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A Rat Model of Middle Cerebral Artery Occlusion/Reperfusion Without Damaging the Anatomical Structure of Cerebral Vessels
Published on: May 17, 2024
Receptor-interacting protein kinase 3-mediated programmed cell necrosis in rats subjected to focal cerebral
Yanru Dong1, Cuifen Bao2, Jingwei Yu1
1Department of Histology and Embryology, Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China.
Abstract:
In the current study, the activation of tumor necrosis factor-α receptor 1 (TNFR1) and receptor-interacting protein kinase 3 (RIP3) were investigated following cerebral ischemia-reperfusion injury (CIRI). Healthy SD rats were randomly divided into 3 groups: Sham operation group, model group and inhibitor group. The model group and inhibitor group were further divided into 4 subgroups of 6, 12, 24 and 72 h following CIRI. Using right middle cerebral artery embolization, the CIRI model was generated. To confirm that the CIRI model was established, neurological scores, TTC staining and brain water content measurements were conducted. Immunohistochemistry and western blotting were conducted to investigate the expression of TNFR1 and RIP3 in the cerebral cortex. It was observed that nerve cell necrosis occurred following 6 h of CIRI. The appearance of necrotic cells was gradually increased with increasing CIRI duration. TNFR1 and RIP3 were positively expressed following 6 h of CIRI. With increasing durations of CIRI, the protein expression levels of TNFR1 and RIP3 were significantly increased. Pre‑administration with Z-VAD-FMK (zVAD) significantly increased the protein level of RIP3, however, had no effect on the levels of TNFR1, and was accompanied by a reduction in necrosis. In conclusion, RIP3‑mediated cell necrosis was enhanced by caspase blockade zVAD and the function of zVAD was independent of TNFR1 signaling following IR.
Insights
Caspase blockade enhances receptor-interacting protein kinase 3 (RIP3)-mediated cell necrosis following cerebral ischemia-reperfusion injury (CIRI). This effect is independent of tumor necrosis factor-α receptor 1 (TNFR1) signaling.
Area of Science:
- Neuroscience
- Cell Biology
- Pathology
Background:
- Cerebral ischemia-reperfusion injury (CIRI) is a critical condition leading to neuronal damage.
- Tumor necrosis factor-α receptor 1 (TNFR1) and receptor-interacting protein kinase 3 (RIP3) are implicated in cell death pathways.
- Understanding the roles of TNFR1 and RIP3 in CIRI is crucial for developing therapeutic strategies.
Purpose of the Study:
- To investigate the activation of TNFR1 and RIP3 in a rat model of CIRI.
- To determine the effect of caspase blockade using Z-VAD-FMK (zVAD) on RIP3 and TNFR1 expression and neuronal necrosis.
- To elucidate the relationship between zVAD, RIP3, TNFR1, and cell death in CIRI.
Main Methods:
- A rat model of CIRI was established using middle cerebral artery embolization.
- Neurological scores, TTC staining, and brain water content were used to confirm the CIRI model.
- Immunohistochemistry and Western blotting were employed to assess TNFR1 and RIP3 expression.
Main Results:
- Neuronal necrosis was observed starting at 6 hours post-CIRI and increased with duration.
- Both TNFR1 and RIP3 expression were upregulated following CIRI, with levels increasing over time.
- zVAD treatment increased RIP3 protein levels and reduced necrosis but did not affect TNFR1 levels.
Conclusions:
- RIP3-mediated cell necrosis is enhanced by caspase blockade with zVAD following CIRI.
- The protective function of zVAD in this context is independent of TNFR1 signaling.
- These findings highlight RIP3's role in CIRI-induced necrosis and suggest potential therapeutic targets.
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