1.C-Type Natriuretic Peptide/Natriuretic Peptide Receptor 2 Is Involved in Cell Proliferation and Testosterone Production in Mouse Leydig Cells
Lei YANG ; Lanjie LEI ; Qihan ZHAO ; Ying GONG ; Gaopeng GUAN ; Shaoxin HUANG
The World Journal of Men's Health 2019;37(2):186-198
PURPOSE: This study investigated the role of natriuretic peptide receptor 2 (NPR2) on cell proliferation and testosterone secretion 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 monophosphate production, testosterone secretion, cell proliferation, cell cycle and cell apoptosis in mouse Leydig cells were analyzed 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 secretion in mouse Leydig cells. CONCLUSIONS: Our study provides more evidences to better understand the function of CNP/NPR2 pathway in male reproduction, which may help us to treat male infertility.
Animals
;
Apoptosis
;
Blotting, Western
;
Cell Count
;
Cell Cycle
;
Cell Cycle Checkpoints
;
Cell Proliferation
;
Enzyme-Linked Immunosorbent Assay
;
Flow Cytometry
;
Germ Cells
;
Guanosine Monophosphate
;
Humans
;
Infertility, Male
;
Lentivirus
;
Leydig Cells
;
Male
;
Mice
;
Natriuretic Peptide, C-Type
;
Polymerase Chain Reaction
;
Receptors, Peptide
;
Reproduction
;
Reverse Transcription
;
RNA Interference
;
S Phase
;
Testicular Diseases
;
Testis
;
Testosterone