TRAP-induced PAR1 expression with its mechanism during AMI in a rat model
Abstract Background and objective Protease-activated receptor 1 (PAR1) is crucial in individuals with acute myocardial infarction (AMI). The continuous and prompt PAR1 activation mainly dependent on PAR1 trafficking is essential for the role of PAR1 during AMI in which cardiomyocytes are in hypoxia....
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-02-01
|
| Series: | BMC Cardiovascular Disorders |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12872-023-03118-w |
| _version_ | 1797865640123432960 |
|---|---|
| author | Ani Wang Xinyuan Gu Chunyang Wang Yanhui Li Fuhong Deng Jie Fang Naxia Chen Qifu Li Lilong Tang |
| author_facet | Ani Wang Xinyuan Gu Chunyang Wang Yanhui Li Fuhong Deng Jie Fang Naxia Chen Qifu Li Lilong Tang |
| author_sort | Ani Wang |
| collection | DOAJ |
| description | Abstract Background and objective Protease-activated receptor 1 (PAR1) is crucial in individuals with acute myocardial infarction (AMI). The continuous and prompt PAR1 activation mainly dependent on PAR1 trafficking is essential for the role of PAR1 during AMI in which cardiomyocytes are in hypoxia. However, the PAR1 trafficking in cardiomyocytes specially during the hypoxia is still unclear. Methods and result A rat AMI model was created. PAR1 activation with thrombin-receptor activated peptide (TRAP) had a transient effect on cardiac function in normal rats but persistent improvement in rats with AMI. Cardiomyocytes from neonatal rats were cultured in a normal CO2 incubator and a hypoxic modular incubator chamber. The cells were then subjected to western blot for the total protein expression and staining with fluorescent reagent and antibody for PAR1 localization. No change in total PAR1 expression following TRAP stimulation was observed; however, it led to increased PAR1 expression in the early endosomes in normoxic cells and decreased expression in the early endosomes in hypoxic cells. Under hypoxic conditions, TRAP restored the PAR1 expression on both cell and endosomal surfaces within an hour by decreasing Rab11A (8.5-fold; 179.93 ± 9.82% of the normoxic control group, n = 5) and increasing Rab11B (15.5-fold) expression after 4 h of hypoxia. Similarly, Rab11A knockdown upregulated PAR1 expression under normoxia, and Rab11B knockdown downregulated PAR1 expression under both normoxic and hypoxic conditions. Cardiomyocytes knocked out of both Rab11A, and Rad11B lost the TRAP-induced PAR1 expression but still exhibited the early endosomal TRAP-induced PAR1 expression under hypoxia. Conclusions TRAP-mediated activation of PAR1 in cardiomyocytes did not alter the total PAR1 expression under normoxic conditions. Instead, it triggers a redistribution of PAR1 levels under normoxic and hypoxic conditions. TRAP reverses the hypoxia-inhibited PAR1 expression in cardiomyocytes by downregulating Rab11A expression and upregulating Rab11B expression. |
| first_indexed | 2024-04-09T23:11:22Z |
| format | Article |
| id | doaj.art-e901487394334cdd9f0a44cf41374529 |
| institution | Directory Open Access Journal |
| issn | 1471-2261 |
| language | English |
| last_indexed | 2024-04-09T23:11:22Z |
| publishDate | 2023-02-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cardiovascular Disorders |
| spelling | doaj.art-e901487394334cdd9f0a44cf413745292023-03-22T10:22:41ZengBMCBMC Cardiovascular Disorders1471-22612023-02-0123111610.1186/s12872-023-03118-wTRAP-induced PAR1 expression with its mechanism during AMI in a rat modelAni Wang0Xinyuan Gu1Chunyang Wang2Yanhui Li3Fuhong Deng4Jie Fang5Naxia Chen6Qifu Li7Lilong Tang8Department of Cardiology, The Fifth Affiliated Hospital of Sun Yat-Sen UniversityYuebei HospitalDepartment of Cardiology, The Fifth Affiliated Hospital of Sun Yat-Sen UniversityDivision of Cardiology, Tongji Hospital Affiliated to Huazhong Technology UniversityDepartment of Cardiology, The Fifth Affiliated Hospital of Sun Yat-Sen UniversityDivision of Cardiology, Xiangtan Central HospitalDepartment