烟碱对LPS诱导的兔子宫内膜上皮细胞炎症因子及前列腺素分泌的影响

doi:10.16431/j.cnki.1671-7236.2021.11.040

Abstract

Abstract: The purpose of this study was to investigate the mechanism of nicotine which is the high affinity agonist of α7nAChR on the inflammation of rabbit endometrial epithelial cells induced by lipopolysaccharides (LPS). Epithelial cells were isolated from the endometrium of rabbits in late estrus and stimulated with 100 ng/mL LPS for 12 h. CCK8 method was used to detect the effects of nicotine in 5, 10 and 20 μg/mL on cell survival rates, and appropriate concentrations of nicotine were screened for subsequent tests. The cells were divided into control group (CON), LPS group, LPS+nicotine group, LPS+nicotine+methyllycaconitine citrate(MLA, specific antagonist of α7nAChR) group. The contents of interleukin-1β(IL-1β), IL-6, IL-8, tumor necrosis factor-α(TNF-α), prostaglandin E₂(PGE₂) and prostaglandin F_2α(PGF_2α) in the supernatant of cell culture were detected by ELISA. Rabbit endometrial epithelial cells were successfully isolated and maintained good growth until the fifth generation. CCK8 test results showed that the survival rate of 20 μg/mL nicotine group was significantly decreased (P<0.05), and 10 μg/mL nicotine had no significant effect on the survival rate of cells(P>0.05), so 10 μg/mL nicotine was selected for the following test. ELISA results showed that compared with control group, the content of IL-1β, IL-6, IL-8, TNF-α, PGE₂ and PGF_2α was significantly increased in LPS group (P<0.05). When compared with LPS group, the content of IL-1β, IL-6, IL-8, TNF-α, PGE₂ and PGF_2α was significantly decreased in LPS+nicotine group(P<0.05), and the content of inflammatory factors and prostaglandins in LPS+nicotine+MLA group had no significant difference (P>0.05). These results suggested that nicotine could down-regulate the secretion of IL-1β, IL-6, IL-8, TNF-α, PGE₂ and PGF_2α in LPS-induced rabbit endometrial epithelial cells. It was speculated that nicotine played an anti-inflammatory role by down-regulating inflammatory factors and prostaglandins mediated by α7nAChR. These results might provide a reference for the selection of α7nAChR as a therapeutic target for endometritis.

Key words: α7nAChR; nicotine; inflammatory cytokines; prostaglandin

CLC Number:

S852.2

WEI Hongyu, BAO Zhaoxia, CHEN Hui, CAO Jinshan, HE Pengfei. Effects of Nicotine on LPS-induced Inflammatory Cytokines and Prostaglandin Secretion in Rabbit Endometrial Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4302-4310.

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Effects of Nicotine on LPS-induced Inflammatory Cytokines and Prostaglandin Secretion in Rabbit Endometrial Epithelial Cells

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