To systematically compare the cell damage characteristics of stretch injury model, oxygen-glucose deprivation (OGD) model and hydrogen peroxide (H₂O₂)-treated oxidative stress model, and to provide a reference for the selection of cell models in TBI research.

After resuscitation of frozen HT-22 cells, HT-22 cells were seeded in Six-well plates, which were divided into experimental group and control group. The experimental group prepared cell stretch injury models, OGD models, and oxidative stress injury models respectively. The control group was cultured normally without any special treatment. The changes of cell morphology were observed by microscope, and cell viability was tested by CCK-8 analysis; cell proteins were extracted, and changes of apoptosis markers (Bcl-2, Bax), necroptosis markers (RIPK3, RIPK1), ferroptosis markers (ACSL4, GPX4, SLC7A11) were detected by Western blot.

Compared with the control group, all three injury models led to abnormal morphological changes in different degrees, restricted proliferation or cell shedding in HT-22 cells, indicating cell damage. CCK-8 analysis showed that the cell viability in different model treatment groups was significantly reduced (P<0.05). The results of Western blot demonstrated that the expression of GPX4, SLC7A11, Bax, RIPK3 and RIPK1 proteins in cells treated with 3.0 PSI tensile strength was significantly increased, with statistical significance (P<0.05). After OGD treatment for 12 h, the expression of SLC7A11 and Bax was significantly up-regulated, with statistical significance (P<0.05). After 800 μmol/L hydrogen peroxide treatment, the expression of Bcl-2 and RIPK1 proteins was significantly up-regulated, with statistical significance (P<0.05).

The stretch model can better reflect the complex injury characteristics after TBI, and the OGD and H₂O₂ treatment models are more suitable for studying specific injury pathways.