Lang Wuying, Zhang Jiayi, Xiao Xuejun, Cheng Min, Zheng Xin, Gong Haizhou, Ali Ihsan, Zhao Yongping, Jia Feng, Wang Zhe, Wang Jing, Li Wei, Zhang Haihua
College of Biology Pharmacy and Food Engineering, Shangluo University, Shangluo, China.
Department of Medical Oncology, Guangshan County People's Hospital, Xinyang, China.
Comb Chem High Throughput Screen. 2025 Jan 29. doi: 10.2174/0113862073324564241211064620.
Methotrexate (MTX) effectively eliminates cancerous cells but can also cause inflammation intestinal, known as mucositis. Forsythiaside A (FTA) from Forsythia suspensa has shown promise in relieving mucositis by targeting the NLRP3 pathways. Since NLRP3 inflammasome activation is negatively regulated by autophagy, this study explores how FTAmediated autophagy affects NLRP3 inflammasome in treating MTX-induced intestinal inflammation.
Intestinal mucositis was induced in rats with MTX. FTA's impact was assessed using HE staining and ELISA. The mechanism was studied using immunofluorescence, western blot, and ELISA.
FTA treatment resulted in reduced levels of D-lactic acid and diamine oxidase (DAO) in MTX-treated rats. Western blot and immunofluorescence analyses revealed up-regulation of Beclin- 1 and LC3II/I, accumulation of LC3, and down-regulation of p62 expression levels in MTXtreated rats following 40 or 80 mg/kg FTA intervention. However, when the autophagy inhibitor 3-MA was used, the intestinal pathology was exacerbated, the inflammatory scores increased, and serum levels of TNF-α, IL-1β, and IL-18 were elevated. Western blotting indicated decreased LC3II/I expression, while NLRP3, cleaved caspase 1, and cleaved IL-1β expressions were upregulated.
These findings suggested that FTA alleviated MTX-treated intestinal mucositis by activating autophagy, which in turn inhibits the NLRP3 inflammasome.
甲氨蝶呤（MTX）可有效消除癌细胞，但也会引发肠道炎症，即黏膜炎。连翘中的连翘酯苷A（FTA）已显示出通过靶向NLRP3信号通路缓解黏膜炎的潜力。由于NLRP3炎性小体的激活受自噬负调控，本研究探讨FTA介导的自噬在治疗MTX诱导的肠道炎症中如何影响NLRP3炎性小体。
用MTX诱导大鼠肠道黏膜炎。通过苏木精-伊红（HE）染色和酶联免疫吸附测定（ELISA）评估FTA的作用。使用免疫荧光、蛋白质免疫印迹法和ELISA研究其机制。
FTA治疗使MTX处理的大鼠体内D-乳酸和二胺氧化酶（DAO）水平降低。蛋白质免疫印迹法和免疫荧光分析显示，在40或80mg/kg FTA干预后，MTX处理的大鼠体内Beclin-1和LC3II/I上调，LC3积累，p62表达水平下调。然而，当使用自噬抑制剂3-甲基腺嘌呤（3-MA）时，肠道病理状况恶化，炎症评分增加，血清肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18）水平升高。蛋白质免疫印迹法表明LC3II/I表达降低，而NLRP3、裂解的半胱天冬酶-1和裂解的IL-1β表达上调。
这些发现表明，FTA通过激活自噬减轻MTX诱导的肠道黏膜炎，进而抑制NLRP3炎性小体。
