Objective of the study: We used fluorescence imaging methods of apoptosis and necrosis in human renal carcinoma A498 tumor cells in vitro to reveal the indicated forms of cell death under the combined effect of flavonoid-containing extract of Gratiola officinalis and cytostatic (cyclophosphamide). Materials and methods: The dyes were propidium iodide and acridine orange, which were used in the “alive and dead” test. This test helped us to identify the total number of dead cells in the forms of necrosis and apoptosis and the number of cells in which apoptosis had started, it was characterized by the appearance of apoptotic bodies or nucleus pyknosis. Results: We found the most pronounced cytotoxic activity at the ratio of extract of Gratiola officinalis and cyclophosphamide concentrations of 1:1. The number of living cells decreased when exposed to the ratio of extract and cytostatic concentrations of 2:1. When the ratio of concentration of the extract relative to the cytostatic increased to 3:1, the cytostatic activity of the extract began to appear, the total number of tumor cells decreased. The number of cells with nucleus pyknosis and the number of cells with apoptosis signs significantly increased at a 3:1 ratio of extract and cytostatic concentrations, which confirms the presence of pro-apoptotic activity of the studied combination. This trend indicates the dependence of a certain form of cell death (apoptosis, necrosis) on the ratio of extract and cytostatic doses, and it also demonstrates the cytostatic and cytotoxic effects of this combination. Conclusion: Fluorescence methods of investigation in the “alive and dead” test allowed us to visualize the forms of cell death of human kidney carcinoma A498 by combined exposure to the flavonoid-containing extract of Gratiola officinalis and cytostatic (cyclophosphamide) 24 h after exposure. We found that the combination with a concentration ratio of the extract and cyclophosphamide of 3:1 has the greatest effectiveness due to stimulation of the cytostatic effect and cytotoxic effect.

中文翻译：

---

使用荧光染色法在葛兰素提取物和环磷酰胺联合作用下肾细胞癌的肿瘤细胞死亡可视化

研究目的: 我们使用荧光成像方法检测人肾癌 A498 肿瘤细胞的凋亡和坏死体外在含有黄酮类化合物的提取物的联合作用下揭示细胞死亡的指示形式香菜和细胞抑制剂（环磷酰胺）。材料和方法：染料是碘化丙啶和吖啶橙，用于“生死”测试。该测试帮助我们确定坏死和凋亡形式的死细胞总数以及开始凋亡的细胞数量，其特征在于出现凋亡小体或核固缩。结果：我们发现在提取物的比例下最显着的细胞毒活性香菜和环磷酰胺浓度为1:1。当提取物和细胞抑制剂浓度为 2:1 时，活细胞的数量减少。当提取物相对于细胞抑制剂的浓度比增加到3:1时，提取物的细胞抑制活性开始显现，肿瘤细胞总数减少。在提取物和细胞抑制浓度的比例为 3:1 时，具有核固缩的细胞数量和具有凋亡迹象的细胞数量显着增加，这证实了所研究组合的促凋亡活性的存在。这一趋势表明某种形式的细胞死亡（细胞凋亡、坏死）对提取物和细胞抑制剂剂量的比例的依赖性，并且它也证明了这种组合的细胞抑制和细胞毒性作用。结论：“活与死”测试中的荧光研究方法使我们能够通过联合暴露于含黄酮类化合物的提取物来观察人肾癌 A498 的细胞死亡形式。香菜和细胞抑制剂（环磷酰胺）暴露后 24 小时。我们发现提取物和环磷酰胺的浓度比为 3:1 的组合具有最大的效果，因为它刺激了细胞抑制作用和细胞毒作用。