA 水平上减弱（p < 0.05）。总之，我们的研究表明，皱叶酸模中分离出的芹菜素可能是一种有效的能够保护 ROS 诱导的睾丸功能障碍的药物。
