Hu Xiaoxuan, Zhang Na, Zhong Yuxu, Liu Tao, Zhu Xiaoji
Weifang No. 2 People's Hospital, Weifang Respiratory Disease Hospital, Weifang, China.
State Key Laboratory of Antitoxic Drugs and Toxicology, Institute of Toxicology and Pharmacology, Academy of Military Medical Sciences, Beijing, China.
Int J Toxicol. 2025 Jul-Aug;44(4):314-327. doi: 10.1177/10915818251315907. Epub 2025 Jan 31.
Sulfur mustard (SM) is a highly toxic bifunctional alkylating agent that inflicts severe damage on the respiratory tract. Although numerous studies have examined the mechanisms underlying SM-induced pulmonary injury, the exact pathways involved remain unclear. This study aims to investigate an acute pulmonary injury model, with SM administered as a single intraperitoneal injection (8 mg/kg) or single intratracheal instillation (2 mg/kg) at equal toxicity doses (1LD50). The results revealed that epithelial cells in the alveolar septa of the intraperitoneal SM group exhibited a significantly higher expression of apoptotic markers, including pro-apoptotic protein Bax, caspase-3, and caspase-9 proteins, than those in the tracheal SM group. Conversely, the expression of the anti-apoptotic protein Bcl-2 was significantly lower in the intraperitoneal SM group than in the tracheal SM group, as confirmed by TUNEL staining and immunohistochemical staining. The intraperitoneal SM group exhibited markedly higher expression of fibrosis-related proteins, including MMP-2, MMP-9, TIMP-1, TIMP-2, collagen type I, collagen type III, TGF-β1, and Smad7, than the tracheal SM group. These markers, detected through immunohistochemical immunolabeling, indicate a more significant fibrotic response in the intraperitoneal group. In summary, this study demonstrates that intraperitoneal exposure to SM results in increased apoptosis, elevated expression of pro-apoptotic proteins, and fibrosis-related proteins in the alveolar epithelial cells compared with intratracheal exposure, even at equivalent toxicity levels. Our findings highlight the suitability of the intraperitoneal route for further investigation and identify apoptotic and fibrosis-related proteins as potential targets for intervention in SM-induced pulmonary injury.
硫芥（SM）是一种剧毒的双功能烷基化剂，会对呼吸道造成严重损害。尽管众多研究已经探究了SM诱导肺损伤的潜在机制，但具体涉及的途径仍不清楚。本研究旨在探究一种急性肺损伤模型，以同等毒性剂量（1LD50）通过单次腹腔注射（8mg/kg）或单次气管内滴注（2mg/kg）给予SM。结果显示，腹腔注射SM组肺泡隔中的上皮细胞凋亡标志物表达显著高于气管内注射SM组，这些标志物包括促凋亡蛋白Bax、半胱天冬酶-3和半胱天冬酶-9蛋白。相反，经TUNEL染色和免疫组化染色证实，腹腔注射SM组抗凋亡蛋白Bcl-2的表达显著低于气管内注射SM组。腹腔注射SM组纤维化相关蛋白的表达明显高于气管内注射SM组，这些蛋白包括基质金属蛋白酶-2、基质金属蛋白酶-9、金属蛋白酶组织抑制因子-1、金属蛋白酶组织抑制因子-2、I型胶原、III型胶原、转化生长因子-β1和Smad7。通过免疫组化免疫标记检测到的这些标志物表明腹腔注射组的纤维化反应更显著。总之，本研究表明，即使在同等毒性水平下，与气管内暴露相比，腹腔暴露于SM会导致肺泡上皮细胞凋亡增加、促凋亡蛋白和纤维化相关蛋白表达升高。我们的研究结果突出了腹腔途径在进一步研究中的适用性，并确定凋亡和纤维化相关蛋白作为干预SM诱导肺损伤的潜在靶点。
