Effect of transferrin glycation induced by high glucose on HK-2 cells in vitro
Background and objectiveGlycation is a common post-transcriptional modification of proteins. Previous studies have shown that advanced glycation end modified transferrin (AGE-Tf) levels in diabetic rat kidney tissues were increased; however, its role in diabetic nephropathy remains unclear. In this...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-01-01
|
| Series: | Frontiers in Endocrinology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fendo.2022.1009507/full |
| _version_ | 1797942740242137088 |
|---|---|
| author | Yanqi Ma Qikai Zhou Pingping Zhao Xiaoyu Lv Caixia Gong Jie Gao Jingfang Liu Jingfang Liu |
| author_facet | Yanqi Ma Qikai Zhou Pingping Zhao Xiaoyu Lv Caixia Gong Jie Gao Jingfang Liu Jingfang Liu |
| author_sort | Yanqi Ma |
| collection | DOAJ |
| description | Background and objectiveGlycation is a common post-transcriptional modification of proteins. Previous studies have shown that advanced glycation end modified transferrin (AGE-Tf) levels in diabetic rat kidney tissues were increased; however, its role in diabetic nephropathy remains unclear. In this study, differences in glycation degree and Tf sites induced by differing high glucose concentrations in vitro and the effect on total iron binding capacity (TIBC) were observed. Moreover, the effect of AGE-Tf on human renal tubular epithelial cells (HK-2) was investigated.MethodsIn vitro Tf was incubated with increasing glucose concentrations (0 mM, 5.6 mM, 11.1 mM, 33.3 mM, 100 mM, 500 mM, and 1,000 mM) for AGE-Tf. Differences in AGE-Tf glycation degree and TIBC level were analyzed via colorimetric method. The AGE-Tf glycation sites were identified with LC-MS/MS. HK-2 cells were treated with AGE-Tf prepared with different glucose concentrations (33.3 mM and 500 mM) in vitro. The effects of AGE-Tf on HK-2 cell viability, proliferation, oxidative stress index, and Tf receptor expression levels were then observed.ResultsWith increasing glucose concentrations (100 mM, 500 mM, and 1,000 mM) in vitro, Tf glycation degree was significantly increased. The TIBC levels of AGE-Tf were decreased significantly with increasing glucose concentrations (33.3 mM, 100 mM, 500 mM, and 1,000 mM). Four glycated modification sites in Tf and 17 glycated modification sites were detected in AGE-Tf (500 mM) by LC-MS/MS. The structural types of AGEs were CML, G-H1, FL-1H2O, FL, and MG-H1. No significant differences were found in the survival rate of HK-2 cells among the AGE-Tf (500 mM), AGE-Tf (33.3 mM), and Tf groups (all p > 0.05). The apoptosis rate of HK-2 cells in the AGE-Tf (500 mM) group was significantly higher than that in the AGE-Tf (33.3 mM) group. Additionally, both of them were significantly higher than that in the Tf group (both p < 0.05). The MDA levels of HK-2 cells in the AGE-Tf (500 mM) and AGE-Tf (33.3 mM) groups were higher than that in the Tf group, but not significantly (both p > 0.05). The T-AOC level of HK-2 in the AGE-Tf (500 mM) group was significantly lower than that in the AGE-Tf (33.3 mM) and Tf groups (both p < 0.001). The GSH level of HK-2 cells in the AGE-Tf (500 mM) group was significantly lower than that in the Tf group (p < 0.05). The expression level of TfR in the AGE-Tf (500 mM) group was also significantly lower than that in the Tf group (p < 0.05).ConclusionThe degree and sites of Tf glycation were increased in vitro secondary to high-glucose exposure; however, the binding ability of Tf to iron decreased gradually. After HK-2 was stimulated by AGE-Tf in vitro, the apoptosis of cells was increased, antioxidant capacity was decreased, and TfR expression levels were downregulated. |
| first_indexed | 2024-04-10T20:13:00Z |
| format | Article |
| id | doaj.art-00f766ca51ea4914bb772a2afd46f1ea |
| institution | Directory Open Access Journal |
| issn | 1664-2392 |
| language | English |
| last_indexed | 2024-04-10T20:13:00Z |
| publishDate | 2023-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Endocrinology |
| spelling | doaj.art-00f766ca51ea4914bb772a2afd46f1ea2023-01-26T08:30:49ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922023-01-011310.3389/fendo.2022.10095071009507Effect of transferrin glycation induced by high glucose on HK-2 cells in vitroYanqi Ma0Qikai Zhou1Pingping Zhao2Xiaoyu Lv3Caixia Gong4Jie Gao5Jingfang Liu6Jingfang Liu7The First Clinical Medical College, Lanzhou University, Lanzhou, Gansu, ChinaTianjin Normal University, Tianjin, ChinaThe First Clinical Medical College, Lanzhou University, Lanzhou, Gansu, ChinaThe First Clinical Medical College, Lanzhou University, Lanzhou, Gansu, ChinaThe First Clinical Medical College, Lanzhou University, Lanzhou, Gansu, ChinaThe First Clinical Medical College, Lanzhou University, Lanzhou, Gansu, ChinaThe First Clinical Medical College, Lanzhou