C-Type Natriuretic Peptide/Natriuretic Peptide Receptor 2 Is Involved in Cell Proliferation and Testosterone Production in Mouse Leydig Cells

Abstract

Purpose: This study investigated the role of natriuretic peptide receptor 2 (NPR2) on cell proliferation and testosterone secre-tion in mouse Leydig cells.Materials and Methods: Mouse testis of different postnatal stages was isolated to detect the expression C-type natriuretic peptide (CNP) and its receptor NPR2 by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Leydig cells isolated from mouse testis were cultured and treated with shNPR2 lentiviruses or CNP. And then the cyclic guanosine mono-phosphate production, testosterone secretion, cell proliferation, cell cycle and cell apoptosis in mouse Leydig cells were ana-lyzed by ELISA, RT-qPCR, Cell Counting Kit-8, and flow cytometry. Moreover, the expression of NPR2, cell cycle, apoptosis proliferation and cell cycle related gene were detected by RT-qPCR and Western blot. Results: Knockdown of NPR2 by RNAi resulted in S phase cell cycle arrest, cell apoptosis, and decreased testosterone secre-tion in mouse Leydig cells.Conclusions: Our study provides more evidences to better understand the function of CNP/NPR2 pathway in male reproduc-tion, which may help us to treat male infertility.

Keywords

• Germ cells
• Leydig cells
• Testicular diseases
• Testosterone