ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent Exosomes
Background. Apoptosis signal-regulating kinase 1 (ASK1) has been reported to induce fibrotic signaling in the setting of oxidative stress. However, the role of ASK1 and its mechanism of action in angiotensin II- (Ang II-) induced liver fibrosis remain largely unknown. Methods. Human hepatic LX-2 ste...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2020-01-01
|
| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2020/8183713 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849691555217014784 |
|---|---|
| author | Pei-pei Fang Chen-wei Pan Wei Lin Jie Li Shan-shan Huang Guang-yao Zhou Wen-jun Du Qiang Li |
| author_facet | Pei-pei Fang Chen-wei Pan Wei Lin Jie Li Shan-shan Huang Guang-yao Zhou Wen-jun Du Qiang Li |
| author_sort | Pei-pei Fang |
| collection | DOAJ |
| description | Background. Apoptosis signal-regulating kinase 1 (ASK1) has been reported to induce fibrotic signaling in the setting of oxidative stress. However, the role of ASK1 and its mechanism of action in angiotensin II- (Ang II-) induced liver fibrosis remain largely unknown. Methods. Human hepatic LX-2 stellate cells were treated with Ang II alone or cotreated with Ang II plus an ASK1 inhibitor (GS-4997) or siRNA-targeting ASK1. Immunofluorescent staining, real-time PCR, and western blotting were used to determine the expressionof α-SMA, Col I, and Col III expression. Cell viability was assessed by the CCK-8 assay. The concentrations of IL-1β, IL-18, and TNF-α in conditioned medium were determined by ELISA. The levels of intracellular ROS in LX-2 cells were analyzed using a ROS assay kit. Exosome size was determined by electron microscopy. Results. Ang II markedly increased the expression of extracellular matrix (ECM) proteins (α-SMA, Col I, and Col III) and proinflammatory cytokines (IL-1β, IL-18, and TNF-α). Ang II also increased the expression of endoplasmic reticulum stress (ERS) markers (GRP78, p-PERK, and CHOP) and p-ASK1. Results also showed that pretreatment with GS-4997 or siRNA could abolish all the abovementioned effects on LX-2 cells. Furthermore, we found that exosome release caused by ASK1-mediated ERS was involved in the activation of LX-2 cells by Ang II. The activation of LX-2 cells could be blocked by treating the exosomes with annexin. Conclusions. In summary, we found that ASK1 mediates Ang II-activated ERS in HSCs and the subsequent activation of HSCs, suggesting a promising strategy for treating liver fibrosis. |
| format | Article |
| id | doaj-art-e1cee5c3cde040deac034ed2e3424294 |
| institution | DOAJ |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-e1cee5c3cde040deac034ed2e34242942025-08-20T03:20:59ZengWileyMediators of Inflammation0962-93511466-18612020-01-01202010.1155/2020/81837138183713ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent ExosomesPei-pei Fang0Chen-wei Pan1Wei Lin2Jie Li3Shan-shan Huang4Guang-yao Zhou5Wen-jun Du6Qiang Li7Division of Liver Diseases, Jinan Infectious Disease Hospital, Shandong University, ChinaDepartment of Infectious Disease, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 325000, ChinaDepartment of Infectious Disease, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 325000, ChinaDepartment of Infectious Disease, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 325000, ChinaDepartment of Infectious Disease, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 325000, ChinaDepartment of Infectious Disease, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 325000, ChinaDivision of Liver Diseases, Jinan Infectious Disease Hospital, Shandong University, ChinaDivision of Liver Diseases, Jinan Infectious Disease Hospital, Shandong University, ChinaBackground. Apoptosis signal-regulating kinase 1 (ASK1) has been reported to induce fibrotic signaling in the setting of oxidative stress. However, the role of ASK1 and its mechanism of action in angiotensin II- (Ang II-) induced liver fibrosis remain largely unknown. Methods. Human hepatic LX-2 stellate cells were treated with Ang II alone or cotreated with Ang II plus an ASK1 inhibitor (GS-4997) or siRNA-targeting ASK1. Immunofluorescent staining, real-time PCR, and western blotting were used to determine the expressionof α-SMA, Col I, and Col III expression. Cell viability was assessed by the CCK-8 assay. The concentrations of IL-1β, IL-18, and TNF-α in conditioned medium were determined by ELISA. The levels of intracellular ROS in LX-2 cells were analyzed using a ROS assay kit. Exosome size was determined by electron microscopy. Results. Ang II markedly increased the expression of extracellular matrix (ECM) proteins (α-SMA, Col I, and Col III) and proinflammatory cytokines (IL-1β, IL-18, and TNF-α). Ang II also increased the expression of endoplasmic reticulum stress (ERS) markers (GRP78, p-PERK, and CHOP) and p-ASK1. Results also showed that pretreatment with GS-4997 or siRNA could abolish all the abovementioned effects on LX-2 cells. Furthermore, we found that exosome release caused by ASK1-mediated ERS was involved in the activation of LX-2 cells by Ang II. The activation of LX-2 cells could be blocked by treating the exosomes with annexin. Conclusions. In summary, we found that ASK1 mediates Ang II-activated ERS in HSCs and the subsequent activation of HSCs, suggesting a promising strategy for treating liver fibrosis.http://dx.doi.org/10.1155/2020/8183713 |
| spellingShingle | Pei-pei Fang Chen-wei Pan Wei Lin Jie Li Shan-shan Huang Guang-yao Zhou Wen-jun Du Qiang Li ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent Exosomes Mediators of Inflammation |
| title | ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent Exosomes |
| title_full | ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent Exosomes |
| title_fullStr | ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent Exosomes |
| title_full_unstemmed | ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent Exosomes |
| title_short | ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent Exosomes |
| title_sort | ask1 enhances angiotensin ii induced liver fibrosis in vitro by mediating endoplasmic reticulum stress dependent exosomes |
| url | http://dx.doi.org/10.1155/2020/8183713 |
| work_keys_str_mv | AT peipeifang ask1enhancesangiotensiniiinducedliverfibrosisinvitrobymediatingendoplasmicreticulumstressdependentexosomes AT chenweipan ask1enhancesangiotensiniiinducedliverfibrosisinvitrobymediatingendoplasmicreticulumstressdependentexosomes AT weilin ask1enhancesangiotensiniiinducedliverfibrosisinvitrobymediatingendoplasmicreticulumstressdependentexosomes AT jieli ask1enhancesangiotensiniiinducedliverfibrosisinvitrobymediatingendoplasmicreticulumstressdependentexosomes AT shanshanhuang ask1enhancesangiotensiniiinducedliverfibrosisinvitrobymediatingendoplasmicreticulumstressdependentexosomes AT guangyaozhou ask1enhancesangiotensiniiinducedliverfibrosisinvitrobymediatingendoplasmicreticulumstressdependentexosomes AT wenjundu ask1enhancesangiotensiniiinducedliverfibrosisinvitrobymediatingendoplasmicreticulumstressdependentexosomes AT qiangli ask1enhancesangiotensiniiinducedliverfibrosisinvitrobymediatingendoplasmicreticulumstressdependentexosomes |