背景:Sirtuins(SIRT)家族在许多肾脏疾病的诊断和治疗中起着关键作用,但是没有关于肾移植急性排斥反应的研究报道。本研究旨在探讨SIRT家族改变特征在肾移植急性排斥反应中的诊断价值。
方法:我们首先使用HPA数据库探索肾组织中的SIRT家族表达谱;随后,我们通过GSEA富集分析探讨了肾移植急性排斥反应的潜在生物学功能和机制变化。Cibersort算法指定免疫细胞浸润水平,并利用相关性分析探讨SIRT家族与免疫细胞之间的相关性;接下来,我们使用“Logistic回归分析”和“列线图模型”构建了诊断模型,并使用校准曲线和ROC曲线评估诊断模型,用决策曲线(DCA)评价SIRT家族变化的临床诊断价值;最后,我们构建了大鼠肾移植急性排斥反应模型,通过检测血清尿素氮和肌酐水平评价大鼠肾功能。同时,通过转录组测序和RT-PCR初步验证了SIRT家族在肾组织中的表达水平。
结果:我们发现所有7个SIRT家族成员均在肾组织中定位并表达。富集分析结果显示,肾移植急性排斥反应过程中大量免疫相关的生物学功能和通路被激活,差异有统计学意义(p<0.05)。Cibersort算法显示10个免疫细胞浸润水平发生显著变化(p<0.05),而相关分析显示SIRT家族与免疫细胞之间有很强的联系(p<0.05)。我们使用七个SIRT家族构建了急性排斥反应的诊断模型,ROC曲线(AUC=0.71)和校准曲线证明了其良好的诊断价值,DCA曲线也证明了SIRT家族在临床决策中的作用。接下来,我们再次证明了SIRT家族在ABMR和TCMR中的良好诊断性能,ROC曲线:AUC=0.64,AUC=0.81。最后,在肾移植急性排斥反应的大鼠模型中,我们发现,与Syn组相比,Allo组大鼠的肾功能(BUN和肌酐)明显受损(P<0.05)。同时,通过转录组分析和RT-PCR分析,我们发现,除SIRT1外,其余SIRT家族成员肾组织均有显著改变(P<0.05)。
结论:SIRT家族在肾移植急性排斥反应过程中有显著变化,SIRT家族可能成为减轻肾移植急性排斥反应的潜在治疗靶点。
BACKGROUND: The Sirtuins (SIRT) family plays a key role in the diagnosis and treatment of many renal diseases, but no studies have been reported in acute rejection of kidney transplantation. The aim of this study was to explore the diagnostic value of SIRT family change characteristics in acute rejection of kidney transplantation.
METHODS: We first explored the SIRT family expression profile in renal tissues using the HPA database; subsequently, we explored the potential biological functions and mechanistic changes during acute rejection of kidney transplantation by GSEA enrichment analysis. The Cibersort algorithm specifies the level of immune cell infiltration and explores the correlation between the SIRT family and immune cells using correlation analysis; Next, we constructed a diagnostic model using \"Logistic regression analysis\" and \"Nomogram model\", and evaluated the diagnostic model using calibration curves and ROC curves, and the decision curve (DCA) was used to evaluate the clinical diagnostic value of SIRT family changes; Finally, we constructed a model of acute rejection of rat kidney transplantation, and assessed rat kidney function by detecting the levels of urea nitrogen and creatinine in serum. Meanwhile, the expression level of SIRT family in kidney tissues was initially verified by transcriptome sequencing and RT-PCR.
RESULTS: We found that all seven SIRT family members were located and expressed in renal tissues. The results of enrichment analysis revealed that a large number of immune-related biological functions and pathways are activated during acute rejection of kidney transplantation, the difference was statistically significant (p < 0.05). The Cibersort algorithm revealed significant changes in the level of infiltration of 10 immune cells (p < 0.05), while correlation analysis revealed a strong link between the SIRT family and immune cells (p < 0.05). We constructed a diagnostic model for acute rejection using seven SIRT families, and the ROC curves(AUC = 0.71)and calibration curves proved their good diagnostic value, and the DCA curves also proved the role of SIRT families in clinical decision-making. Next, we again demonstrated the good diagnostic performance of the SIRT family in ABMR and TCMR, respectively(ROC curves:AUC = 0.64,AUC = 0.81). Finally, in a rat model of acute rejection of kidney transplantation, we found that renal function (BUN and creatinine) was significantly impaired in rats in the Allo group compared to rats in the Syn group (P < 0.05). Meanwhile, by transcriptome analysis and RT-PCR assay, we found that, except for SIRT1, the remaining SIRT family members were significantly changed in kidney tissues (P < 0.05).
CONCLUSIONS: The SIRT family has significant changes during acute rejection in kidney transplantation, and the SIRT family may be able to serve as a potential therapeutic target for alleviating acute rejection in kidney transplantation.