A Novel Phytochemical, DIM, Inhibits Proliferation, Migration, Invasion and TNF-α Induced Inflammatory Cytokine Production of Synovial Fibroblasts From Rheumatoid Arthritis Patients by Targeting MAPK and AKT/mTOR Signal Pathway
In rheumatoid arthritis(RA) pathogenesis, activated RA fibroblast-like synoviocytes (RA-FLSs) exhibit similar proliferative features as tumor cells and subsequent erosion to cartilage will eventually lead to joint destruction. Therefore, it is imperative to search for compounds, which can effectivel...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2019-07-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2019.01620/full |
| _version_ | 1818960558752268288 |
|---|---|
| author | Hongyan Du Xi Zhang Yongchang Zeng Xiaoming Huang Hao Chen Suihai Wang Jing Wu Qiang Li Wei Zhu Hongwei Li Tiancai Liu Qinghong Yu Yingsong Wu Ligang Jie |
| author_facet | Hongyan Du Xi Zhang Yongchang Zeng Xiaoming Huang Hao Chen Suihai Wang Jing Wu Qiang Li Wei Zhu Hongwei Li Tiancai Liu Qinghong Yu Yingsong Wu Ligang Jie |
| author_sort | Hongyan Du |
| collection | DOAJ |
| description | In rheumatoid arthritis(RA) pathogenesis, activated RA fibroblast-like synoviocytes (RA-FLSs) exhibit similar proliferative features as tumor cells and subsequent erosion to cartilage will eventually lead to joint destruction. Therefore, it is imperative to search for compounds, which can effectively inhibit the abnormal activation of RA-FLSs, and retard RA progression.3′3-Diindolylmethane (DIM), the major product of the acid-catalyzed oligomerization of indole-3-carbinol from cruciferous vegetables, has been reported to be functionally relevant to inhibition of migration, invasion and carcinogenesis in some solid tumors. In this study, we explored the anti-proliferation, anti-metastasis and anti-inflammation effects of DIM on RA-FLSs as well as the underlying molecular mechanisms. To do this, primary RA-FLSs were isolated from RA patients and an animal model. Cell proliferation, migration and invasion were measured using CCK-8, scratch, and Transwell assays, respectively. The effects of DIM on Matrix metalloproteinases (MMPs) and some inflammatory factors mRNA and key molecules such as some inflammatory factors and those involved in aberrantly-activated signaling pathway in response to tumor necrosis factor α(TNF-α), a typical characteristic mediator in RA-FLS, were quantitatively measured by real-time PCR and western blotting. Moreover, the effect of DIM on adjuvant induced arthritis(AIA) models was evaluated with C57BL/6 mice in vivo. The results showed that DIM inhibited proliferation, migration and invasion of RA-FLS in vitro. Meanwhile, DIM dramatically suppressed TNF-α–induced increases in the mRNA levels of MMP-2, MMP-3, MMP-8, and MMP-9; as well as the proinflammatory factors IL-6, IL-8, and IL-1β. Mechanistic studies revealed that DIM is able to suppress phosphorylated activation not only of p38, JNK in MAPK pathway but of AKT, mTOR and downstream molecules in the AKT/mTOR pathway. Moreover, DIM treatment decreased expression levels of proinflammatory cytokines in the serum and alleviated arthritis severity in the knee joints of AIA mice. Taken together, our findings demonstrate that DIM could inhibit proliferation, migration and invasion of RA-FLSs and reduce proinflammatory factors induced by TNF-α in vitro by blocking MAPK and AKT/mTOR pathway and prevent inflammation and knee joint destruction in vivo, which suggests that DIM might have therapeutic potential for RA. |
| first_indexed | 2024-12-20T11:59:27Z |
| format | Article |
| id | doaj.art-90dcecf8dd1245f299162579e1f57574 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-20T11:59:27Z |
| publishDate | 2019-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-90dcecf8dd1245f299162579e1f575742022-12-21T19:41:36ZengFrontiers Media S.A.Frontiers in Immunology1664-32242019-07-011010.3389/fimmu.2019.01620459630A Novel Phytochemical, DIM, Inhibits Proliferation, Migration, Invasion and TNF-α Induced Inflammatory Cytokine Production of Synovial Fibroblasts From Rheumatoid Arthritis Patients by Targeting MAPK and AKT/mTOR Signal PathwayHongyan Du0Xi Zhang1Yongchang Zeng2Xiaoming Huang3Hao Chen4Suihai Wang5Jing Wu6Qiang Li7Wei Zhu8Hongwei Li9Tiancai Liu10Qinghong Yu11Yingsong Wu12Ligang Jie13School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaSchool of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaSchool of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaSchool of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaSchool of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaSchool of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaDepartment of Rheumatology and Clinical Immunology, Zhujiang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Rheumatology and Clinical Immunology, Zhujiang Hospital, Southern Medical University, Guangzhou, ChinaDepartment of Toxicology, Guangzhou Center for Disease Control and Prevention, Guangzhou, ChinaSchool of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaSchool of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaDepartment of Rheumatology and Clinical Immunology, Zhujiang Hospital, Southern Medical University, Guangzhou, ChinaSchool of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, ChinaDepartment of Rheumatology and Clinical Immunology, Zhujiang Hospital, Southern Medical University, Guangzhou, ChinaIn rheumatoid arthritis(RA) pathogenesis, activated RA fibroblast-like synoviocytes (RA-FLSs) exhibit similar proliferative features as tumor cells and subsequent erosion to cartilage will eventually lead to joint destruction. Therefore, it is imperative to search for compounds, which can effectively inhibit the abnormal activation of RA-FLSs, and retard RA progression.3′3-Diindolylmethane (DIM), the major product of the acid-catalyzed oligomerization of indole-3-carbinol from cruciferous vegetables, has been reported to be functionally relevant to inhibition of migration, invasion and carcinogenesis in some solid tumors. In this study, we explored the anti-proliferation, anti-metastasis and anti-inflammation effects of DIM on RA-FLSs as well