Abstract
BackgroundLeydig cells reflect the activation of inflammation, decrease of androgen production, inhibition of cell growth and promotion of cell apoptosis under orchitis. Maternally expressed gene 3 (MEG3) exerts a crucial role in various human diseases, but under orchitis, the role and underlying molecular mechanism of MEG3 in Leydig cells remain unclear.MethodsLipofectamine 2000 was used for the cell transfections. qPCR and western blots assay were applied to assess the gene expression. ELISA assay was used to measure the TNFα, IL6 and testosterone secretion. CCK8 and EdU assay was employ to test the cell viability and proliferation respectively. Luciferase reporter and RIP assay were introduced to detect the binding of miR-93-5p with MEG3 and PTEN.ResultsLipopolysaccharides (LPS) induced TNFα and IL6 secretion, lowered testosterone production, inhibited cell viability and proliferation, and induced cell apoptosis in Leydig cells. MEG3 was upregulated in Leydig cells treated with LPS and that knockdown of MEG3 inhibited the role of LPS in Leydig cells. MEG3 absorbed miR-93-5p and that suppression of miR-93-5p restored the role of silenced MEG3 in Leydig cells under LPS treatment. miR-93-5p inhibited PTEN expression and that over-expressed PTEN alleviated the effect of miR-93-5p in Leydig cells treated with LPS. LPS activated the MEG3/miR-93-5p/PTEN signalling pathway in Leydig cells.ConclusionsThis study revealed that MEG3 serves as a molecular sponge to absorb miR-93-5p, thus leading to elevation of PTEN expression in Leydig cells under LPS treatment, offering a theoretical basis on which to establish potential new treatment strategies for orchitis.
Highlights
As a dominant cause of infertility, orchitis is defined as inflammation of one or both testicles [1]
Silencing of Maternally expressed gene 3 (MEG3) attenuated the role of LPS in Leydig cells To investigate whether MEG3 was involved in the role of LPS in Leydig cells, we measured the expression of MEG3 in Leydig cells treated with different concentrations of LPS
The results of qPCR analysis demonstrated that both shMEG3s could efficiently downregulate the expression of MEG3 in Leydig cells, and shMEG3–1 was chosen for further experiments (Fig. 2b)
Summary
As a dominant cause of infertility, orchitis is defined as inflammation of one or both testicles [1]. Zhou et al Reproductive Biology and Endocrinology (2021) 19:33 and testosterone production in Leydig cells might aid in development of innovative strategies for the treatment of orchitis. Once considered “transcriptional noise”, lncRNAs play an important role in a variety of cellular progresses, including chromatin remodelling, cell proliferation and cycling, cell death, metastasis, development and tumourigenesis via regulation of genes at the transcriptional, post-transcriptional and translational levels [10,11,12,13]. Leydig cells reflect the activation of inflammation, decrease of androgen production, inhibition of cell growth and promotion of cell apoptosis under orchitis. Expressed gene 3 (MEG3) exerts a crucial role in various human diseases, but under orchitis, the role and underlying molecular mechanism of MEG3 in Leydig cells remain unclear
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