of Cardiology, The Fifth Affiliated Hospital of Sun Yat-Sen UniversityDepartment of Cardiology, The Fifth Affiliated Hospital of Sun Yat-Sen UniversityDepartment of Cardiology, The Fifth Affiliated Hospital of Sun Yat-Sen UniversityAbstract Background and objective Protease-activated receptor 1 (PAR1) is crucial in individuals with acute myocardial infarction (AMI). The continuous and prompt PAR1 activation mainly dependent on PAR1 trafficking is essential for the role of PAR1 during AMI in which cardiomyocytes are in hypoxia. However, the PAR1 trafficking in cardiomyocytes specially during the hypoxia is still unclear. Methods and result A rat AMI model was created. PAR1 activation with thrombin-receptor activated peptide (TRAP) had a transient effect on cardiac function in normal rats but persistent improvement in rats with AMI. Cardiomyocytes from neonatal rats were cultured in a normal CO2 incubator and a hypoxic modular incubator chamber. The cells were then subjected to western blot for the total protein expression and staining with fluorescent reagent and antibody for PAR1 localization. No change in total PAR1 expression following TRAP stimulation was observed; however, it led to increased PAR1 expression in the early endosomes in normoxic cells and decreased expression in the early endosomes in hypoxic cells. Under hypoxic conditions, TRAP restored the PAR1 expression on both cell and endosomal surfaces within an hour by decreasing Rab11A (8.5-fold; 179.93 ± 9.82% of the normoxic control group, n = 5) and increasing Rab11B (15.5-fold) expression after 4 h of hypoxia. Similarly, Rab11A knockdown upregulated PAR1 expression under normoxia, and Rab11B knockdown downregulated PAR1 expression under both normoxic and hypoxic conditions. Cardiomyocytes knocked out of both Rab11A, and Rad11B lost the TRAP-induced PAR1 expression but still exhibited the early endosomal TRAP-induced PAR1 expression under hypoxia. Conclusions TRAP-mediated activation of PAR1 in cardiomyocytes did not alter the total PAR1 expression under normoxic conditions. Instead, it triggers a redistribution of PAR1 levels under normoxic and hypoxic conditions. TRAP reverses the hypoxia-inhibited PAR1 expression in cardiomyocytes by downregulating Rab11A expression and upregulating Rab11B expression.https://doi.org/10.1186/s12872-023-03118-wPAR1Rab11CardiomyocytesHypoxiaCardiac functionAMI |
| spellingShingle | Ani Wang Xinyuan Gu Chunyang Wang Yanhui Li Fuhong Deng Jie Fang Naxia Chen Qifu Li Lilong Tang TRAP-induced PAR1 expression with its mechanism during AMI in a rat model BMC Cardiovascular Disorders PAR1 Rab11 Cardiomyocytes Hypoxia Cardiac function AMI |
| title | TRAP-induced PAR1 expression with its mechanism during AMI in a rat model |
| title_full | TRAP-induced PAR1 expression with its mechanism during AMI in a rat model |
| title_fullStr | TRAP-induced PAR1 expression with its mechanism during AMI in a rat model |
| title_full_unstemmed | TRAP-induced PAR1 expression with its mechanism during AMI in a rat model |
| title_short | TRAP-induced PAR1 expression with its mechanism during AMI in a rat model |
| title_sort | trap induced par1 expression with its mechanism during ami in a rat model |
| topic | PAR1 Rab11 Cardiomyocytes Hypoxia Cardiac function AMI |
| url | https://doi.org/10.1186/s12872-023-03118-w |
| work_keys_str_mv | AT aniwang trapinducedpar1expressionwithitsmechanismduringamiinaratmodel AT xinyuangu trapinducedpar1expressionwithitsmechanismduringamiinaratmodel AT chunyangwang trapinducedpar1expressionwithitsmechanismduringamiinaratmodel AT yanhuili trapinducedpar1expressionwithitsmechanismduringamiinaratmodel AT fuhongdeng trapinducedpar1expressionwithitsmechanismduringamiinaratmodel AT jiefang trapinducedpar1expressionwithitsmechanismduringamiinaratmodel AT naxiachen trapinducedpar1expressionwithitsmechanismduringamiinaratmodel AT qifuli trapinducedpar1expressionwithitsmechanismduringamiinaratmodel AT lilongtang trapinducedpar1expressionwithitsmechanismduringamiinaratmodel |