Abstract:Tripartite motif 8 (TRIM8) is a member of the TRIM protein family that has been found to be implicated in cardiovascular disease. However, the role of TRIM8 in myocardial ischemia/reperfusion (I/R) has not been investigated. We aimed to explore the effect of TRIM8 on cardiomyocyte H9c2 cells exposed to hypoxia/reoxygenation (H/R). We found that TRIM8 expression was markedly upregulated in H9c2 cells after stimulation with H/R. Gain- and loss-of-function assays proved that TRIM8 knockdown improved cell viabilit… Show more
“…It was reported that knockdown of TRIM8 suppressed H/R-generated oxidative stress and apoptosis in cardiac H9C2 cells, which was achieved through PI3K/Akt signaling pathway. 18 Notably, blockage of TRIM8 enhanced phosphorylation of PI3K/Akt in si-TRIM8 group versus si-NC group, which represented that PI3K/Akt signaling pathway was activated by TRIM8 inhibition ( Figure 5A and B ). Figure 5 PI3K/Akt signaling pathway was involved in TRIM8 inhibition-regulated H/R injury in HK-2 cells.…”
Section: Resultsmentioning
confidence: 90%
“… 16 Downregulation of TRIM8 was confirmed to mitigate damage by relieving oxidative stress and apoptosis in oxygen–glucose deprivation/re-oxygenation-induced injury in neurons, 17 as well as in H/R-induced injury in cardiac H9C2 cells. 18 The results suggested that the protective effect of TRIM8 inhibition in neurons was associated with reinforcement of the AMPK/Nrf2/ARE pathway, which was an important antioxidant pathway. In cardiac H9C2 cells, its protective effect was connected with the activation of PI3K/Akt pathway.…”
Section: Discussionmentioning
confidence: 97%
“… 17 Moreover, the decrease of TRIM8 expression resulted in the activated PI3K/Akt signaling pathway to protect H9c2 cells against H/R-induced oxidative stress and apoptosis. 18 Despite the intensive research of TRIM8 on various pathological processes, the role of TRIM8 in renal I/R injury remains unclarified. According to these findings, we speculated that TRIM8 could regulate renal I/R injury.…”
Background
Acute kidney injury (AKI) emerges as an acute and critical disease. Tripartite motif 8 (TRIM8), one number of the TRIM protein family, is proved to participate in ischemia/reperfusion (I/R) injury. However, whether TRIM8 is involved in renal I/R injury and the associated mechanisms are currently unclear.
Purpose
This study aimed to investigate the precise role of TRIM8 and relevant mechanisms in renal I/R injury.
Materials and Methods
In this study, human renal proximal tubular epithelial cells (HK-2 cells) underwent 12 hours of hypoxia and 2 h, 3 h or 4 h of reoxygenation to establish an in vitro hypoxia/reoxygenation (H/R) model. The siRNAs specific to TRIM8 (si-TRIM8) were transfected into HK-2 cells to knockdown TRIM8. The cell H/R model included various groups including Control, H/R, H/R+DMSO, H/R+NAC, si-NC+H/R, si-TRIM8+H/R and si-TRIM8+LY294002+H/R. The cell viability and levels of reactive oxygen species (ROS), hydrogen peroxide (H
2
O
2
), mRNA, apoptotic proteins, pyroptosis-related proteins and PI3K/AKT pathway-associated proteins were assessed.
Results
In vitro, realtime-quantitative PCR and western-blot analysis showed that the mRNA and protein expression of TRIM8 were obviously upregulated after H/R treatment in HK-2 cells. Compared with the H/R model group, knockdown of TRIM8 significantly increased cell viability and reduced the levels of ROS, H
2
O
2
, apoptotic proteins (Cleaved caspasebase-3 and BAX) and pyroptosis-related proteins (NLRP3, ASC, Caspase-1, Caspase-11, IL-1β and GSDMD-N). Western-blot analysis also authenticated that PI3K/AKT pathway was activated after TRIM8 inhibition. The application of 5 mM N-acetyl-cysteine, one highly efficient ROS inhibitor, significantly suppressed the expression of apoptotic proteins and pyroptosis-related proteins. Moreover, the combined treatment of TRIM8 knockdown and LY294002 reversed the effects of inhibiting oxidative stress.
Conclusion
Knockdown of TRIM8 can alleviate H/R-induced oxidative stress by triggering the PI3K/AKT pathway, thus attenuating pyropyosis and apoptosis in vitro.
“…It was reported that knockdown of TRIM8 suppressed H/R-generated oxidative stress and apoptosis in cardiac H9C2 cells, which was achieved through PI3K/Akt signaling pathway. 18 Notably, blockage of TRIM8 enhanced phosphorylation of PI3K/Akt in si-TRIM8 group versus si-NC group, which represented that PI3K/Akt signaling pathway was activated by TRIM8 inhibition ( Figure 5A and B ). Figure 5 PI3K/Akt signaling pathway was involved in TRIM8 inhibition-regulated H/R injury in HK-2 cells.…”
Section: Resultsmentioning
confidence: 90%
“… 16 Downregulation of TRIM8 was confirmed to mitigate damage by relieving oxidative stress and apoptosis in oxygen–glucose deprivation/re-oxygenation-induced injury in neurons, 17 as well as in H/R-induced injury in cardiac H9C2 cells. 18 The results suggested that the protective effect of TRIM8 inhibition in neurons was associated with reinforcement of the AMPK/Nrf2/ARE pathway, which was an important antioxidant pathway. In cardiac H9C2 cells, its protective effect was connected with the activation of PI3K/Akt pathway.…”
Section: Discussionmentioning
confidence: 97%
“… 17 Moreover, the decrease of TRIM8 expression resulted in the activated PI3K/Akt signaling pathway to protect H9c2 cells against H/R-induced oxidative stress and apoptosis. 18 Despite the intensive research of TRIM8 on various pathological processes, the role of TRIM8 in renal I/R injury remains unclarified. According to these findings, we speculated that TRIM8 could regulate renal I/R injury.…”
Background
Acute kidney injury (AKI) emerges as an acute and critical disease. Tripartite motif 8 (TRIM8), one number of the TRIM protein family, is proved to participate in ischemia/reperfusion (I/R) injury. However, whether TRIM8 is involved in renal I/R injury and the associated mechanisms are currently unclear.
