Chen Qiangxing, Zhou Rou, Zhang Yingcai, Zhu Shuguang, Xiao Cuicui, Gong Jiao, Li Kun, Tang Hui, Sun Chong, Zhang Jian
Liver Transplantation Center, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China; Guang First People's Hospital, The Second Affiliated Hospital, South China University of Technology, Guangzhou, Guangdong 510180, China.
Guang First People's Hospital, The Second Affiliated Hospital, South China University of Technology, Guangzhou, Guangdong 510180, China.
Transpl Immunol. 2018 Dec;51:21-29. doi: 10.1016/j.trim.2018.08.004. Epub 2018 Aug 6.
Studies have reported that bone marrow mesenchymal stem cells (BMSCs) play an important role in immune regulation after organ transplantation. Some of the BMSC-mediated regulatory mechanisms are related to PD-L1 expression. However, the regulatory mechanism of PD-L1 expression is not fully understood. In this experiment, we confirmed the regulatory role of BMSCs in the rejection of liver transplantation and explored the possible mechanism by which PD-L1 expression is regulated in BMSCs. An orthotopic liver transplantation model in rats was established based on the "double-cuff technique". Third-generation BMSCs were injected into the rejection model rats via portal vein. At the same time, sera were obtained from rats in the allograft, isograft and sham-operated groups. The sera from these groups were separately added to BMSCs complete medium to partially mimic the in vivo environment in which BMSCs are exposed. After predicting and analysing microRNAs that regulate PD-L1 expression, we examined the microRNA and PD-L1 expression in BMSCs of the three groups cultured in the conditioned media. Moreover, a luciferase reporter assay was used to confirm the interaction between PD-L1 and microRNA. The study found that the liver function and liver graft histopathology of the BMSC-treated group were better than those of the allograft group. The Kaplan-Meier survival curve analysis showed that the median survival time (MST) of the BMSC-treated group was longer than that of the allograft group. Bioinformatics prediction and analysis showed that miR-17-5p is highly likely to regulate PD-L1. Compared with the other two groups of cells, BMSCs cultured with serum from allograft models showed higher PD-L1 expression, but lower miR-17-5p expression. Pearson correlation analysis showed that miR-17-5p and PD-L1 expression was negatively correlated, and further luciferase reporter assays confirmed that miR-17-5p interacted directly with the 3´-untranslated region (UTR) of PD-L1 mRNA. The results of this study demonstrate that BMSCs can attenuate liver allograft rejection and provide us with the idea that BMSCs may further upregulate PD-L1 expression by downregulating miR-17-5p expression, thereby attenuating liver allograft rejection.
研究报告称,骨髓间充质干细胞(BMSCs)在器官移植后的免疫调节中发挥重要作用。一些BMSC介导的调节机制与PD-L1表达有关。然而,PD-L1表达的调节机制尚未完全明确。在本实验中,我们证实了BMSCs在肝移植排斥反应中的调节作用,并探讨了BMSCs中PD-L1表达的可能调节机制。基于“双袖套技术”建立大鼠原位肝移植模型。将第三代BMSCs经门静脉注射到排斥模型大鼠体内。同时,从同种异体移植组、同基因移植组和假手术组大鼠中获取血清。将这些组的血清分别添加到BMSCs完全培养基中,以部分模拟BMSCs所处的体内环境。在预测和分析调节PD-L1表达的微小RNA后,我们检测了在条件培养基中培养的三组BMSCs中的微小RNA和PD-L1表达。此外,使用荧光素酶报告基因检测来确认PD-L1与微小RNA之间的相互作用。研究发现,BMSC治疗组的肝功能和肝移植组织病理学均优于同种异体移植组。Kaplan-Meier生存曲线分析表明,BMSC治疗组的中位生存时间(MST)长于同种异体移植组。生物信息学预测和分析表明,miR-17-5p极有可能调节PD-L1。与其他两组细胞相比,用同种异体移植模型血清培养的BMSCs显示出更高的PD-L1表达,但miR-17-5p表达较低。Pearson相关性分析表明,miR-17-5p与PD-L1表达呈负相关,进一步的荧光素酶报告基因检测证实miR-17-5p与PD-L1 mRNA的3´-非翻译区(UTR)直接相互作用。本研究结果表明,BMSCs可减轻肝移植排斥反应,并为我们提供了这样一个思路,即BMSCs可能通过下调miR-17-5p表达进一步上调PD-L1表达,从而减轻肝移植排斥反应。