Oxycodone Alleviates Endometrial Injury via the TLR4/NF-κB Pathway

Abstract

Endometrial injury is a common female disease. This study was designed to illustrate the effects of oxycodone on mifepristone-induced human endometrial stromal cells (hEndoSCs) injury and delineate the underlying molecular mechanism. hEndoSCs were stimulated with mifepristone to generate the endometrial injury in vitro model. hEndoSCs viability, cytotoxicity, and apoptosis were measured by methyl thiazolyl tetrazolium (MTT) assay, lactate dehydrogenase assay (LDH), and flow cytometry (FCM) analysis, respectively. Meanwhile, quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot assay were conducted to evaluate gene and protein expressions. The secretions of inflammatory cytokines (TNF-α, IL-1β, and IL-6) were measured using enzyme-linked immunosorbent assay (ELISA). The data revealed that mifepristone exposure memorably inhibited hEndoSCs viability and promoted cell apoptosis and inflammatory cytokines secretion, and oxycodone had no cytotoxicity on hEndoSCs. Oxycodone increased hEndoSCs growth, blocked cell apoptosis, enhanced Bcl-2 expression, reduced Bax levels, and decreased the secretion of inflammatory cytokines in mifepristone-induced hEndoSCs, exhibiting the protective effects in endometrial injury. In addition, the TLR4/NF-κB pathway-related protein levels (TLR4 and p-p65) in mifepristone-treated hEndoSCs were enhanced, while these enhancements were inhibited by oxycodone treatment. In conclusion, oxycodone exhibited the protective role in mifepristone-triggered endometrial injury via inhibiting the TLR4/NF-κB signal pathway.

Oxycodone alleviates mifepristone‐stimulated human endometrial stromal cell injury by activating the Keap1/Nrf2/HO‐1 signaling pathway

“…1 Also, administration of mifepristone could induce the section of inflammatory cytokines (such as TNF-α, IL-β, or IL-6). 11,33 Thus, the levels of TNF-α, IL-β, and IL-6 in hEndoSCs were detected, and consistent with the previous study, 11 the findings indicated that mifepristone promoted TNF-α, IL-β, and IL-6 secretion in hEndoSCs. Oxycodone reduced inflammatory cytokine release in hEndoSCs in a dose-dependent manner, indicating that oxycodone abolished inflammatory cytokine release during mifepristone-triggered hEndoSC damage.…”

“…Endometrial injury is a common gynecological disease caused by a variety of factors, including endocrine disorders, inflammation, and abortion, and in severe cases, may cause infertility 1,28 . Our previous research found that oxycodone alleviates endometrial injury via the inhibition of the activation of the TLR4/NF‐kB pathway 11 . However, the function of oxycodone and its latent pathogenesis in endometrial injury have not been completely investigated.…”

“…Next, mifepristone (60 μmol/L, 48 h) was used to stimulate hEndoSCs to establish an endometrial injury model. 24 For oxycodone treatment, hEndoSCs were exposed to 1, 5, 10, 15, or 20 μg/mL oxycodone for 24 h. 11 To explore whether the Keap1/Nrf2/HO-1 signaling pathway was involved in the action of oxycodone on mifepristone-induced hEndoSCs, a Keap1/Nrf2/HO-1 pathway inhibitor (ML385) was used. Also, the mifepristone-induced hEndoSCs were treated with oxycodone (10 μg/mL) or/and 5 μM ML385 for 24 h.…”

“…Green et al 10 revealed that maternal oxycodone treatment led to changes in mouse placental histology. A recent research reported that oxycodone alleviates endometrial injury by inhibiting the activation of the TLR4/NF‐kB pathway 11 . However, the underlying molecular mechanisms under the effect of oxycodone against endometrial injury remain to be explored.…”

“…Mifepristone is a steroidal antiprogesterone drug, and it is widely used around the globe in the field of reproductive medicine 21 . Mifepristone‐induced damage of hEndoSCs has been confirmed on the basis of molecular, morphological, and functional features 11,22 . In recent years, mifepristone‐induced hEndoSCs have been used as an in vitro‐induced endometrial injury model 11,23,24 .…”