Abd El-Aziz Ahlam, El-Desouky Mohamed Ali, Shafei Ayman, Elnakib Mostafa, Abdelmoniem Amr Mohamed
Department of Chemistry, Biochemistry Division, Faculty of Science, Cairo University, Egypt.
Military Medical Academy, Faculty of Medicine, Modern University for Technology and Information, Cairo, Egypt.
Biochem Biophys Rep. 2021 Jan 27;25:100911. doi: 10.1016/j.bbrep.2021.100911. eCollection 2021 Mar.
The regulation by immune checkpoint is able to prevent excessive tissue damage caused by ischemia reperfusion (I/R); therefore, the study aims to investigate the behavior of phosphoprotein associated with glycosphingolipid-enriched microdomains 1 (PAG1) mRNA, miR-1206 and small nucleolar RNA host gene 14 (SNHG14) during I/R and intake of pentoxifylline (PTX) as a protective drug. The relative expression level of PAG1/miR-1206/SNHG14 was determined by qRT-PCR. Cardiac tissue levels of cytotoxic T-lymphocyte associated antigen 4 (CTLA4) and PAG1 protein expression were determined by ELISA technique. The regulatory T cells achieved by the flow cytometry. The results found that the relative expression of SNHG14 was significantly upregulated in I/R, but suppressed in PTX treated groups with enhancement of the relative expression level of miR-1206. The gene and protein expression of PAG1 were downregulated with effective doses of PTX. The results showed that (30 and 40 mg/kg bwt) PTX dose suppressed the CTLA4 development significantly. The mean of the regulatory T cell in PTX protective groups is significantly reduced at (p < 0.001) in a comparison with I/R group. Spearman's correlation analysis revealed a significant negative correlation between SNHG14 and miR-1206, but a significant positive correlation between SNHG14 and PAG1 in I/R heart tissue. The results indicated that miR-1206 and SNHG14 can be used as biomarkers with perfect sensitivity and specificity. Using PTX reduced cardiac tissue damage. SNHG14 and miR-1206 can be used as a diagnostic tool in I/R.
免疫检查点调节能够预防缺血再灌注（I/R）引起的过度组织损伤；因此，本研究旨在调查富含糖鞘脂微结构域1（PAG1）mRNA、miR-1206和小核仁RNA宿主基因14（SNHG14）在I/R期间的行为以及作为保护药物的己酮可可碱（PTX）的摄入情况。通过qRT-PCR测定PAG1/miR-1206/SNHG14的相对表达水平。采用ELISA技术测定心脏组织中细胞毒性T淋巴细胞相关抗原4（CTLA4）水平和PAG1蛋白表达。通过流式细胞术检测调节性T细胞。结果发现，SNHG14的相对表达在I/R中显著上调，但在PTX治疗组中受到抑制，同时miR-1206的相对表达水平增强。PAG1的基因和蛋白表达在有效剂量的PTX作用下下调。结果显示，（30和40mg/kg体重）PTX剂量显著抑制了CTLA4的发展。与I/R组相比，PTX保护组中调节性T细胞的平均值在（p<0.001）时显著降低。Spearman相关性分析显示，在I/R心脏组织中，SNHG14与miR-1206之间存在显著负相关，但SNHG14与PAG1之间存在显著正相关。结果表明，miR-1206和SNHG14可作为具有完美敏感性和特异性的生物标志物。使用PTX可减少心脏组织损伤。SNHG14和miR-1206可作为I/R的诊断工具。
