Molecular network mechanism in cerebral ischemia-reperfusion rats treated with human urine stem cells

Lang-Chun Zhang; Na Li; Ji-Lin Chen; Jie Sun; Min Xu; Wen-Qiang Liu; Zhong-Fu Zuo; Lan-Lan Shi; Ting-Hua Wang; Xiang-Yin Luo

doi:10.1016/j.heliyon.2024.e27508

Molecular network mechanism in cerebral ischemia-reperfusion rats treated with human urine stem cells

Heliyon. 2024 Mar 13;10(7):e27508. doi: 10.1016/j.heliyon.2024.e27508. eCollection 2024 Apr 15.

Authors

Lang-Chun Zhang^{1

2}, Na Li², Ji-Lin Chen², Jie Sun², Min Xu², Wen-Qiang Liu³, Zhong-Fu Zuo⁴, Lan-Lan Shi¹, Ting-Hua Wang¹, Xiang-Yin Luo¹

Affiliations

¹ Department of Neurosurgery, Xiang Ya Hospital of Central South University, Changsha, 410078, China.
² Animal Canter Department of Anatomy, Kunming Medical University, Kunming, 650500, China.
³ College of Basic Medicine, Jinzhou Medical University, Jinzhou, 121000, China.
⁴ Department of Anatomy, Jinzhou Medical University, Jinzhou, China.

Abstract

Objective: To explore the effect of human urine-derived stem cells (husc) in improving the neurological function of rats with cerebral ischemia-reperfusion (CIR), and report new molecular network by bioinformatics, combined with experiment validation.

Methods: After CIR model was established, and husc were transplanted into the lateral ventricle of rats，neurological severe score (NSS) andgene network analysis were performed. Firstly, we input the keywords "Cerebral reperfusion" and "human urine stem cells" into Genecard database and merged data with findings from PubMed so as to get their targets genes, and downloaded them to make Venny intersection plot. Then, Gene ontology (GO) analysis, kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and protein-protein interaction (PPI) were performed to construct molecular network of core genes. Lastly, the expressional level of core genes was validated via quantitative real-time polymerase chain reaction (qRT-PCR), and localized by immunofluorescence.

Results: Compared with the Sham group, the neurological function of CIR rats was significantly improved after the injection of husc into the lateral ventricle; at 14 days, P = 0.028, which was statistically significant. There were 258 overlapping genes between CIR and husc, and integrated with 252 genes screened from PubMed and CNKI. GO enrichment analysis were mainly involved neutrophil degranulation, neutrophil activation in immune response and platelet positive regulation of degranulation, Hemostasis, blood coagulation, coagulation, etc. KEGG pathway analysis was mainly involved in complement and coagulation cascades, ECM-receptor. Hub genes screened by Cytoscape consist ofCD44, ACTB, FN1, ITGB1, PLG, CASP3, ALB, HSP90AA1, EGF, GAPDH. Lastly, qRT-PCR results showed statistic significance (P < 0.05) in ALB, CD44 and EGF before and after treatment, and EGF immunostaining was localized in neuron of cortex.

Conclusion: husc transplantation showed a positive effect in improving neural function of CIR rats, and underlying mechanism is involved in CD44, ALB, and EGF network.

Keywords: Bioinformatics; Cerebral ischemia reperfusion; Human urine stem cells.