Chu Ying-Ming, Wang Ting-Xin, Jia Xiao-Fen, Yang Yao, Shi Zong-Ming, Cui Guang-Hui, Huang Qiu-Yue, Ye Hui, Zhang Xue-Zhi
Department of Integrated Traditional Chinese and Western Medicine, Peking University First Hospital, Institute of Integrated Traditional Chinese and Western Medicine, Peking University, Beijing, China.
Institute (College) of Integrative Medicine, Dalian Medical University, Dalian, China.
Front Pharmacol. 2022 Nov 23;13:1066244. doi: 10.3389/fphar.2022.1066244. eCollection 2022.
Fuzheng Nizeng Decoction (FZNZ) has a history of decades in gastric precancerous lesions (GPL) treatment, which has shown clear clinical efficacy. Blocking GPL is a key measure to reduce the incidence of gastric cancer (GC). Therefore, we aim to investigate the mechanism of FZNZ-induced ferroptosis and endoplasmic reticulum (ER) in MNNG-induced gastric precancerous lesion (MC) cells, which has been rarely studied in Traditional Chinese Medicine (TCM). First, CCK8 and lactate dehydrogenase assays were conducted to study the potential effect of FZNZ on MC cells. Second, combined transcriptomic and metabolomic analysis were used to explore the effect and mechanism of FZNZ. Functionally, the occurrence of ferroptosis was assessed by transmission electron microscopy morphological observation and measurement of ferrous iron levels, lipid peroxidation, and glutathione levels. Finally, the expression levels of mRNAs or proteins related to ferroptosis and ER stress were determined by qPCR or western blot assays, respectively. FZNZ inhibited MC cells viability and induced cell death. By metabolomics coupled with transcriptomics analysis, we found that the mechanism of FZNZ treatment induced ferroptosis and was related to glutathione metabolism and ER stress. We then, for the first time, found that FZNZ induced ferroptosis, which contributed to an increase in intracellular ferrous iron, reactive oxygen species, and malondialdehyde and a decrease in glutathione. Meanwhile, the protein level of glutathione peroxidase 4 (GPX4) was decreased. The mRNA levels of ATF3/CHOP/CHAC1, which are related to ferroptosis and ER stress, were also upregulated. Our results elaborate that FZNZ could induce ferroptosis and ER stress in MC cells, and reduce GPX4/GSH. ATF3/CHOP/CHAC1 may play a crosstalk role, which provides a new molecular mechanism for the treatment of GPL.
扶正抑增方（FZNZ）在治疗胃癌前病变（GPL）方面已有数十年历史，已显示出明确的临床疗效。阻断GPL是降低胃癌（GC）发病率的关键措施。因此，我们旨在研究扶正抑增方诱导MNNG诱导的胃癌前病变（MC）细胞铁死亡和内质网（ER）应激的机制，而这在传统中医（TCM）中鲜有研究。首先，进行CCK8和乳酸脱氢酶检测以研究扶正抑增方对MC细胞的潜在作用。其次，采用转录组学和代谢组学联合分析来探索扶正抑增方的作用及机制。在功能上，通过透射电子显微镜形态学观察以及亚铁离子水平、脂质过氧化和谷胱甘肽水平的测定来评估铁死亡的发生情况。最后，分别通过qPCR或蛋白质免疫印迹法检测与铁死亡和内质网应激相关的mRNA或蛋白质的表达水平。扶正抑增方抑制MC细胞活力并诱导细胞死亡。通过代谢组学与转录组学联合分析，我们发现扶正抑增方治疗诱导铁死亡的机制与谷胱甘肽代谢和内质网应激有关。然后，我们首次发现扶正抑增方诱导铁死亡，这导致细胞内亚铁离子、活性氧和丙二醛增加，谷胱甘肽减少。同时，谷胱甘肽过氧化物酶4（GPX4）的蛋白质水平降低。与铁死亡和内质网应激相关的ATF3/CHOP/CHAC1的mRNA水平也上调。我们的结果表明，扶正抑增方可诱导MC细胞发生铁死亡和内质网应激，并降低GPX4/谷胱甘肽水平。ATF3/CHOP/CHAC1可能发挥相互作用，这为GPL的治疗提供了一种新的分子机制。
