RIPK2-Mediated Autophagy and Negatively Regulated ROS-NLRP3 Inflammasome Signaling in GMCs Stimulated with High Glucose
Background. Hyperglycemia plays a vital role in diabetic nephropathy (DN); autophagy and its potential upregulator receptor-interacting protein kinase 2 (RIPK2) are associated with ROS, which play a potential role in regulating NLRP3, and may be involved in inflammation in DN. Aim. In this study, we...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2019-01-01
|
| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2019/6207563 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832554291893633024 |
|---|---|
| author | Chenlin Gao Jiao Chen Fang Fan Yang Long Shi Tang Chunxia Jiang Jiying Wang Youhua Xu Yong Xu |
| author_facet | Chenlin Gao Jiao Chen Fang Fan Yang Long Shi Tang Chunxia Jiang Jiying Wang Youhua Xu Yong Xu |
| author_sort | Chenlin Gao |
| collection | DOAJ |
| description | Background. Hyperglycemia plays a vital role in diabetic nephropathy (DN); autophagy and its potential upregulator receptor-interacting protein kinase 2 (RIPK2) are associated with ROS, which play a potential role in regulating NLRP3, and may be involved in inflammation in DN. Aim. In this study, we aimed to explore the mechanisms mediated by RIPK2 in autophagy and the relationship with ROS-NLRP3 of DN, by investigating the levels of RIPK2 and autophagy in glomerular mesangial cells (GMCs) stimulated with high glucose. Material and Methods. GMCs were divided into the following groups: normal group (NC), high glucose group (HG), and RIPK2 siRNA group. RIPK2, LC3, caspase1, and IL-1β levels were measured by western blotting and RT-PCR. Autophagosomes were measured by GFP-RFP-LC3; ROS were detected by DCFH-DA. Results. High glucose upregulated RIPK2 and LC3 in GMCs during short periods (0-12 h) (p<0.01), while RIPK2 and LC3 were significantly downregulated in the long term (12-72 h) (p<0.01); these changes were positively correlated with glucose concentration (p<0.01). In addition, levels of ROS, caspase1, and IL-1β increased in a time- and dose-dependent manner in the high glucose group, even with an increased expression of LC3 (p<0.01). However, LC3 expression decreased in the siRIPK2 group, while levels of ROS, caspase1, and IL-1β increased (p<0.01). Conclusions. Autophagy was activated by high glucose at short time periods but was inhibited in the long term, demonstrating a dual role for high glucose in autophagy of GMCs. RIPK2 regulates ROS-NLRP3 inflammasome signaling through autophagy and may be involved in the pathogenesis of DN. |
| format | Article |
| id | doaj-art-7275e6604227425d9ebe28b370d2d6bf |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-7275e6604227425d9ebe28b370d2d6bf2025-02-03T05:51:55ZengWileyMediators of Inflammation0962-93511466-18612019-01-01201910.1155/2019/62075636207563RIPK2-Mediated Autophagy and Negatively Regulated ROS-NLRP3 Inflammasome Signaling in GMCs Stimulated with High GlucoseChenlin Gao0Jiao Chen1Fang Fan2Yang Long3Shi Tang4Chunxia Jiang5Jiying Wang6Youhua Xu7Yong Xu8Endocrinology Department, The Affiliated Hospital of Southwest Medical University, Sichuan Luzhou 646000, ChinaEndocrinology Department, The Affiliated Hospital of Southwest Medical University, Sichuan Luzhou 646000, ChinaDepartment of Endocrinology, The First People’s Hospital of Neijiang, Sichuan 641000, ChinaEndocrinology Department, The Affiliated Hospital of Southwest Medical University, Sichuan Luzhou 646000, ChinaEndocrinology Department, The Affiliated Hospital of Nuclear Industry 416 Hospital, Chengdu, Sichuan 610000, ChinaEndocrinology Department, The Affiliated Hospital of Southwest Medical University, Sichuan Luzhou 646000, ChinaDepartment of Endocrinology, Guangyuan Central