Department of Nephrology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Research Institute of Nephrology, Zhengzhou University, Zhengzhou, China.
Front Immunol. 2024 Sep 25;15:1457230. doi: 10.3389/fimmu.2024.1457230. eCollection 2024.
Cisplatin is a widely used chemotherapeutic agent prescribed to treat solid tumors. However, its clinical application is limited because of cisplatin- induced nephrotoxicity. A known complication of cisplatin is acute kidney injury (AKI). Deletion polymorphisms of GSTM1 and GSTT1, members of the glutathione S-transferase family, are common in humans and are presumed to be associated with various kidney diseases. However, the specific roles and mechanisms of GSTM1 and GSTT1 in cisplatin induced AKI remain unclear.
To investigate the roles of GSTM1 and GSTT1 in cisplatin-induced AKI, we generated GSTM1 and GSTT1 knockout mice using CRISPR-Cas9 technology and assessed their kidney function under normal physiological conditions and cisplatin treatment. Using ELISA kits, we measured the levels of oxidative DNA and protein damage, along with MDA, SOD, GSH, and the GSH/GSSG ratio in wild-type and GSTM1/GSTT1 knockout mice following cisplatin treatment. Additionally, oxidative stress levels and the expression of ferroptosis-related proteins in kidney tissues were examined through Western blotting, qPCR, immunohistochemistry, and immunofluorescence techniques.
Here, we found that GSTT1 and GSTM1 were downregulated in the renal tubular cells of AKI patients and cisplatin-treated mice. Compared with WT mice, -DKO mice were phenotypically normal but developed more severe kidney dysfunction and exhibited increased ROS levels and severe ferroptosis after injecting cisplatin.
Our study revealed that GSTM1 and GSTT1 can protect renal tubular cells against cisplatin-induced nephrotoxicity and ferroptosis, and genetic screening for GSTM1 and GSTT1 polymorphisms can help determine a standard cisplatin dose for cancer patients undergoing chemotherapy.
顺铂是一种广泛应用于治疗实体瘤的化疗药物。然而，由于顺铂诱导的肾毒性，其临床应用受到限制。顺铂的已知并发症之一是急性肾损伤（AKI）。谷胱甘肽 S-转移酶家族成员 GSTM1 和 GSTT1 的缺失多态性在人类中很常见，据推测与各种肾脏疾病有关。然而，GSTM1 和 GSTT1 在顺铂诱导的 AKI 中的具体作用和机制尚不清楚。
为了研究 GSTM1 和 GSTT1 在顺铂诱导的 AKI 中的作用，我们使用 CRISPR-Cas9 技术生成 GSTM1 和 GSTT1 敲除小鼠，并在正常生理条件和顺铂处理下评估其肾功能。使用 ELISA 试剂盒，我们测量了野生型和 GSTM1/GSTT1 敲除小鼠在顺铂处理后 DNA 和蛋白质氧化损伤的水平，以及 MDA、SOD、GSH 和 GSH/GSSG 比值。此外，通过 Western blot、qPCR、免疫组织化学和免疫荧光技术检测了肾组织中的氧化应激水平和铁死亡相关蛋白的表达。
在这里，我们发现 AKI 患者和顺铂处理小鼠的肾小管细胞中 GSTT1 和 GSTM1 下调。与 WT 小鼠相比，-DKO 小鼠表型正常，但在注射顺铂后肾脏功能障碍更严重，ROS 水平升高，铁死亡更严重。
我们的研究表明，GSTM1 和 GSTT1 可以保护肾小管细胞免受顺铂诱导的肾毒性和铁死亡，对 GSTM1 和 GSTT1 多态性进行基因筛查有助于确定接受化疗的癌症患者的标准顺铂剂量。
