KGF 联合 HIF-1琢 对 BECN1 敲除 IEC-6 细胞缺氧的保护作用及机制.

Objective: To investigate the protective effect and mechanism of KGF combined with HIF-1α on IEC-6 cells knocked out by BECN1 under hypoxia stress. Methods: The stable BECN1 knockout cell line IEC-6B-cells were constructed and divided into blank control group (BC), negative control group (NC) and HIF-1 α Group, KGF joint HIF-1 α Group (KH). Observed the changes of cell morphology in each group. Cell survival rate, ATP content, cell cycle and apoptosis rate, autophagy related gene expression, apoptosis and autophagy related protein expression were detected in each group. Results: After 24 hours of hypoxia treatment, spindle, star and other abnormal morphological changes were observed in cells of each group. Electron microscopy showed that a large number of lysosomal vesicles with different sizes were found in the cytoplasm of NC cells. Other groups of cells showed morphological changes of early apoptosis, and autophagic vesicles were found in the cytoplasm of cells in BC group and KH group. Compared with BC group, the cell survival rate of each knockout group was significantly lower (P＜0.01), and the cell survival rate of KH group was higher than NC group,KGF group and HIF-1α group (P＜0.05). The content of ATP in BC group was significantly higher than that in other groups (P＜0.05).Compared with BC group and KH group, the percentage of G0/G1 phase and apoptosis rate of other three groups were significantly increased (P＜0.05). There was no significant difference between KH group and BC group (P＞0.05). Compared with BC group, the expression of BECN1, SQSTM1 and LC3 genes in NC group, KGF group, HIF-1α group and KH group decreased significantly (P＜0.05). Compared with BC group, the expression of Beclin1 protein and LC3 II/LC3 I in NC group, KGF group, HIF-1 α group and KH group decreased significantly (P＜0.05), and the expression of p62 protein increased significantly (P＜0.05). The ratio of Bax/Bcl-2 and the expression of Caspase3 protein in KH group were lower than those in NC group (P＜0.05). Conclusion: KGF combined with HIF-1α can protect IEC-6B-cells from hypoxia stress by promoting cell proliferation, enhancing cell energy metabolism, reducing cell arrest rate in G0/G1phase and inhibiting apoptosis. [ABSTRACT FROM AUTHOR]