METHODS: In the research, the retinal Müller cells are regarded as research subjects, and the control group, hypoxia group and experiment group are set up. The cells of control group are cultivated in normal environment. The cells of hypoxia group are cultivated in hypoxia environment. The cells of experiment group are cultivated in hypoxia environment and are treated with the Apelin-13(1μmol/L). MTT method is used to monitor the changing of the cell viability, and the crystal violet staining method is adopted to observe the cell morphology. In addition, the immunofluorescence staining method is used to test the expression of GFAP and YAP and the TUNEL staining method is used to monitor the cell apoptosis situation and the apoptosis index is calculated. The protein staining method is used to observe the changing of the expression of p-LATS1, p-YAP, LATS1 and YAP protein.

RESULTS:The separated and extracted Müller cells grow on the wall and show elongation, polygon and circular shapes. The cytoplasm is plentiful and the cell nucleus show circular shape. The GFAP expression of the cell is positive. The treatment with 0.1, 1, 10μmol/L Apelin-13 can obviously prevent the Müller cell viability decreasing induced by hypoxia(P<0.05 or P<0.01). Compared with the control group, the cell apoptosis index of hypoxia group is obviously increased(P<0.01). However, compared with the hypoxia group, the cell apoptosis index of experiment group is obviously decreased(P<0.01). The p-LATS1 and p-YAP protein expression of the control group and hypoxia group does not have big difference. Compared with hypoxia group, the p-LATS1 and p-YAP protein expression of experiment group is obviously decreased(P<0.01). The YAP protein expression of cell nucleus of control group and hypoxia group does not have great difference. Compared with hypoxia group, the cell nucleus expression of YAP cell is gretaly increased(P<0.01).

CONCLUSION: Apelin-13 can be used to prevent the retinal Müller cells apoptosis caused by the hypoxia, which may be related to the regulation of YAP into the nucleus.