Eupatilin alleviated ferroptosis and epithelial-mesenchymal transition in high glucose-induced HK2 cells by regulating Nrf2 pathway
10.3760/cma.j.cn121382-20250203-00014
- VernacularTitle:泽兰林素通过调控Nrf2通路缓解高糖诱导的HK2细胞铁死亡和上皮-间充质转化
- Author:
Chunxiao XIE
1
;
Penghao LI
;
Li WANG
Author Information
1. 天津医科大学第二医院健康管理科，天津 300211
- Keywords:
Diabetic nephropathy;
Eupatilin;
High glucose;
Ferroptosis;
Epithelial-mesenchymal transition;
Nuclear factor-erythroid 2-related factor 2
- From:
International Journal of Biomedical Engineering
2025;48(3):232-238
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of eupatilin on ferroptosis and epithelial-mesenchymal transition (EMT) in high glucose-induced human renal tubular epithelial HK2 cells and its mechanism.Methods:HK2 cells were divided into a blank control group and a high glucose group according to the glucose concentration in the medium. The cells were cultured in RPMI 1640 medium containing 5.6 or 30.0 mmol/L glucose. Additionally, an eupatilin control group and an eupatilin group were established, and 1 μmol/L eupatilin was added to the blank control group and the high glucose group, respectively. Cell proliferation and apoptosis rates were evaluated using a cell counting kit-8 assay and a flow cytometry assay, respectively. The relative reactive oxygen species content was detected using 2′,7′-dichlorodihydrofluorescein diacetate fluorescent probe method and the relative expression of glutathione peroxidase 4 (GPX4) protein was detected using Western blotting to investigate eupatilin′s regulatory effect on ferroptosis. The effect of eupatilin on mitochondrial damage was detected by JC-1 staining. The relative expression of the EMT marker vimentin and nuclear factor-erythroid 2-related factor 2 (Nrf2) protein were evaluated using Western blotting. Data were analyzed by an independent sample t test or one-way analysis of variance. Results:Eupatilin treatment promoted the viability of HK2 cells induced by high glucose. The cell proliferation rate in the eupatilin group [(92.00±6.00)%] was higher than that in the high glucose group [(70.00±4.00)%], and the difference was statistically significant ( t=5.284, P<0.01). Eupatilin treatment inhibited the apoptosis of HK2 cells induced by high glucose. The apoptosis rate of HK2 cells in the eupatilin group [(5.00±1.50)%] was lower than that in the high glucose group [(43.00±4.00)%], and the difference was statistically significant ( t=15.410, P<0.01). Eupatilin blocked the ferroptosis in HK2 cells induced by high glucose. The relative reactive oxygen species content in the eupatilin group (1.50±0.23) was lower than that in the high glucose group (3.20±0.21), and the difference was statistically significant ( t=9.454, P<0.01). The relative expression of GPX4 protein in the eupatilin group (0.89±0.20) was higher than that in the high glucose group (0.21±0.02), and the difference was statistically significant ( t=6.721, P<0.01). Eupatilin reduced the mitochondrial damage in HK2 cells induced by high glucose. The red-green fluorescence intensity ratio in the eupatilin group (0.65±0.12) was higher than that in the high glucose group (0.32±0.11), and the difference was statistically significant ( t=3.298, P<0.01). Eupatilin inhibited the EMT process in HK2 cells induced by high glucose. The relative expression of vimentin in the eupatilin group (1.32±0.20) was lower than that in the high glucose group (2.12±0.12), and the difference was statistically significant ( t=5.941, P<0.01). Eupatilin activated the Nrf2 pathway in HK2 cells induced by high glucose. The relative expression of Nrf2 protein in the eupatilin group (0.87±0.12) was higher than that in the high glucose group (0.45±0.14), and the difference was statistically significant ( t=3.945, P<0.01). Conclusions:Eupatilin may inhibit ferroptosis and EMT process in high glucose-induced HK2 cells by activating the Nrf2 pathway.
