Ajmalicine (AJM) is an alkaloid extracted from the root of Yunan Rauvolfia verticillata. At present, little research has reported the antitumor pharmacological action and mechanism of AJM. Therefore, this work aimed to conduct relevant research. The mouse hepatoma cell line H22 was intervened with a gradient concentration of AJM. Subsequently, the pyroptosis level was detected by flow cytometry. The expression of inflammatory factors and lactate dehydrogenase was measured by enzyme-linked immunosorbent assay. Reactive oxygen species (ROS) expression was detected by dichlorodihydrofluorescein diacetate probe. In addition, the tumor-bearing model mice were also treated with AJM to analyze tumor growth as well as the expression levels of tissue inflammatory factors and proteins. According to our results, AJM promoted the pyroptosis of H22 cells, increased the pyroptosis rate, and upregulated the expression of inflammatory factors tumor necrosis factor α, interleukin-1β, and interleukin-6. At the same time, it enhanced the openness of membrane pores and increased the expression of ROS. Moreover, AJM promoted the expression of Caspase-3 and N-terminal gasdermin E (GSDME). The AJM-induced pyroptosis was suppressed after N-acetylcysteine treatment to inhibit ROS, while Caspase-3 knockdown also inhibited the AJM-induced pyroptosis. In animals, AJM suppressed tumor growth. AJM can activate ROS to induce pyroptosis and exert the antitumor effect via the noncanonical Caspase-3-GSDME pyroptosis pathway.

中文翻译：

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Ajmalicine诱导肝癌细胞焦亡发挥抗肿瘤作用


Ajmalicine (AJM) 是从云南萝芙木根中提取的生物碱。目前，AJM抗肿瘤药理作用及机制的研究报道较少。因此，本工作旨在开展相关研究。用梯度浓度的AJM对小鼠肝癌细胞系H22进行干预。随后，通过流式细胞术检测细胞焦亡水平。采用酶联免疫吸附法测定炎症因子和乳酸脱氢酶的表达。通过二氯二氢荧光素二乙酸酯探针检测活性氧（ROS）表达。此外，还用AJM治疗荷瘤模型小鼠，分析肿瘤生长以及组织炎症因子和蛋白质的表达水平。根据我们的结果，AJM促进H22细胞焦亡，增加焦亡率，并上调炎症因子肿瘤坏死因子α、白细胞介素1β和白细胞介素6的表达。同时增强膜孔的开放性，增加ROS的表达。此外，AJM 还能促进 Caspase-3 和 N 末端gasdermin E (GSDME) 的表达。 N-乙酰半胱氨酸处理抑制ROS后，AJM诱导的焦亡被抑制，而Caspase-3敲低也抑制AJM诱导的焦亡。在动物中，AJM 抑制肿瘤生长。 AJM 可以激活 ROS 诱导细胞焦亡，并通过非经典的 Caspase-3-GSDME 焦亡途径发挥抗肿瘤作用。