as the underlying molecular mechanisms. To do this, primary RA-FLSs were isolated from RA patients and an animal model. Cell proliferation, migration and invasion were measured using CCK-8, scratch, and Transwell assays, respectively. The effects of DIM on Matrix metalloproteinases (MMPs) and some inflammatory factors mRNA and key molecules such as some inflammatory factors and those involved in aberrantly-activated signaling pathway in response to tumor necrosis factor α(TNF-α), a typical characteristic mediator in RA-FLS, were quantitatively measured by real-time PCR and western blotting. Moreover, the effect of DIM on adjuvant induced arthritis(AIA) models was evaluated with C57BL/6 mice in vivo. The results showed that DIM inhibited proliferation, migration and invasion of RA-FLS in vitro. Meanwhile, DIM dramatically suppressed TNF-α–induced increases in the mRNA levels of MMP-2, MMP-3, MMP-8, and MMP-9; as well as the proinflammatory factors IL-6, IL-8, and IL-1β. Mechanistic studies revealed that DIM is able to suppress phosphorylated activation not only of p38, JNK in MAPK pathway but of AKT, mTOR and downstream molecules in the AKT/mTOR pathway. Moreover, DIM treatment decreased expression levels of proinflammatory cytokines in the serum and alleviated arthritis severity in the knee joints of AIA mice. Taken together, our findings demonstrate that DIM could inhibit proliferation, migration and invasion of RA-FLSs and reduce proinflammatory factors induced by TNF-α in vitro by blocking MAPK and AKT/mTOR pathway and prevent inflammation and knee joint destruction in vivo, which suggests that DIM might have therapeutic potential for RA.https://www.frontiersin.org/article/10.3389/fimmu.2019.01620/full3′3-Diindolylmethane(DIM)suppressrheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs)adjuvant-induced arthritis (AIA)MAPKAKT/mTOR |
| spellingShingle | Hongyan Du Xi Zhang Yongchang Zeng Xiaoming Huang Hao Chen Suihai Wang Jing Wu Qiang Li Wei Zhu Hongwei Li Tiancai Liu Qinghong Yu Yingsong Wu Ligang Jie A Novel Phytochemical, DIM, Inhibits Proliferation, Migration, Invasion and TNF-α Induced Inflammatory Cytokine Production of Synovial Fibroblasts From Rheumatoid Arthritis Patients by Targeting MAPK and AKT/mTOR Signal Pathway Frontiers in Immunology 3′3-Diindolylmethane(DIM) suppress rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) adjuvant-induced arthritis (AIA) MAPK AKT/mTOR |
| title | A Novel Phytochemical, DIM, Inhibits Proliferation, Migration, Invasion and TNF-α Induced Inflammatory Cytokine Production of Synovial Fibroblasts From Rheumatoid Arthritis Patients by Targeting MAPK and AKT/mTOR Signal Pathway |
| title_full | A Novel Phytochemical, DIM, Inhibits Proliferation, Migration, Invasion and TNF-α Induced Inflammatory Cytokine Production of Synovial Fibroblasts From Rheumatoid Arthritis Patients by Targeting MAPK and AKT/mTOR Signal Pathway |
| title_fullStr | A Novel Phytochemical, DIM, Inhibits Proliferation, Migration, Invasion and TNF-α Induced Inflammatory Cytokine Production of Synovial Fibroblasts From Rheumatoid Arthritis Patients by Targeting MAPK and AKT/mTOR Signal Pathway |
| title_full_unstemmed | A Novel Phytochemical, DIM, Inhibits Proliferation, Migration, Invasion and TNF-α Induced Inflammatory Cytokine Production of Synovial Fibroblasts From Rheumatoid Arthritis Patients by Targeting MAPK and AKT/mTOR Signal Pathway |
| title_short | A Novel Phytochemical, DIM, Inhibits Proliferation, Migration, Invasion and TNF-α Induced Inflammatory Cytokine Production of Synovial Fibroblasts From Rheumatoid Arthritis Patients by Targeting MAPK and AKT/mTOR Signal Pathway |
| title_sort | novel phytochemical dim inhibits proliferation migration invasion and tnf α induced inflammatory cytokine production of synovial fibroblasts from rheumatoid arthritis patients by targeting mapk and akt mtor signal pathway |
| topic | 3′3-Diindolylmethane(DIM) suppress rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) adjuvant-induced arthritis (AIA) MAPK AKT/mTOR |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2019.01620/full |
| work_keys_str_mv | AT hongyandu anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT xizhang anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT yongchangzeng anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT xiaominghuang anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT haochen anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT suihaiwang anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT jingwu anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT qiangli anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT weizhu anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT hongweili anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT tiancailiu anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT qinghongyu anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT yingsongwu anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT ligangjie anovelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT hongyandu novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT xizhang novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT yongchangzeng novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT xiaominghuang novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT haochen novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT suihaiwang novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT jingwu novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT qiangli novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT weizhu novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT hongweili novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT tiancailiu novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT qinghongyu novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT yingsongwu novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway AT ligangjie novelphytochemicaldiminhibitsproliferationmigrationinvasionandtnfainducedinflammatorycytokineproductionofsynovialfibroblastsfromrheumatoidarthritispatientsbytargetingmapkandaktmtorsignalpathway |