Kim Jin Hee, Lee Jae Hoon, Koo Yoon Jung, Song Jong Wook
Department of Anesthesiology and Pain Medicine, Yonsei University College of Medicine, 50 Yonsei- ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
Anesthesia and Pain Research Institute, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul, 03722, Republic of Korea.
BMC Mol Cell Biol. 2025 Apr 16;26(1):12. doi: 10.1186/s12860-025-00537-9.
Phthalates are plasticizers that cause inflammation in several cell types and adversely affect the health of humans and animals. Nacetylcysteine (NAC) has been shown to exert antioxidant effects in various diseases. However, the effect of NAC on diethyl phthalate (DEP)-induced toxicity in macrophages has not yet been elucidated. In this study, we investigated the effect and underlying mechanisms of NAC on DEP-induced inflammation in RAW264.7 macrophages. RAW264.7 macrophages were pretreated with NAC for 2 h followed by exposure to DEP. We investigated the effect of NAC on NO, reactive oxygen species (ROS), prostaglandin E2 (PGE2), and glutathione (GSH) levels following DEP exposure. In addition, pathway-related genes including cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), mitogen-activated protein kinase (MAPK), and signal transducer and activator of transcription (STAT) were evaluated using western blot.
Treatment with 100 and 300 µM DEHP, DBP, and DEP significantly increased the protein levels of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) compared with those in the control group. However, NAC pretreatment downregulated the levels of NO, PGE2, and ROS, elevated GSH levels, and suppressed the mRNA levels of inflammatory cytokines such as interleukin (IL)-1β, IL-6, COX-2, and iNOS compared with those in the DEP-treated group. In addition, NAC significantly reduced the levels of p-JNK and p-STAT1/3 in RAW264.7 macrophages treated with DEP.
NAC pretreatment inhibits DEP-induced inflammation via the MAPK/JNK and STAT1/3 pathways in macrophages.
邻苯二甲酸盐是增塑剂，可在多种细胞类型中引发炎症，并对人类和动物的健康产生不利影响。已证明N-乙酰半胱氨酸（NAC）在各种疾病中发挥抗氧化作用。然而，NAC对巨噬细胞中邻苯二甲酸二乙酯（DEP）诱导的毒性作用尚未阐明。在本研究中，我们调查了NAC对RAW264.7巨噬细胞中DEP诱导的炎症的影响及其潜在机制。RAW264.7巨噬细胞先用NAC预处理2小时，然后暴露于DEP。我们研究了NAC对DEP暴露后一氧化氮（NO）、活性氧（ROS）、前列腺素E2（PGE2）和谷胱甘肽（GSH）水平的影响。此外，使用蛋白质印迹法评估了包括环氧化酶-2（COX-2）、诱导型一氧化氮合酶（iNOS）、丝裂原活化蛋白激酶（MAPK）和信号转导和转录激活因子（STAT）在内的通路相关基因。
与对照组相比，用100和300µM的邻苯二甲酸二（2-乙基己基）酯（DEHP）、邻苯二甲酸二丁酯（DBP）和DEP处理显著增加了环氧化酶-2（COX-2）和诱导型一氧化氮合酶（iNOS）的蛋白质水平。然而，与DEP处理组相比，NAC预处理下调了NO、PGE2和ROS的水平，提高了GSH水平，并抑制了白细胞介素（IL）-1β、IL-6、COX-2和iNOS等炎性细胞因子的mRNA水平。此外，NAC显著降低了DEP处理的RAW264.7巨噬细胞中p-JNK和p-STAT1/3的水平。
NAC预处理通过巨噬细胞中的MAPK/JNK和STAT1/3通路抑制DEP诱导的炎症。
