Pristimerin Inhibits MMP-9 Expression and Cell Migration Through Attenuating NOX/ROS-Dependent NF-κB Activation in Rat Brain Astrocytes Challenged with LPS
Chien-Chung Yang,1,2 Li-Der Hsiao,3 Hui-Ching Tseng,3 Ching-Ming Kuo,3 Chuen-Mao Yang3,4 1Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital at Tao-Yuan, Tao-Yuan 33302, Taiwan; 2School of Traditional Chinese Medicine, College of Medicine, Chang Gung University, Tao-Yuan 33302,...
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Dove Medical Press
2020
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oai:doaj.org-article:b7e9109ea9c54e00a653d3155217d5a62021-12-02T10:08:50ZPristimerin Inhibits MMP-9 Expression and Cell Migration Through Attenuating NOX/ROS-Dependent NF-κB Activation in Rat Brain Astrocytes Challenged with LPS1178-7031https://doaj.org/article/b7e9109ea9c54e00a653d3155217d5a62020-07-01T00:00:00Zhttps://www.dovepress.com/pristimerin-inhibits-mmp-9-expression-and-cell-migration-through-atten-peer-reviewed-article-JIRhttps://doaj.org/toc/1178-7031Chien-Chung Yang,1,2 Li-Der Hsiao,3 Hui-Ching Tseng,3 Ching-Ming Kuo,3 Chuen-Mao Yang3,4 1Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital at Tao-Yuan, Tao-Yuan 33302, Taiwan; 2School of Traditional Chinese Medicine, College of Medicine, Chang Gung University, Tao-Yuan 33302, Taiwan; 3Department of Pharmacology, College of Medicine, China Medical University, Taichung 40402, Taiwan; 4Department of Post-Baccalaureate Veterinary Medicine, College of Medical and Health Science, Asia University, Taichung 41354, TaiwanCorrespondence: Chuen-Mao Yang Tel +886 4-22053366 (Ext. 2229)Email chuenmao@mail.cmu.edu.twPurpose: Neuroinflammation plays a crucial role in neurodegenerative diseases. Matrix metalloproteinases (MMPs) are a landmark of neuroinflammation. Lipopolysaccharide (LPS) has been demonstrated to induce MMP-9 expression. The mechanisms underlying LPS-induced MMP-9 expression have not been completely elucidated in astrocytes. Nuclear factor-kappaB (NF-κB) is well known as one of the crucial transcription factors in MMP-9 induction. Moreover, reactive oxygen species (ROS) could be an important mediator of neuroinflammation. Here, we differentiated whether ROS and NF-κB contributed to LPS-mediated MMP-9 expression in rat brain astrocytes (RBA-1). Besides, pristimerin has been revealed to possess antioxidant and anti-inflammatory effects. We also evaluated the effects of pristimerin on LPS-induced inflammatory responses.Methods: RBA-1 cells were used for analyses. Pharmacological inhibitors and siRNAs were used to evaluate the signaling pathway. Western blotting and gelatin zymography were conducted to evaluate protein and MMP-9 expression, respectively. Real-time PCR was for mRNA expression. Wound healing assay was for cell migration. 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate (H2DCF-DA) and dihydroethidium (DHE) staining were for ROS generation. Immunofluorescence staining was conducted to assess NF-κB p65. Promoter-reporter gene assay and chromatin immunoprecipitation (ChIP) assay were used to detect promoter activity and the association of nuclear proteins with the promoter.Results: Our results showed that the increased level of ROS generation was attenuated by edaravone (a ROS scavenger), apocynin (APO; an inhibitor of p47Phox), diphenyleneiodonium (DPI; an inhibitor of NOX), and pristimerin in RBA-1 cells exposed to LPS. Besides, pretreatment with APO, DPI, edaravone, Bay11-7082, and pristimerin also inhibited the phosphorylation, nuclear translocation, promoter binding activity of NF-κB p65 as well as upregulation of MMP-9 expression-mediated cell migration in RBA-1 cells challenged with LPS.Conclusion: These results suggested that LPS enhances the upregulation of MMP-9 through nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX)/ROS-dependent NF-κB activity. These results also provide new insights into the mechanisms by which pristimerin attenuates LPS-mediated MMP-9 expression and neuroinflammatory responses.Keywords: neuroinflammation, NOX, ROS, LPS, MMP-9, pristimerin, astrocytesYang CCHsiao LDTseng HCKuo CMYang CMDove Medical Pressarticleneuroinflammationnoxroslpsmmp-9pristimerinastrocytesPathologyRB1-214Therapeutics. PharmacologyRM1-950ENJournal of Inflammation Research, Vol Volume 13, Pp 325-341 (2020) |
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DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
neuroinflammation nox ros lps mmp-9 pristimerin astrocytes Pathology RB1-214 Therapeutics. Pharmacology RM1-950 |
