UFL1 Alleviates LPS-Induced Apoptosis by Regulating the NF-κB Signaling Pathway in Bovine Ovarian Granulosa Cells.

Xinling Wang,Lian Li,Genlin Wang,Yiru Wang,Zhaoyu Han,Chengmin Li

doi:10.3390/biom10020260

Abstract

Ubiquitin-like modifier 1 ligating enzyme 1 (UFL1) is an E3 ligase of ubiquitin fold modifier 1 (UFM1), which can act together with its target protein to inhibit the apoptosis of cells. Lipopolysaccharides (LPS) can affect the ovarian health of female animals by affecting the apoptosis of ovarian granulosa cells. The physiological function of UFL1 on the apoptosis of bovine (ovarian) granulosa cells (bGCs) remains unclear; therefore, we focused on the modulating effect of UFL1 on the regulation of LPS-induced apoptosis in ovarian granulosa cells. Our study found that UFL1 was expressed in both the nucleus and cytoplasm of bGCs. The results here demonstrated that LPS caused a significant increase in the apoptosis level of bGCs in cows, and also dramatically increased the expression of UFL1. Furthermore, we found that UFL1 depletion caused a significant increase in apoptosis (increased the expression of BAX/BCL-2 and the activity of caspase-3). Conversely, the overexpression of UFL1 relieved the LPS-induced apoptosis. In order to assess whether the inhibition of bGCs apoptosis involved in the nuclear factor-κB (NF-κB) signaling pathway resulted from UFL1, we detected the expression of NF-κB p-p65. LPS treatment resulted in a significant upregulation in the protein concentration of NF-κB p-p65, and knockdown of UFL1 further increased the phosphorylation of NF-κB p65, while UFL1 overexpression significantly inhibited the expression of NF-κB p-p65. Collectively, UFL1 could suppress LPS-induced apoptosis in cow ovarian granulosa cells, likely via the NF-κB pathway. These results identify a novel role of UFL1 in the modulation of bGC apoptosis, which may be a potential signaling target to improve the reproductive health of dairy cows.

Highlights

The reproductive performance of dairy cows has attracted growing attention in the cattle industry [1,2]
Ubiquitin-like modifier 1 ligating enzyme 1 (UFL1) could suppress LPS-induced apoptosis in cow ovarian granulosa cells, likely via the nuclear factor-κB (NF-κB) pathway. These results identify a novel role of UFL1 in the modulation of bovine (ovarian) granulosa cells (bGCs) apoptosis, which may be a potential signaling target to improve the reproductive health of dairy cows
Follicle-stimulating hormone receptor (FSHR) immunocytochemical staining is the specific staining of ovarian granulosa cells, where the nucleus is dark blue, and the FSHR positive staining is localized in the cell membrane, which is stained green

Summary

Introduction

The reproductive performance of dairy cows has attracted growing attention in the cattle industry [1,2]. The ovaries are a key reproductive organ and play a crucial role in the animal production performance. Follicles are a basic constituent in the ovary, which is mainly composed of a single type of oocyte, named granulosa cells (GCs). Previous studies have indicated that follicular granulosa cells modulate follicular development through excreting estrogen and progesterone [5]. As key functional components of follicles, perform an initiating role in follicular atresia and further affect the reproductive facility [6,7,8]. Granulosa cell apoptosis appears to be an essential part of ovarian development, and it is a reflection of the mitogenic growth of the follicle [9]. Previous studies have elaborated that lipopolysaccharides (LPS) could induce an ovarian inflammatory response and affect ovarian development directly [10,11]

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Results

Conclusion