Effect and mechanism of HSPA1A on apoptosis of H9c2 cardiomyocytes in hypoxic environment
10.19405/j.cnki.issn1000-1492.2025.04.007
- VernacularTitle:HSPA1A对缺氧环境下H9c2心肌细胞凋亡损伤的影响及机制研究
- Author:
Xiaoling Li
1
;
Xiaolan Zhong
2
Author Information
1. Dept of Pharmacy，The Second Affiliated Hospital of Xinjiang Medical University，Urumqi 830063
2. Dept of Cardiovascular Medicine，The Second Affiliated Hospital of Xinjiang Medical University，Urumqi 830063
- Publication Type:Journal Article
- Keywords:
cardiomyocyte;
hypoxia;
heat shock protein 70 ku protein 1A;
inflammation;
apoptosis;
TNF recep- tor-associated factor 2;
nuclear factor κB
- From:
Acta Universitatis Medicinalis Anhui
2025;60(4):634-641
- CountryChina
- Language:Chinese
-
Abstract:
Objective :To investigate the effects of heat shock protein 70 ku protein 1A(HSPA1A) on H9c2 inflammation and apoptotic injury in rat cardiomyocytes under hypoxia, and to analyze its mechanism.
Methods:H9c2 cells were treated with normal oxygen(Nor) and hypoxia(Hyp), and the expression of HSPA1A was detected by RT-qPCR and Western blot. Normal H9c2 cells were divided into Nor group(normoxia culture), sh-HSPA1A+Nor group(cells transfected with HSPA1A shRNA, normoxia culture), Hyp group(hypoxia culture), sh-HSPA1A+Hyp group(cells transfected with HSPA1A SHRNA, hypoxia culture), the expression level of HSPA1A in H9c2 cells in each group was detected by RT-qPCR and Western blot, the morphology of H9c2 cells in each group was observed by inverted microscope, and the activity of H9c2 cells in each group was detected by CCK-8, the contents of inflammatory factors tumor necrosis factor-α(TNF-α), interleukin(IL)-6, IL-1β and the activities of myocardial injury markers lactate dehydrogenase(LDH), creatine kinase(CK) in the supernatant of H9c2 cells were detected by ELISA, apoptosis rate of H9c2 cells in each group was detected by Annexin V-FITC/PI double staining, the expression levels of TNF receptor-associated factor 2(TRAF2)/nuclear factor κB(NF-κB) pathway-related proteins in H9c2 cells of each group were detected by Western blot.
Results:Compared with Nor group, the relative expression of HSPA1A mRNA and protein in H9c2 cells in Hyp group after hypoxia induction was significantly up-regulated(P<0.05). Compared with Nor group, the number of H9c2 cells in Hyp group significantly decreased, some cells were wrinkled and disordered, the cell activity significantly decreased(P<0.05), the contents of TNF-α, IL-6, IL-1β and the activities of LDH and CK in supernatant significantly increased(P<0.05), the cell apoptosis rate significantly increased(P<0.05), the relative expressions of TRAF2 protein and the phosphorylation level of p65 and inhibitor of nuclear factor κB alpha(IκBα) were significantly up-regulated(P<0.05); compared with Hyp group, the morphology of H9c2 cells in sh-HSPA1A+Hyp group was improved, the cell arrangement was more dense, the cell activity significantly increased(P<0.05), the contents of TNF-α, IL-6 and IL-1β in supernatant decreased, the activities of LDH and CK significantly decreased(P<0.05), the apoptosis rate significantly decreased(P<0.05), the relative expressions of TRAF2 protein and the phosphorylation level of p65 and IκBα were also significantly down-regulated(P<0.05).
Conclusion :The expression of HSPA1A in rat cardiomyocytes H9c2 is increased by hypoxia, and inhibition of HSPA1A expression can improve the hypoxia-induced inflammatory response of H9c2 cells and reduce apoptosis damage, which may be related to the regulation of TRAF2/NF-κB pathway.
- Full text:202601260940121414HSPA1A对缺氧环境下H...胞凋亡损伤的影响及机制研究_李小玲.pdf
