Lentivirus-Mediated Missense Mutation in HtrA1 Leads to Activation of the TGF-β/Smads Pathway and Increased Apoptosis of Mouse Brain Microvascular Endothelial Cells via the Oxidative Stress Pathway
Background: The aim of this study was to investigate the possible molecular mechanisms underlying cerebral small vessel disease caused by a missense mutation in the high-temperature serine peptidase A1 gene, HtrA1 (NM_002775.4, Exon4, c.905G>A, p.Arg302Gln). Stable strain model...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
IMR Press
2024-10-01
|
| Series: | Journal of Integrative Neuroscience |
| Subjects: | |
| Online Access: | https://www.imrpress.com/journal/JIN/23/11/10.31083/j.jin2311201 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850219047312949248 |
|---|---|
| author | Shi-na Song Hui-juan Li Jian-lin Liang Qian-qian Ren Chang-xin Li Sui-yi Xu |
| author_facet | Shi-na Song Hui-juan Li Jian-lin Liang Qian-qian Ren Chang-xin Li Sui-yi Xu |
| author_sort | Shi-na Song |
| collection | DOAJ |
| description | Background: The aim of this study was to investigate the possible molecular mechanisms underlying cerebral small vessel disease caused by a missense mutation in the high-temperature serine peptidase A1 gene, HtrA1 (NM_002775.4, Exon4, c.905G>A, p.Arg302Gln). Stable strain models were constructed using wild-type and mutant HtrA1 overexpression lentiviral vectors to infect mouse brain microvascular endothelial cells (bEnd.3 cells). Methods: HtrA1 mRNA and protein expression were analyzed by Western blot and quantitative real-time polymerase chain reaction. Western blot technique was also used to evaluate the expression of transforming growth factor (TGF)-β/Smads-related signaling pathway proteins and the oxidative stress pathway protein nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4). The level of reactive oxygen species (ROS) was evaluated using dichloro-dihydro-fluorescein diacetate (DCFH-DA) fluorescent probes. Results: HtrA1 mRNA and protein expression levels were found to be decreased in mutant cells, whereas the levels of ROS, the TGF-β/Smads proteins, and the caspase3 and cleaved-caspase3 apoptotic proteins were increased. Conclusions: Lentivirus-mediated missense mutation in HtrA1 leads to activation of the TGF-β/Smads pathway and to increased apoptosis of mouse brain microvascular endothelial cells via the oxidative stress pathway. Further in vivo studies are required to explore the connections between different signaling pathways in animals, and to identify potential molecular targets for clinical therapy. |
| format | Article |
| id | doaj-art-7c47db2a3fc44ffdbf52f27c66bf223d |
| institution | OA Journals |
| issn | 0219-6352 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | IMR Press |
| record_format | Article |
| series | Journal of Integrative Neuroscience |
| spelling | doaj-art-7c47db2a3fc44ffdbf52f27c66bf223d2025-08-20T02:07:31ZengIMR PressJournal of Integrative Neuroscience0219-63522024-10-01231120110.31083/j.jin2311201S0219-6352(24)00813-1Lentivirus-Mediated Missense Mutation in HtrA1 Leads to Activation of the TGF-β/Smads Pathway and Increased Apoptosis of Mouse Brain Microvascular Endothelial Cells via the Oxidative Stress PathwayShi-na Song0Hui-juan Li1Jian-lin Liang2Qian-qian Ren3Chang-xin Li4Sui-yi Xu5Department of Neurology, Headache Center, The First Hospital of Shanxi Medical University, 030001 Taiyuan, Shanxi, ChinaDepartment of Neurology, Headache Center, The First Hospital of Shanxi Medical University, 030001 Taiyuan, Shanxi, ChinaDepartment of Neurology, Headache Center, The First Hospital of Shanxi Medical University, 030001 Taiyuan, Shanxi, ChinaDepartment of Neurology, Headache Center, The First Hospital of Shanxi Medical University, 030001 Taiyuan, Shanxi, ChinaDepartment of Neurology, Headache Center, The First Hospital of Shanxi Medical University, 030001 Taiyuan, Shanxi, ChinaDepartment of Neurology, Headache Center, The First Hospital of Shanxi Medical University, 030001 Taiyuan, Shanxi, ChinaBackground: The aim of this study was to investigate the possible molecular mechanisms underlying cerebral small vessel disease caused by a missense mutation in the high-temperature serine peptidase A1 gene, HtrA1 (NM_002775.4, Exon4, c.905G>A, p.Arg302Gln). Stable