Effect of α-Klotho on macrophage-vascular endothelial cell crosstalk in diabetic oxidative stress environment / 国际眼科杂志(Guoji Yanke Zazhi)
International Eye Science
; (12): 1020-1026, 2024.
Article
in Zh
| WPRIM
| ID: wpr-1032341
Responsible library:
WPRO
ABSTRACT
AIM:
To investigate the effects of overexpressing α-Klotho(KL)in RAW264.7 cells stimulated by oxidative stress on the proliferation, migration, tube-formation and tight junction of human umbilical vein endothelial cells(HUVECs).METHODS:
RAW264.7 cells were categorized into control, 4-hydroxynonenal(4HNE), and 4HNE+KL groups, with F4/80 expression assessed via immunofluorescence staining. Three groups of conditional media were prepared for HUVECs and culture divided into Mø-NC, Mø-4HNE, and Mø-4HNE+KL groups. Cell proliferation was evaluated using CCK8 assay, while scratch test and Transwell assays were employed to measure cell migration. Additionally, tube-formation assay was conducted to assess cell tubule formation, and Western blot assay was utilized to detect the protein expression levels of Claudin 5, Occludin and ZO 1.RESULTS:
The results of immunofluorescence staining showed that the fluorescence intensity of F4/80 of RAW264.7 cells in the 4HNE group was significantly enhanced compared with the control group, while that of F4/80 in the 4HNE+KL group was significantly decreased compared with the 4HNE group(all P<0.05). The CCK8 assay results revealed a significant increase in the proliferation of HUVECs in the M&#xF8;-4HNE group compared with the M&#xF8;-NC group. Conversely, the proliferation of the M&#xF8;-4HNE+KL group exhibited a significant decrease compared with that in the M&#xF8;-4HNE group(all P<0.01). The results of scratch test and Transwell assays demonstrated a significant increase in the migration of HUVECs in the M&#xF8;-4HNE group compared with the M&#xF8;-NC group, while the migration of the M&#xF8;-4HNE+KL group exhibited a significant decrease compared with the M&#xF8;-4HNE group(all P<0.01). In the tube-formation assay, it was observed that the number of tubes formed by HUVECs in the M&#xF8;-4HNE group was significantly increased compared with the M&#xF8;-NC group, while that of tubes formed in the M&#xF8;-4HNE+KL group was significantly decreased compared with the M&#xF8;-4HNE group(all P<0.01). Additionally, the Western blot results revealed a significant decrease in the relative expression levels of Claudin 5, Occludin, and ZO 1 in the M&#xF8;-4HNE group compared with the M&#xF8;-NC group. Conversely, in the M&#xF8;-4HNE+KL group, there was a significant increase in the relative expression levels of Claudin 5, Occludin, and ZO 1 compared to the M&#xF8;-4HNE group(all P<0.01).CONCLUSIONS:
KL inhibits the proliferation, migration, and tube-formation of HUVECs while enhancing the tight junction by changing the activation state of macrophages in the diabetic oxidative stress environment.
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Index:
WPRIM
Language:
Zh
Journal:
International Eye Science
Year:
2024
Type:
Article