Department of Nephrology, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou City, Guangdong Province, China.
Nephrotic Diagnosis And Treatment Center, The Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan City, Shandong Province, China.
Bioengineered. 2022 Mar;13(3):6627-6637. doi: 10.1080/21655979.2022.2045834.
Diabetic nephropathy (DN) is associated with inflammation. Platycodin D (PD) demonstrates anti-inflammatory activity. However, whether PD affects DN remains to be explored. Here, we aimed to discuss the role of PD in DN and its underlying mechanisms. High glucose (HG)-induced HK-2 cells were treated with PD, and cell viability was assessed using the Thiazolyl Blue Tetrazolium Bromide (MTT) assay. Ferroptosis-related factors such as lactate dehydrogenase (LDH) activity, lipid reactive oxygen species (ROS), iron (Fe) level, GSH level, and malondialdehyde (MDA) level were evaluated. Cell death was evaluated using the TUNEL assay. GPX4 expression was evaluated using Quantitative Real-time PCR (qRT-PCR) and Western blotting analysis. The results indicated that HG increased LDH activity, lipid ROS production, Fe levels, and MDA levels and decreased GSH levels, suggesting that the HG condition induced ferroptosis. PD treatment inhibited ferroptosis in HG-induced cells, downregulated ACSL4 and TFR1 expression, and upregulated FTH-1 and SLC7A11 expression. PD reversed the effects of HG condition on cell death. Moreover, GPX4 expression was downregulated in HG-stimulated cells. Furthermore, we substantiated that PD suppressed ferroptosis by modulating GPX4 expression. In conclusion, PD inhibited ferroptosis in HG-induced HK-2 cells by upregulating GPX4 expression, suggesting that PD may be an effective drug for the clinical treatment of DN.
糖尿病肾病（DN）与炎症有关。桔梗皂苷 D（PD）具有抗炎活性。然而，PD 是否影响 DN 仍有待探讨。在这里，我们旨在讨论 PD 在 DN 中的作用及其潜在机制。用 PD 处理高葡萄糖（HG）诱导的 HK-2 细胞，使用噻唑蓝四唑溴盐（MTT）测定法评估细胞活力。评估铁死亡相关因素，如乳酸脱氢酶（LDH）活性、脂质活性氧（ROS）、铁（Fe）水平、GSH 水平和丙二醛（MDA）水平。使用 TUNEL 测定法评估细胞死亡。使用定量实时 PCR（qRT-PCR）和 Western blot 分析评估 GPX4 表达。结果表明，HG 增加了 LDH 活性、脂质 ROS 产生、Fe 水平和 MDA 水平，降低了 GSH 水平，表明 HG 条件诱导了铁死亡。PD 处理抑制了 HG 诱导的细胞中铁死亡，下调 ACSL4 和 TFR1 表达，上调 FTH-1 和 SLC7A11 表达。PD 逆转了 HG 条件对细胞死亡的影响。此外，GPX4 在 HG 刺激的细胞中表达下调。此外，我们证实 PD 通过调节 GPX4 表达抑制铁死亡。总之，PD 通过上调 GPX4 表达抑制 HG 诱导的 HK-2 细胞中的铁死亡，表明 PD 可能是治疗 DN 的有效药物。