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neuroinflammation nox ros lps mmp-9 pristimerin astrocytes Pathology RB1-214 Therapeutics. Pharmacology RM1-950 Yang CC Hsiao LD Tseng HC Kuo CM Yang CM Pristimerin Inhibits MMP-9 Expression and Cell Migration Through Attenuating NOX/ROS-Dependent NF-κB Activation in Rat Brain Astrocytes Challenged with LPS |
| description |
Chien-Chung Yang,1,2 Li-Der Hsiao,3 Hui-Ching Tseng,3 Ching-Ming Kuo,3 Chuen-Mao Yang3,4 1Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital at Tao-Yuan, Tao-Yuan 33302, Taiwan; 2School of Traditional Chinese Medicine, College of Medicine, Chang Gung University, Tao-Yuan 33302, Taiwan; 3Department of Pharmacology, College of Medicine, China Medical University, Taichung 40402, Taiwan; 4Department of Post-Baccalaureate Veterinary Medicine, College of Medical and Health Science, Asia University, Taichung 41354, TaiwanCorrespondence: Chuen-Mao Yang Tel +886 4-22053366 (Ext. 2229)Email chuenmao@mail.cmu.edu.twPurpose: Neuroinflammation plays a crucial role in neurodegenerative diseases. Matrix metalloproteinases (MMPs) are a landmark of neuroinflammation. Lipopolysaccharide (LPS) has been demonstrated to induce MMP-9 expression. The mechanisms underlying LPS-induced MMP-9 expression have not been completely elucidated in astrocytes. Nuclear factor-kappaB (NF-κB) is well known as one of the crucial transcription factors in MMP-9 induction. Moreover, reactive oxygen species (ROS) could be an important mediator of neuroinflammation. Here, we differentiated whether ROS and NF-κB contributed to LPS-mediated MMP-9 expression in rat brain astrocytes (RBA-1). Besides, pristimerin has been revealed to possess antioxidant and anti-inflammatory effects. We also evaluated the effects of pristimerin on LPS-induced inflammatory responses.Methods: RBA-1 cells were used for analyses. Pharmacological inhibitors and siRNAs were used to evaluate the signaling pathway. Western blotting and gelatin zymography were conducted to evaluate protein and MMP-9 expression, respectively. Real-time PCR was for mRNA expression. Wound healing assay was for cell migration. 2ʹ,7ʹ-dichlorodihydrofluorescein diacetate (H2DCF-DA) and dihydroethidium (DHE) staining were for ROS generation. Immunofluorescence staining was conducted to assess NF-κB p65. Promoter-reporter gene assay and chromatin immunoprecipitation (ChIP) assay were used to detect promoter activity and the association of nuclear proteins with the promoter.Results: Our results showed that the increased level of ROS generation was attenuated by edaravone (a ROS scavenger), apocynin (APO; an inhibitor of p47Phox), diphenyleneiodonium (DPI; an inhibitor of NOX), and pristimerin in RBA-1 cells exposed to LPS. Besides, pretreatment with APO, DPI, edaravone, Bay11-7082, and pristimerin also inhibited the phosphorylation, nuclear translocation, promoter binding activity of NF-κB p65 as well as upregulation of MMP-9 expression-mediated cell migration in RBA-1 cells challenged with LPS.Conclusion: These results suggested that LPS enhances the upregulation of MMP-9 through nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX)/ROS-dependent NF-κB activity. These results also provide new insights into the mechanisms by which pristimerin attenuates LPS-mediated MMP-9 expression and neuroinflammatory responses.Keywords: neuroinflammation, NOX, ROS, LPS, MMP-9, pristimerin, astrocytes |
| format |
article |
| author |
Yang CC Hsiao LD Tseng HC Kuo CM Yang CM |
| author_facet |
Yang CC Hsiao LD Tseng HC Kuo CM Yang CM |
| author_sort |
Yang CC |
| title |
Pristimerin Inhibits MMP-9 Expression and Cell Migration Through Attenuating NOX/ROS-Dependent NF-κB Activation in Rat Brain Astrocytes Challenged with LPS |
| title_short |
Pristimerin Inhibits MMP-9 Expression and Cell Migration Through Attenuating NOX/ROS-Dependent NF-κB Activation in Rat Brain Astrocytes Challenged with LPS |
| title_full |
Pristimerin Inhibits MMP-9 Expression and Cell Migration Through Attenuating NOX/ROS-Dependent NF-κB Activation in Rat Brain Astrocytes Challenged with LPS |
| title_fullStr |
Pristimerin Inhibits MMP-9 Expression and Cell Migration Through Attenuating NOX/ROS-Dependent NF-κB Activation in Rat Brain Astrocytes Challenged with LPS |
| title_full_unstemmed |
Pristimerin Inhibits MMP-9 Expression and Cell Migration Through Attenuating NOX/ROS-Dependent NF-κB Activation in Rat Brain Astrocytes Challenged with LPS |
| title_sort |
pristimerin inhibits mmp-9 expression and cell migration through attenuating nox/ros-dependent nf-κb activation in rat brain astrocytes challenged with lps |
| publisher |
Dove Medical Press |
| publishDate |
2020 |
| url |
https://doaj.org/article/b7e9109ea9c54e00a653d3155217d5a6 |
| work_keys_str_mv |
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