Effect of α-Klotho on macrophage-vascular endothelial cell crosstalk in diabetic oxidative stress environment
10.3980/j.issn.1672-5123.2024.7.03
- VernacularTitle:糖尿病氧化应激环境中α-Klotho对巨噬细胞-血管内皮细胞串扰的影响
- Author:
Qingbo LI
1
,
2
,
3
,
4
,
5
;
Peiyu WANG
1
,
2
,
3
,
4
,
5
;
Liying HU
1
,
2
,
3
,
4
,
5
;
Xiaorong LI
1
,
2
,
3
,
4
,
5
;
Yan SHAO
1
,
2
,
3
,
4
,
5
Author Information
1. Tianjin Medical University Eye Hospital
2. School of Optometry &
3. Eye Institute
4. Tianjin Key Laboratory of Retinal Functions and Diseases
5. Tianjin International Joint Research and Development Centre of Ophthalmology and Vision Science, Tianjin 300384, China
- Publication Type:Journal Article
- Keywords:
oxidative stress;
macrophage;
vascular endothelial cells;
proliferation;
migration;
tube-formation;
tight junction;
retinal neovascularization
- From:
International Eye Science
2024;24(7):1020-1026
- CountryChina
- Language:Chinese
-
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 Mø-4HNE group compared with the Mø-NC group. Conversely, the proliferation of the Mø-4HNE+KL group exhibited a significant decrease compared with that in the Mø-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 Mø-4HNE group compared with the Mø-NC group, while the migration of the Mø-4HNE+KL group exhibited a significant decrease compared with the Mø-4HNE group(all P<0.01). In the tube-formation assay, it was observed that the number of tubes formed by HUVECs in the Mø-4HNE group was significantly increased compared with the Mø-NC group, while that of tubes formed in the Mø-4HNE+KL group was significantly decreased compared with the Mø-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 Mø-4HNE group compared with the Mø-NC group. Conversely, in the Mø-4HNE+KL group, there was a significant increase in the relative expression levels of Claudin 5, Occludin, and ZO 1 compared to the Mø-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.
