内质网应激预适应对LPS诱导的山羊子宫内膜上皮细胞炎性反应的保护作用

doi:10.11843/j.issn.0366-6964.2023.08.038

Abstract

Abstract: We attempted to investigate the role of endoplasmic reticulum stress (ERS) in lipopolysaccharide (LPS)-induced inflammatory responses in goat endometrial epithelial cells (gEECs) and provide a preliminary basis for elucidating the mechanism of ERS in ruminant endometritis. In this study, gEECs were treated with ERS activator tunicamycin (TM) and inhibitor 4-phenylbutyric acid (4-PBA) separately to activate and inhibit ERS, and RT-qPCR and Western blot techniques were used to detect the effect of ERS on inflammation-related genes expression. Next, gEECs were pretreated with TM and 4-PBA, respectively, and then treated with E.coli LPS to induce inflammatory responses in gEECs, the effects and mechanisms of ERS on the inflammatory responses induced by LPS in gEECs were detected by RT-qPCR and Western blot. The results showed that activation of ERS by TM treatment significantly inhibited the expression of inflammatory cytokines IL-6 and TNF-α mRNA in gEECs (P<0.05). The mRNA expression levels of classical inflammatory pathway-related genes TLR4, NF-κB P65, and NLRP3 was also significantly suppressed (P<0.01). Meanwhile, it also significantly inhibited the phosphorylation of NF-κB P65 protein and the expression of NLRP3 protein (P<0.05). However, 4-PBA treatment significantly promoted the expression of inflammation-related genes (IL-6, TNF-α, TLR4, NF-κB P65, and NLRP3) in gEECs (P<0.05). Further study revealed that TM pretreatment of gEECs significantly inhibited the expression of LPS-induced inflammatory cytokine IL-6 mRNA (P<0.05). It also significantly inhibited LPS-induced TLR4, NF-κB P65, and NLRP3 mRNA expression as well as phosphorylated NF-κB P65 protein and NLRP3 protein (P<0.05). 4-PBA pretreatment of gEECs then significantly promoted the LPS-induced inflammatory response in gEECs (P<0.05). The above results suggest that ERS alleviates LPS-induced inflammatory response in gEECs by inhibiting the activation of NF-κB pathway and NLRP3 inflammatory vesicle pathway. These results provide a pre-theoretical basis for further elucidation of the development of goat endometritis.

Key words: endoplasmic reticulum stress, endometritis, lipopolysaccharide, NF-κB pathway, NLRP3 inflammatory vesicles

CLC Number:

S857.2

GAO Kangkang, YI Yanyan, ZHAO Yiteng, LIN Pengfei, CHEN Huatao, JIN Yaping. Protective Effect of Endoplasmic Reticulum Stress Preadaptation on LPS-Induced Inflammatory Response in Goat Endometrial Epithelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3546-3556.

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Protective Effect of Endoplasmic Reticulum Stress Preadaptation on LPS-Induced Inflammatory Response in Goat Endometrial Epithelial Cells

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