strain models were constructed using wild-type and mutant HtrA1 overexpression lentiviral vectors to infect mouse brain microvascular endothelial cells (bEnd.3 cells). Methods: HtrA1 mRNA and protein expression were analyzed by Western blot and quantitative real-time polymerase chain reaction. Western blot technique was also used to evaluate the expression of transforming growth factor (TGF)-β/Smads-related signaling pathway proteins and the oxidative stress pathway protein nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4). The level of reactive oxygen species (ROS) was evaluated using dichloro-dihydro-fluorescein diacetate (DCFH-DA) fluorescent probes. Results: HtrA1 mRNA and protein expression levels were found to be decreased in mutant cells, whereas the levels of ROS, the TGF-β/Smads proteins, and the caspase3 and cleaved-caspase3 apoptotic proteins were increased. Conclusions: Lentivirus-mediated missense mutation in HtrA1 leads to activation of the TGF-β/Smads pathway and to increased apoptosis of mouse brain microvascular endothelial cells via the oxidative stress pathway. Further in vivo studies are required to explore the connections between different signaling pathways in animals, and to identify potential molecular targets for clinical therapy.https://www.imrpress.com/journal/JIN/23/11/10.31083/j.jin2311201htra1bend.3tgf-β/smadsoxidative stress |
| spellingShingle | Shi-na Song Hui-juan Li Jian-lin Liang Qian-qian Ren Chang-xin Li Sui-yi Xu Lentivirus-Mediated Missense Mutation in HtrA1 Leads to Activation of the TGF-β/Smads Pathway and Increased Apoptosis of Mouse Brain Microvascular Endothelial Cells via the Oxidative Stress Pathway Journal of Integrative Neuroscience htra1 bend.3 tgf-β/smads oxidative stress |
| title | Lentivirus-Mediated Missense Mutation in HtrA1 Leads to Activation of the TGF-β/Smads Pathway and Increased Apoptosis of Mouse Brain Microvascular Endothelial Cells via the Oxidative Stress Pathway |
| title_full | Lentivirus-Mediated Missense Mutation in HtrA1 Leads to Activation of the TGF-β/Smads Pathway and Increased Apoptosis of Mouse Brain Microvascular Endothelial Cells via the Oxidative Stress Pathway |
| title_fullStr | Lentivirus-Mediated Missense Mutation in HtrA1 Leads to Activation of the TGF-β/Smads Pathway and Increased Apoptosis of Mouse Brain Microvascular Endothelial Cells via the Oxidative Stress Pathway |
| title_full_unstemmed | Lentivirus-Mediated Missense Mutation in HtrA1 Leads to Activation of the TGF-β/Smads Pathway and Increased Apoptosis of Mouse Brain Microvascular Endothelial Cells via the Oxidative Stress Pathway |
| title_short | Lentivirus-Mediated Missense Mutation in HtrA1 Leads to Activation of the TGF-β/Smads Pathway and Increased Apoptosis of Mouse Brain Microvascular Endothelial Cells via the Oxidative Stress Pathway |
| title_sort | lentivirus mediated missense mutation in htra1 leads to activation of the tgf β smads pathway and increased apoptosis of mouse brain microvascular endothelial cells via the oxidative stress pathway |
| topic | htra1 bend.3 tgf-β/smads oxidative stress |
| url | https://www.imrpress.com/journal/JIN/23/11/10.31083/j.jin2311201 |
| work_keys_str_mv | AT shinasong lentivirusmediatedmissensemutationinhtra1leadstoactivationofthetgfbsmadspathwayandincreasedapoptosisofmousebrainmicrovascularendothelialcellsviatheoxidativestresspathway AT huijuanli lentivirusmediatedmissensemutationinhtra1leadstoactivationofthetgfbsmadspathwayandincreasedapoptosisofmousebrainmicrovascularendothelialcellsviatheoxidativestresspathway AT jianlinliang lentivirusmediatedmissensemutationinhtra1leadstoactivationofthetgfbsmadspathwayandincreasedapoptosisofmousebrainmicrovascularendothelialcellsviatheoxidativestresspathway AT qianqianren lentivirusmediatedmissensemutationinhtra1leadstoactivationofthetgfbsmadspathwayandincreasedapoptosisofmousebrainmicrovascularendothelialcellsviatheoxidativestresspathway AT changxinli lentivirusmediatedmissensemutationinhtra1leadstoactivationofthetgfbsmadspathwayandincreasedapoptosisofmousebrainmicrovascularendothelialcellsviatheoxidativestresspathway AT suiyixu lentivirusmediatedmissensemutationinhtra1leadstoactivationofthetgfbsmadspathwayandincreasedapoptosisofmousebrainmicrovascularendothelialcellsviatheoxidativestresspathway |