Department of Prosthodontics, Hospital of Stomatology Affiliated to Southwest Medical University, Luzhou, Sichuan 646000, P.R. China.
Mol Med Rep. 2020 Oct;22(4):3299-3306. doi: 10.3892/mmr.2020.11433. Epub 2020 Aug 13.
N-acetyl cysteine (NAC) has been used to inhibit lipopolysaccharide (LPS)-induced inflammation. However, the molecular mechanism underlying its anti‑inflammatory effects remains to be elucidated. The present study aimed to determine the effect of NAC on the LPS‑induced inflammatory response in bone marrow mesenchymal stem cells (BMSCs) and elucidate the underlying molecular mechanism. First, BMSCs were stimulated by LPS following pretreatment with NAC (0, 0.1, 0.5, 1 or 2 mM). A Cell Counting Kit 8 assay was used to determine the number of viable cells and 1 mM NAC was selected as the experimental concentration. Then, the secretion of inflammatory factors, including interleukin (IL)‑1β, IL‑6 and tumor necrosis factor‑α was evaluated by enzyme‑linked immunosorbent assay. Finally, the expression levels of mRNA and proteins, including apoptosis‑associated speck‑like protein containing a CARD (ASC), nucleotide‑binding oligomerization domain‑like receptor protein 3 (NLRP3), caspase‑1, thioredoxin‑interacting protein (TXNIP), and thioredoxin (TRX), were evaluated by reverse transcription‑quantitative PCR and western blot analysis, respectively. The results demonstrated that the secretion of inflammatory factors, which was increased by the administration of LPS, was reduced by pretreatment with NAC. Furthermore, NAC reduced the expression of ASC, NLRP3, caspase‑1 and TXNIP, but enhanced that of TRX. To conclude, NAC had anti‑inflammatory effects on LPS‑stimulated BMSCs, which was closely associated with the TXNIP/NLRP3/IL‑1β signaling pathway. Thus, NAC may be a promising treatment to attenuate the inflammatory response in LPS‑induced BMSCs.
N-乙酰半胱氨酸 (NAC) 已被用于抑制脂多糖 (LPS) 诱导的炎症。然而，其抗炎作用的分子机制仍有待阐明。本研究旨在确定 NAC 对 LPS 诱导的骨髓间充质干细胞 (BMSC) 炎症反应的影响，并阐明其潜在的分子机制。首先，用 LPS 刺激 BMSC，然后用 NAC（0、0.1、0.5、1 或 2 mM）预处理。使用细胞计数试剂盒 8 测定活细胞数，并选择 1 mM NAC 作为实验浓度。然后，通过酶联免疫吸附试验评估炎症因子（包括白细胞介素 (IL)-1β、IL-6 和肿瘤坏死因子-α）的分泌。最后，通过逆转录-定量 PCR 和 Western blot 分析分别评估 mRNA 和蛋白的表达水平，包括凋亡相关斑点样蛋白含 CARD（ASC）、核苷酸结合寡聚化结构域样受体蛋白 3（NLRP3）、半胱天冬酶-1、硫氧还蛋白相互作用蛋白 (TXNIP) 和硫氧还蛋白 (TRX)。结果表明，LPS 给药增加的炎症因子分泌被 NAC 预处理所减少。此外，NAC 降低了 ASC、NLRP3、半胱天冬酶-1 和 TXNIP 的表达，但增强了 TRX 的表达。总之，NAC 对 LPS 刺激的 BMSC 具有抗炎作用，这与 TXNIP/NLRP3/IL-1β 信号通路密切相关。因此，NAC 可能是一种有前途的治疗方法，可减轻 LPS 诱导的 BMSC 中的炎症反应。
