Study on effect and mechanism of Zhenzhu Mingmu Eye Drops protected HCEC against hypermeability-induced injury / 中草药

ABSTRACT

Objective: To investigate the effects and mechanism of Zhenzhu Mingmu Eye Drops (ZMED) on human corneal epithelial cells (HCEC) injury model induced by high permeability. Methods: The dye eyes disease model was established using 110 mmol/L NaCl to stimulate HCEC, and different concentration of ZMED was given to the treatment group. MTT assay and lactate dehydrogenase (LDH) leakage were used to evaluate the protection effects of ZMED on HCEC. The effects of ZMED on apoptosis of HCEC were analyzed by flow cytometry. AO/EB double staining and Hoechst 33258 analysis were used as sensitive assays for apoptosis. Moreover, HCEC treated with ZMED were subjected to Western blotting for protein levels of Caspase-9, Caspase-3 and PARP. Results: ZMED significantly increased cell viability of HCEC and decreased the LDH release in a dose-dependent manner. ZMED dramatically decreased the apoptosis rate. Furthermore, ZMED down-regulated the levels of cleaved Caspase-9, cleaved Caspase-3 and cleaved PARP. Conclusion: ZMED could protect HCEC against hypermeability, which may be related to the inhibition of Caspase-9/Casepase-3 signaling pathways.