Exploring the Mechanism of RIP3 in Thalamic Hemorrhage Based on Animal Models and Transcriptome Sequencing

Methods

The 24 male C57BL/6 mice (8-10 weeks/20-25 g) were equally divided into wild type (WT) group, RIP3 knockout (KO) group, TH wild type (TH-WT) group, and TH RIP3 knockout (TH-KO) group. First, TH model was constructed, and the behavioral and morphological characteristics of 4 groups of mice were analyzed. Next, RNA sequencing was performed on brain tissue samples obtained from 4 groups of mice, and the resulting sequencing data were integrated into subsequent analyses.

Thereafter, differentially expressed genes (DEGs) were identified between WT and KO groups, as well as between TH-WT and TH-KO groups, referred to as DEGs1 and DEGs2. Then, DEGs1 was removed from DEGs2 to obtain candidate genes. The Molecular COmplex Detection (MCODE) plugin was subsequently utilized to filter key genes.

Eventually, expression and enrichment analysis were performed to further explore potential mechanisms between TH and RIP3. Results: Mice with TH displayed abnormal gait, elevated pain thresholds, heightened anxiety levels, impaired learning and memory function, more severe hemorrhages, larger areas of brain tissue edema, and severe damage to the myelin sheath. Nevertheless, knockdown of RIP3 partially mitigated these detrimental effects.

Furthermore, We identified 956 candidate genes by removing 490 DEGs1 in 1,011 DEGs2. And 6 key genes: Pdyn, Gal, Tac1, Hcrt, Pomc and Drd1 were ultimately identified, which exhibited low expression levels in TH-KO groups. Key genes were enriched in signaling pathways related to immune response and biological process, such as toll like receptor signaling pathway and ribosome pathway.

Conclusion: Mice with TH exhibited compromised behavioral and morphological characteristics, and knockdown of RIP3 alleviated these detrimental effects. The identification of Pdyn, Gal, Tac1, Hcrt, Pomc and Drd1 as RIP3-related key genes in TH established a foundation for further investigations into the underlying mechanisms of TH and RIP3.