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Apigenin Isolated from Carduus crispus Protects against H2O2-Induced Oxidative Damage and Spermatogenic Expression Changes in GC-2spd Sperm Cells
Authors:Spandana Rajendra Kopalli  Sung-Kwang Yoo  Bokyung Kim  Si-Kwan Kim  Sushruta Koppula
Institution:1.Department of Bioscience and Biotechnology, Sejong University, Gwangjin-gu, Seoul 05006, Korea;2.Ottugi Food Co., Ltd., Anyang-si 14060, Gyeonggi-do, Korea;3.Department of Physiology and Immunology, School of Medicine, Konkuk University, Chungju 27381, Korea;4.Department of Integrated Biosciences, College of Biomedical & Health Science, Konkuk University, Chungju 27381, Korea;
Abstract: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.
Keywords:welted thistle  apigenin  hydrogen peroxide  reactive oxygen species  testicular cells  nectin  CREB
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