Purpose
This study aimed to investigate the precise role of TRIM8 and relevant mechanisms in renal I/R injury.
Materials and Methods
In this study, human renal proximal tubular epithelial cells (HK-2 cells) underwent 12 hours of hypoxia and 2 h, 3 h or 4 h of reoxygenation to establish an in vitro hypoxia/reoxygenation (H/R) model. The siRNAs specific to TRIM8 (si-TRIM8) were transfected into HK-2 cells to knockdown TRIM8. The cell H/R model included various groups including Control, H/R, H/R+DMSO, H/R+NAC, si-NC+H/R, si-TRIM8+H/R and si-TRIM8+LY294002+H/R. The cell viability and levels of reactive oxygen species (ROS), hydrogen peroxide (H
2
O
2
), mRNA, apoptotic proteins, pyroptosis-related proteins and PI3K/AKT pathway-associated proteins were assessed.
Results
In vitro, realtime-quantitative PCR and western-blot analysis showed that the mRNA and protein expression of TRIM8 were obviously upregulated after H/R treatment in HK-2 cells. Compared with the H/R model group, knockdown of TRIM8 significantly increased cell viability and reduced the levels of ROS, H
2
O
2
, apoptotic proteins (Cleaved caspasebase-3 and BAX) and pyroptosis-related proteins (NLRP3, ASC, Caspase-1, Caspase-11, IL-1β and GSDMD-N). Western-blot analysis also authenticated that PI3K/AKT pathway was activated after TRIM8 inhibition. The application of 5 mM N-acetyl-cysteine, one highly efficient ROS inhibitor, significantly suppressed the expression of apoptotic proteins and pyroptosis-related proteins. Moreover, the combined treatment of TRIM8 knockdown and LY294002 reversed the effects of inhibiting oxidative stress.
Conclusion
Knockdown of TRIM8 can alleviate H/R-induced oxidative stress by triggering the PI3K/AKT pathway, thus attenuating pyropyosis and apoptosis in vitro.
“…Notably, the Wnt signalling pathway is a classical neuron development signalling pathway [ 42 ]. Meanwhile, TRIM8 [ 43 ] and RPSA [ 44 ] are also an important regulators of the PI3K-AKT-mTOR signalling pathway, which is a developmental disease-related signalling pathway [ 40 ]. LHX2 is a regulator of neural differentiation [ 45 , 46 ].…”
De novo variants (DNVs) are critical to the treatment of neurodevelopmental disorders (NDDs). However, effectively identifying candidate genes in small cohorts is challenging in most NDDs because of high genetic heterogeneity. We hypothesised that integrating DNVs from multiple NDDs with genetic similarity can significantly increase the possibility of prioritising the candidate gene. We catalogued 66,186 coding DNVs in 50,028 individuals with nine types of NDDs in cohorts with sizes spanning from 118 to 31,260 from Gene4Denovo database to validate this hypothesis. Interestingly, we found that integrated DNVs can effectively increase the number of prioritised candidate genes for each disorder. We identified 654 candidate genes including 481 shared candidate genes carrying putative functional variants in at least two disorders. Notably, 13.51% (65/481) of shared candidate genes were prioritised only via integrated analysis including 44.62% (29/65) genes validated in recent large cohort studies. Moreover, we estimated that more novel candidate genes will be prioritised with the increase in cohort size, in particular for some disorders with high putative functional DNVs per individual. In conclusion, integrated DNVs may increase the power of prioritising candidate genes, which is important for NDDs with small cohort size.
“…At the same time, in H/R-stimulated H9c2 cells TRIM8 silencing causes a marked increase in Bcl-2 expression and the activation of the PI3K/Akt signaling pathway, which is well documented in being associated with a decrease in myocardial ischemic injury. PI3K/Akt activation inhibits cardiomyocyte apoptosis induced by hypoxia, and it protects against I/R injury [ 63 ].…”
The superfamily of TRIM (TRIpartite Motif-containing) proteins is one of the largest groups of E3 ubiquitin ligases. Among them, interest in TRIM8 has greatly increased in recent years. In this review, we analyze the regulation of TRIM8 gene expression and how it is involved in many cell reactions in response to different stimuli such as genotoxic stress and attacks by viruses or bacteria, playing a central role in the immune response and orchestrating various fundamental biological processes such as cell survival, carcinogenesis, autophagy, apoptosis, differentiation and inflammation. Moreover, we show how TRIM8 functions are not limited to ubiquitination, and contrasting data highlight its role either as an oncogene or as a tumor suppressor gene, acting as a “double-edged weapon”. This is linked to its involvement in the selective regulation of three pivotal cellular signaling pathways: the p53 tumor suppressor, NF-κB and JAK-STAT pathways. Lastly, we describe how TRIM8 dysfunctions are linked to inflammatory processes, autoimmune disorders, rare developmental and cardiovascular diseases, ischemia, intellectual disability and cancer.
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