University, Lanzhou, Gansu, ChinaDepartment of Endocrinology, The First Hospital of Lanzhou University, Lanzhou, Gansu, ChinaBackground and objectiveGlycation is a common post-transcriptional modification of proteins. Previous studies have shown that advanced glycation end modified transferrin (AGE-Tf) levels in diabetic rat kidney tissues were increased; however, its role in diabetic nephropathy remains unclear. In this study, differences in glycation degree and Tf sites induced by differing high glucose concentrations in vitro and the effect on total iron binding capacity (TIBC) were observed. Moreover, the effect of AGE-Tf on human renal tubular epithelial cells (HK-2) was investigated.MethodsIn vitro Tf was incubated with increasing glucose concentrations (0 mM, 5.6 mM, 11.1 mM, 33.3 mM, 100 mM, 500 mM, and 1,000 mM) for AGE-Tf. Differences in AGE-Tf glycation degree and TIBC level were analyzed via colorimetric method. The AGE-Tf glycation sites were identified with LC-MS/MS. HK-2 cells were treated with AGE-Tf prepared with different glucose concentrations (33.3 mM and 500 mM) in vitro. The effects of AGE-Tf on HK-2 cell viability, proliferation, oxidative stress index, and Tf receptor expression levels were then observed.ResultsWith increasing glucose concentrations (100 mM, 500 mM, and 1,000 mM) in vitro, Tf glycation degree was significantly increased. The TIBC levels of AGE-Tf were decreased significantly with increasing glucose concentrations (33.3 mM, 100 mM, 500 mM, and 1,000 mM). Four glycated modification sites in Tf and 17 glycated modification sites were detected in AGE-Tf (500 mM) by LC-MS/MS. The structural types of AGEs were CML, G-H1, FL-1H2O, FL, and MG-H1. No significant differences were found in the survival rate of HK-2 cells among the AGE-Tf (500 mM), AGE-Tf (33.3 mM), and Tf groups (all p > 0.05). The apoptosis rate of HK-2 cells in the AGE-Tf (500 mM) group was significantly higher than that in the AGE-Tf (33.3 mM) group. Additionally, both of them were significantly higher than that in the Tf group (both p < 0.05). The MDA levels of HK-2 cells in the AGE-Tf (500 mM) and AGE-Tf (33.3 mM) groups were higher than that in the Tf group, but not significantly (both p > 0.05). The T-AOC level of HK-2 in the AGE-Tf (500 mM) group was significantly lower than that in the AGE-Tf (33.3 mM) and Tf groups (both p < 0.001). The GSH level of HK-2 cells in the AGE-Tf (500 mM) group was significantly lower than that in the Tf group (p < 0.05). The expression level of TfR in the AGE-Tf (500 mM) group was also significantly lower than that in the Tf group (p < 0.05).ConclusionThe degree and sites of Tf glycation were increased in vitro secondary to high-glucose exposure; however, the binding ability of Tf to iron decreased gradually. After HK-2 was stimulated by AGE-Tf in vitro, the apoptosis of cells was increased, antioxidant capacity was decreased, and TfR expression levels were downregulated.https://www.frontiersin.org/articles/10.3389/fendo.2022.1009507/fullglycationtransferrinadvanced glycation end modified transferrinHK-2transferrin receptor |
| spellingShingle | Yanqi Ma Qikai Zhou Pingping Zhao Xiaoyu Lv Caixia Gong Jie Gao Jingfang Liu Jingfang Liu Effect of transferrin glycation induced by high glucose on HK-2 cells in vitro Frontiers in Endocrinology glycation transferrin advanced glycation end modified transferrin HK-2 transferrin receptor |
| title | Effect of transferrin glycation induced by high glucose on HK-2 cells in vitro |
| title_full | Effect of transferrin glycation induced by high glucose on HK-2 cells in vitro |
| title_fullStr | Effect of transferrin glycation induced by high glucose on HK-2 cells in vitro |
| title_full_unstemmed | Effect of transferrin glycation induced by high glucose on HK-2 cells in vitro |
| title_short | Effect of transferrin glycation induced by high glucose on HK-2 cells in vitro |
| title_sort | effect of transferrin glycation induced by high glucose on hk 2 cells in vitro |
| topic | glycation transferrin advanced glycation end modified transferrin HK-2 transferrin receptor |
| url | https://www.frontiersin.org/articles/10.3389/fendo.2022.1009507/full |
| work_keys_str_mv | AT yanqima effectoftransferringlycationinducedbyhighglucoseonhk2cellsinvitro AT qikaizhou effectoftransferringlycationinducedbyhighglucoseonhk2cellsinvitro AT pingpingzhao effectoftransferringlycationinducedbyhighglucoseonhk2cellsinvitro AT xiaoyulv effectoftransferringlycationinducedbyhighglucoseonhk2cellsinvitro AT caixiagong effectoftransferringlycationinducedbyhighglucoseonhk2cellsinvitro AT jiegao effectoftransferringlycationinducedbyhighglucoseonhk2cellsinvitro AT jingfangliu effectoftransferringlycationinducedbyhighglucoseonhk2cellsinvitro AT jingfangliu effectoftransferringlycationinducedbyhighglucoseonhk2cellsinvitro |