Hospital, Guangyuan, Sichuan 628000, ChinaDepartment of Endocrinology, The First People’s Hospital of Neijiang, Sichuan 641000, ChinaEndocrinology Department, The Affiliated Hospital of Southwest Medical University, Sichuan Luzhou 646000, ChinaBackground. Hyperglycemia plays a vital role in diabetic nephropathy (DN); autophagy and its potential upregulator receptor-interacting protein kinase 2 (RIPK2) are associated with ROS, which play a potential role in regulating NLRP3, and may be involved in inflammation in DN. Aim. In this study, we aimed to explore the mechanisms mediated by RIPK2 in autophagy and the relationship with ROS-NLRP3 of DN, by investigating the levels of RIPK2 and autophagy in glomerular mesangial cells (GMCs) stimulated with high glucose. Material and Methods. GMCs were divided into the following groups: normal group (NC), high glucose group (HG), and RIPK2 siRNA group. RIPK2, LC3, caspase1, and IL-1β levels were measured by western blotting and RT-PCR. Autophagosomes were measured by GFP-RFP-LC3; ROS were detected by DCFH-DA. Results. High glucose upregulated RIPK2 and LC3 in GMCs during short periods (0-12 h) (p<0.01), while RIPK2 and LC3 were significantly downregulated in the long term (12-72 h) (p<0.01); these changes were positively correlated with glucose concentration (p<0.01). In addition, levels of ROS, caspase1, and IL-1β increased in a time- and dose-dependent manner in the high glucose group, even with an increased expression of LC3 (p<0.01). However, LC3 expression decreased in the siRIPK2 group, while levels of ROS, caspase1, and IL-1β increased (p<0.01). Conclusions. Autophagy was activated by high glucose at short time periods but was inhibited in the long term, demonstrating a dual role for high glucose in autophagy of GMCs. RIPK2 regulates ROS-NLRP3 inflammasome signaling through autophagy and may be involved in the pathogenesis of DN.http://dx.doi.org/10.1155/2019/6207563 |
| spellingShingle | Chenlin Gao Jiao Chen Fang Fan Yang Long Shi Tang Chunxia Jiang Jiying Wang Youhua Xu Yong Xu RIPK2-Mediated Autophagy and Negatively Regulated ROS-NLRP3 Inflammasome Signaling in GMCs Stimulated with High Glucose Mediators of Inflammation |
| title | RIPK2-Mediated Autophagy and Negatively Regulated ROS-NLRP3 Inflammasome Signaling in GMCs Stimulated with High Glucose |
| title_full | RIPK2-Mediated Autophagy and Negatively Regulated ROS-NLRP3 Inflammasome Signaling in GMCs Stimulated with High Glucose |
| title_fullStr | RIPK2-Mediated Autophagy and Negatively Regulated ROS-NLRP3 Inflammasome Signaling in GMCs Stimulated with High Glucose |
| title_full_unstemmed | RIPK2-Mediated Autophagy and Negatively Regulated ROS-NLRP3 Inflammasome Signaling in GMCs Stimulated with High Glucose |
| title_short | RIPK2-Mediated Autophagy and Negatively Regulated ROS-NLRP3 Inflammasome Signaling in GMCs Stimulated with High Glucose |
| title_sort | ripk2 mediated autophagy and negatively regulated ros nlrp3 inflammasome signaling in gmcs stimulated with high glucose |
| url | http://dx.doi.org/10.1155/2019/6207563 |
| work_keys_str_mv | AT chenlingao ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose AT jiaochen ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose AT fangfan ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose AT yanglong ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose AT shitang ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose AT chunxiajiang ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose AT jiyingwang ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose AT youhuaxu ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose AT yongxu ripk2mediatedautophagyandnegativelyregulatedrosnlrp3inflammasomesignalingingmcsstimulatedwithhighglucose |