Background: Despite its anticancer activity, cisplatin exhibits severe testicular toxicity when used in chemotherapy. Owing to its wide application in cancer therapy, the reduction of damage to normal tissue is of imminent clinical need. In this study, we evaluated the effects of catechin hydrate, a natural flavon-3-ol phytochemical, on cisplatin-induced testicular injury. Methods: Type 2 mouse spermatogonia (GC-1 spg cells) were treated with 0-100 μM catechin and cisplatin. Cell survival was estimated using a cell proliferation assay and Ki-67 immunostaining. Apoptosis was assessed via flow cytometry with the Dead Cell Apoptosis assay. To determine the antioxidant effects of catechin hydrate, Nrf2 expression was measured using qPCR and CellROX staining. The anti-inflammatory effects were evaluated by analyzing the gene and protein expression levels of iNOS and COX2 using qPCR and immunoblotting. Results: The 100 μM catechin hydrate treatment did not affect healthy GC-1 spg cells but, prevented cisplatin-induced GC-1 spg cell death via the regulation of anti-oxidants and inflammation-related molecules. In addition, the number of apoptotic cells, cleaved-caspase 3 level, and BAX gene expression levels were significantly reduced by catechin hydrate treatment in a cisplatin-induced GC-1 spg cell death model. In addition, antioxidant and anti-inflammatory marker genes, including Nrf2 , iNOS, and COX2 were significantly downregulated by catechin hydrate treatment in cisplatintreated GC-1 cells. Conclusions: Our study contributes to the opportunity to reintroduce cisplatin into systemic anticancer treatment, with reduced testicular toxicity and restored fertility.

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
    1. Cell culture and treatment
    2. Cell proliferation assay
    3. Immunostaining
    4. Flow cytometry analysis
    5. RNA extraction and quantitative PCR
    6. Immunoblotting
    7. Statistical analysis
RESULTS
    1. Catechin hydrate protects against cisplatin-inducedgerm cell damage
    2. Pro-apoptotic protein expression levels in GC-1spg cells after cispatin and catechin treatment
    3. Antioxidant and anti-inflammatory effects ofcatechin hydrate in cisplatin-induced GC-1 spg celldamage
DISCUSSION
CONCLUSION
REFERENCES

• Hyeon Woo Shim(Department of Animal Biotechnology, College of Life Science, Sangji University, Wonju 26339, Korea)
• Won-Yong Lee(Department of Livestock, Korea National University of Agriculture and Fisheries, Jeonju 54874, Korea)
• Youn-Kyung Ham(Department of Animal Science, College of Life Science, Sangji University, Wonju 26339, Korea)
• Sung Don Lim(Department of Plant Life and Resource Science, College of Life Science, Sangji University, Wonju 26339, Korea)
• Sun-Goo Hwang(Department of Plant Life and Resource Science, College of Life Science, Sangji University, Wonju 26339, Korea)
• Hyun-Jung Park(Department of Animal Biotechnology, College of Life Science, Sangji University, Wonju 26339, Korea) Corresponding author