Department of Critical Care Medicine, Union Jiangbei Hospital, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.
J Leukoc Biol. 2022 Nov;112(5):1065-1077. doi: 10.1002/JLB.1A0422-211R. Epub 2022 Jun 30.
Acute kidney injury (AKI) represents a prevailing complication of sepsis, and its onset involves ferroptosis. Ginsenoside Rg1 exerts a positive effect on kidney diseases. This study explored the action of ginsenoside Rg1 in sepsis-induced AKI (SI-AKI) by regulating ferroptosis in renal tubular epithelial cells (TECs). Sepsis rat models were established using cecal ligation and puncture (CLP) and cell models were established by treating human renal TECs (HK-2) with LPS to induce ferroptosis. Serum creatinine (SCr) and blood urea nitrogen (BUN) and urine KIM1 contents in rats were determined by ELISA kits. Kidney tissues were subjected to immunohistochemical and H&E stainings. Iron concentration, malondialdehyde (MDA), glutathione (GSH), and ferroptosis-related protein (ferritin light chain [FTL], ferritin heavy chain [FTH], GSH peroxidase 4 [GPX4], and Ferroptosis suppressor protein 1 [FSP1]) levels in kidney tissues and HK-2 cells were measured using ELISA kits and Western blotting. HK-2 cell viability was detected by cell counting kit-8, and cell death was observed via propidium iodide staining. Reactive oxygen species accumulation in cells was detected using C11 BODIPY 581/591 as a molecular probe. In CLP rats, ginsenoside Rg1 reduced SCr, BUN, KIM1, and NGAL levels, thus palliating SI-AKI. Additionally, ginsenoside Rg1 decreased iron content, FTL, FTH, and MDA levels, and elevated GPX4, FSP1, and GSH levels, thereby inhibiting lipid peroxidation and ferroptosis. Moreover, FSP1 knockdown annulled the inhibition of ginsenoside Rg1 on ferroptosis. In vitro experiments, ginsenoside Rg1 raised HK-2 cell viability and lowered iron accumulation and lipid peroxidation during ferroptosis, and its antiferroptosis activity was dependent on FSP1. Ginsenoside Rg1 alleviates SI-AKI, possibly resulting from inhibition of ferroptosis in renal TECs through FSP1.
急性肾损伤（AKI）是脓毒症的主要并发症之一，其发生涉及铁死亡。人参皂苷 Rg1 对肾脏疾病有积极作用。本研究通过调节肾小管上皮细胞（TEC）中的铁死亡来探讨人参皂苷 Rg1 在脓毒症诱导的急性肾损伤（SI-AKI）中的作用。采用盲肠结扎穿孔（CLP）建立脓毒症大鼠模型，并用 LPS 处理人肾 TEC（HK-2）细胞建立铁死亡细胞模型。通过 ELISA 试剂盒测定大鼠血清肌酐（SCr）和血尿素氮（BUN）及尿 KIM1 含量。对肾脏组织进行免疫组化和 H&E 染色。采用 ELISA 试剂盒和 Western blot 法测定肾组织和 HK-2 细胞中铁浓度、丙二醛（MDA）、谷胱甘肽（GSH）和铁死亡相关蛋白（铁蛋白轻链[FTL]、铁蛋白重链[FTH]、谷胱甘肽过氧化物酶 4[GPX4]和铁死亡抑制蛋白 1[FSP1]）水平。通过细胞计数试剂盒-8 检测 HK-2 细胞活力，通过碘化丙啶染色观察细胞死亡。采用 C11 BODIPY 581/591 作为分子探针检测细胞内活性氧（ROS）积累。在 CLP 大鼠中，人参皂苷 Rg1 降低了 SCr、BUN、KIM1 和 NGAL 水平，从而缓解了 SI-AKI。此外，人参皂苷 Rg1 降低了铁含量、FTL、FTH 和 MDA 水平，提高了 GPX4、FSP1 和 GSH 水平，从而抑制了脂质过氧化和铁死亡。此外，FSP1 敲低消除了人参皂苷 Rg1 对铁死亡的抑制作用。在体外实验中，人参皂苷 Rg1 提高了 HK-2 细胞活力，降低了铁积累和铁死亡期间的脂质过氧化，其抗铁死亡活性依赖于 FSP1。人参皂苷 Rg1 缓解 SI-AKI，可能是通过 FSP1 抑制肾 TEC 中的铁死亡。
