Effect of Qijudihuang Pill-Containing Serum on the Process of MitochondrialApoptosis in Corneal Epithelial Cells Induced by Hyperosmolarity

This study aims to evaluate the protective effect of Qijudihuang pill-containing serum on hyperosmolarity-induced HCE-T corneal epithelial cell injury, and to explore its relationship with mitochondrial apoptosis-relatedprocesses. HCE-T cells were cultured in vitro, and the final osmotic pressure of the medium was adjusted to 500 mOsmol/Lin DMEM/F12 culture system to establish a dry eye related cell injury model induced by hyperosmolarity in vitro. The cells were treated with different volume fractions of Qijudihuang pill-containing serum. After screening the optimal interventionconditions, the cells were divided into Control, Model and QJDH groups. Cell viability was detected by CCK-8 assay. Thelevels of IL-1β, IL-6, IL-17 and TNF-α in the culture supernatant were detected by ELISA. The levels of MDA (malondialdehyde) and SOD (superoxide dismutase) in the cells and LDH in the supernatant were detected by kits. The activity of Caspase-9 was detected by fluorescent microplate reader. Annexin V-FITC/PI flow cytometry was used to detect the apoptosisrate, and TF2-DEVD-FMK flow cytometry was used to detect the activation of Caspase-3/7. DCFH-DA, TMRE and MitoSOXTM were used to detect the levels of ROS, mitochondrial membrane potential and mitochondrial superoxide, respectively. The protein levels of Bax, Bcl-2, AIF, EndoG and Cyt-C were detected by Western blot. Compared with the Controlgroup, the cell viability and SOD level were decreased, the inflammatory factor (IL-1β, IL-6, IL-17, TNF-α) levels, MDAlevel, LDH level, apoptosis rate, Caspase-9 activity, Caspase-3/7 activation level, ROS level and mitochondrial superoxidelevel were increased, and mitochondrial membrane potential was decreased in the Model group. The protein levels of Bax,AIF, EndoG and Cyt-C were increased, and the level of Bcl-2 was decreased (P<0.05). Compared with the Model group,the above indicators in the QJDH group showed opposite changes (P<0.05). Qijudihuang pill-containing serum can alleviate hyperosmolarity-induced HCE-T corneal epithelial cell injury, accompanied by the reduction of oxidative stress, theimprovement of mitochondrial function and the down-regulation of mitochondrial apoptosis related indicators, suggestingthat its protective effect may be related to the regulation of mitochondrial apoptosis related processes, providing a potentialexperimental basis for clinical intervention of corneal epithelial cell injury related diseases in hyperosmolarity environment.It also lays a research foundation for further exploring the target and specific molecular mechanism of Qijudihuang pill.

CSTR: 32200.14.cjcb.2026.10.0005