Study on the protective mechanism of Qishen recipe on corneal epithelial cells induced by hypertonic based on JNK1/ AQP5 pathway / 国际眼科杂志(Guoji Yanke Zazhi)

ABSTRACT

@#AIM: To observe the protective effect of Qishen recipe on corneal epithelial cells induced by hypertonic fluid, and elucidated its mechanism of action in the treatment of dry eye base on JNK1 / AQP5 pathway.<p>METHODS: Human corneal epithelial cells(HCECs)model was created by osmotic pressure at a concentration of 500mOsm/L for 24h. Serum of rats containing drugs in the blank group, model group, Western medicine group, and Qishen recipe low-dose, medium-dose and high-dose groups were treated on the modeled DE HCECs, and the effects of different drug stimulation on the survival rate of HCECs were tested by CCK-8 method. The expressions of inflammatory factors TNF-α, IL-6 in extracellular fluid were explored by ELISA. The expression of apoptosis factors caspase 1 and AQP5 were detected by immunocytochemistry(ICC). The expressions of AQP5, JNK1, p-JNK1 of HCECs after intervention with different drug concentrations were found by Western blotting.<p>RESULTS: Compared with the blank group, the survival rate of HCECs in each group was significantly reduced(<i>P</i><0.01). The extracellular fluid inflammatory factors TNF-α, IL-6 and caspase-1, p-JNK1, AQP5 protein expression levels increased significantly in each group(all <i>P</i><0.01); In comparison to the model group, the survival rate of HCECs in each medication group increased significantly(all <i>P</i><0.01). The expression levels of TNF-α, IL-6 in the extracellular fluid of each drug group, AQP 5 and p-JNK1 protein expression in HCECs, and the expression of caspase-1 and AQP5 protein in the western medicine group and the Qishen recipe high and medium dose group were all reduced(all <i>P</i><0.05). Compared with the western medicine group, the survival rate of HCECs in the Qishen prescription high-dose group was significantly increased(<i>P</i><0.01). The expression levels of TNF-α and IL-6 in each dose group of Qishen recipe were reduced(all <i>P</i><0.05), while the expression levels of caspase-1 in the high-dose Qishen recipe group and the AQP5 protein expression levels of the high and medium-dose Qishen recipe group saw a decrease(all <i>P</i><0.05). However, there was no statistically significant difference in the JNK1 protein expression of HCECs of all the groups detected by Western blotting method(<i>P</i>>0.05). <p>CONCLUSION: Qishen recipe can not only reduce the JNK1 phosphorylation and AQP5 protein expression of HCECs induced by hypertonicity, but also reduce the expression of inflammatory factors TNF-α, IL-6 and the apoptotic factor caspase-1 of HCECs in the extracellular fluid, thus effectively Inhibit inflammation and apoptosis.