Department of Pathology and Institute of Oncology, The School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian, China.
Department of Pathology, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, Fujian, China.
Cytokine. 2021 Dec;148:155689. doi: 10.1016/j.cyto.2021.155689. Epub 2021 Sep 8.
Prevention of acute rejection is the key of the success of liver transplantation. However, there are no specific indicators available for prediction of acute rejection after liver transplantation. MicroRNAs (miRNAs) are highly conserved and small noncoding RNA molecules that can be detected in peripheral blood. Here, we evaluated the potential of circulating miRNAs to serve as noninvasive biomarkers for acute rejection after liver transplantation in rats.
The liver grafts retrieved from Lewis rats were orthotopically transplanted into BN rats or Lewis rats in the acute rejection and immune tolerance group respectively, and the BN rats in the immune intervention group was intraperitoneally injected with transforming growth factor-β1 overexpressed immature dendritic cells to suppress acute rejection before orthotopically transplanted with livers from Lewis rats. MiRNAs profiling studies were used to determine the regulation of circulating miRNAs in plasma samples of rats. Candidate miRNA was verified by quantitative reverse transcriptase polymerase chain reaction. Furthermore, the relationship between candidate miRNA and acute rejection was also evaluated.
Microarray analysis revealed that miR-199a-3p was the mostly differentially regulated miRNAs in plasma samples among the three groups. The plasmid PCDH-CMV-EGFP-hTGF-β1 was identified by PCR and DNA sequencing, and successfully expressed in imDCs. There were differences in the expression of miR-199a-3p in the liver tissues of the AR group on the 3rd, 7th and 10th day after liver transplantation (all p < 0.01). With time, the RAI score increased gradually, and the difference of miR-199a-3p expression gradually increased (rs = 0.92, p < 0.001), suggesting that it may be related to acute rejection. The expression of miR-199a-3p in the serum of the AR and TGF-β1-imDCs groups gradually increased, reaching a peak at day 7 and then decreasing. There was positive relationship between the expression of miR-199a-3p and RAIs within 7 days post operation. (rs = 0.942, p < 0.05).
miR-199a-3p might be an early warning marker for acute rejection after liver transplantation in rats.
预防急性排斥反应是肝移植成功的关键。然而，目前尚无预测肝移植后急性排斥反应的特异性指标。微小 RNA（miRNA）是高度保守的小非编码 RNA 分子，可在外周血中检测到。在这里，我们评估了循环 miRNA 作为大鼠肝移植后急性排斥反应无创生物标志物的潜力。
从 Lewis 大鼠中取出的肝移植物分别在急性排斥和免疫耐受组中原位移植到 BN 大鼠或 Lewis 大鼠中，在 BN 大鼠中，在原位移植 Lewis 大鼠的肝之前，通过腹腔注射过表达转化生长因子-β1 的未成熟树突状细胞来抑制急性排斥反应。使用 miRNA 谱研究确定大鼠血浆样本中循环 miRNA 的调节。通过定量逆转录聚合酶链反应验证候选 miRNA。此外，还评估了候选 miRNA 与急性排斥反应之间的关系。
微阵列分析显示，miR-199a-3p 是三组血浆样本中差异最大的调节 miRNA。通过 PCR 和 DNA 测序鉴定出质粒 PCDH-CMV-EGFP-hTGF-β1，并在 imDC 中成功表达。肝移植后第 3、7 和 10 天，AR 组肝组织中 miR-199a-3p 的表达存在差异（均 p < 0.01）。随着时间的推移，RAI 评分逐渐升高，miR-199a-3p 表达的差异逐渐增大（rs=0.92，p < 0.001），提示其可能与急性排斥反应有关。AR 组和 TGF-β1-imDC 组血清中 miR-199a-3p 的表达逐渐升高，在第 7 天达到峰值，然后下降。miR-199a-3p 的表达与术后 7 天内的 RAI 呈正相关（rs=0.942，p < 0.05）。
miR-199a-3p 可能是大鼠肝移植后急性排斥反应的早期预警标志物。
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