Assessment of Platelet-Rich Plasma (PRP) Effects on the Proliferation and Apoptosis of Cyclophosphamide-Induced Damage in Human Granulosa Cells

Abstract

Background: Chemotherapy-induced premature ovarian failure is a prevalent issue for cancer patients, and current treatment options are associated with limitations. Platelet-rich plasma (PRP) has emerged as a safe and straightforward treatment option for various medical conditions, including infertility. The study aims to assess the impact of PRP on the follicular function of granulosa cells damaged with Cyclophosphamide (CTX). Methods: The study evaluated the biological characteristics of Human Granulosa Cells (HGrC1) that were damaged with 4 hydroxy-cyclophosphamide (4-HC) as the active form of CTX. Then, cells were treated with different concentrations of platelet-rich plasma (PRP), and the expression of PI3K, K-RAS, BCL-2, P27, and Caspase 3 genes were analyzed. Cell viability and apoptosis were also assessed. Results: In the MTT assay, it was observed that cells treated with Cyclophosphamide had a faster growth rate when exposed to high concentrations of PRP. The results of the apoptosis assay using Annexin-V and propidium iodide (PI) revealed that PRP effectively inhibited apoptosis and enhanced cell proliferation across all incubation periods (24, 48, and 72 hours). Compared to untreated cells, those treated with different concentrations of PRP showed significantly lower levels of apoptosis, indicating higher cell viability. Real-time PCR results demonstrated that PRP treatment decreased the expression of apoptotic factors P27 and caspase 3 while increasing the expression of genes that promote cell survival and proliferation, such as BCL-2, PI3K, and K-RAS. These findings are consistent with previous research, which suggests that the growth factors present in platelets have anti-apoptotic effects and promote high rates of cell proliferation. Conclusion: The study suggests that PRP therapy may have potential benefits in promoting follicular growth and repairing ovarian tissue. However, further research is necessary to establish novel therapeutic strategies based on PRP therapy for treating infertility.

Keywords

• 4-hydroxycyclophosphamide
• cyclophosphamide
• platelet-rich plasma (prp)
• premature ovarian failure