Sun Bo, Gao Junwei, Shi Weifeng, Guo Yankun, Fan Junwei, Zhang Jigang, Li Xiaoyu, Liu Gaolin
Department of Pharmacy, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China.
Shanghai Center for Drug Evaluation and Inspection, Shanghai, PR China.
Pharmacogenomics. 2019 Apr;20(6):447-456. doi: 10.2217/pgs-2018-0198. Epub 2019 Feb 25.
This study aimed to investigate the effect of and mechanism involved in the SNP on cyclosporine metabolism and outcomes of liver transplantation (LT). The genotype, IL-17 expression, postoperative outcome and cyclosporine concentration were reviewed in 106 LT recipients. The functional relevance of rs2275913 was evaluated by luciferase assay. Furthermore, L02 cells were treated with IL-17 recombinant protein or/and pregnane X receptor (PXR) knockdown lentiviruses, then the expression of PXR, CYP3A4, CYP3A5 and IL-17R were detected by PCR and western blotting. The significant distribution difference at locus G-197A was confirmed between patients with and without rejection (p = 0.035). Patients with acute rejection showed higher IL-17 level than those without rejection. Cyclosporine concentration was associated with the different genotype (p < 0.05). Luciferase assay revealed that 197G genotype had higher luciferase activity than that in 197A genotype (p = 0.009). Furthermore, IL-17 recombinant protein remarkably promoted the expressions of PXR, CYP3A4 and CYP3A5 (p < 0.01), but not IL-17R. PXR knockdown significantly inhibited the mRNA levels of CYP3A4 and CYP3A5 but not IL-17R (p < 0.01), while IL-17 recombinant protein had no influence on the expressions of CYP3A4 and CYP3A5 when PXR was downregulated. This study revealed the possible association of IL-17 G-197A with cyclosporine metabolism and transplant rejection after LT, which might be partly related to the upregulations of CYP3A4/5 dependent on PXR.
本研究旨在探讨单核苷酸多态性（SNP）对环孢素代谢及肝移植（LT）结局的影响及其机制。回顾性分析了106例LT受者的基因型、白细胞介素-17（IL-17）表达、术后结局及环孢素浓度。通过荧光素酶报告基因检测评估rs2275913的功能相关性。此外，用IL-17重组蛋白或/和孕烷X受体（PXR）敲低慢病毒处理L02细胞，然后通过聚合酶链反应（PCR）和蛋白质免疫印迹法检测PXR、细胞色素P450 3A4（CYP3A4）、细胞色素P450 3A5（CYP3A5）和IL-17受体（IL-17R）的表达。证实有排斥反应和无排斥反应患者在G-197A位点存在显著的分布差异（p = 0.035）。急性排斥反应患者的IL-17水平高于无排斥反应患者。环孢素浓度与不同基因型相关（p < 0.05）。荧光素酶报告基因检测显示，197G基因型的荧光素酶活性高于197A基因型（p = 0.009）。此外，IL-17重组蛋白显著促进PXR、CYP3A4和CYP3A5的表达（p < 0.01），但对IL-17R无影响。PXR敲低显著抑制CYP3A4和CYP3A5的mRNA水平，但对IL-17R无影响（p < 0.01），而当PXR下调时，IL-17重组蛋白对CYP3A4和CYP3A5的表达无影响。本研究揭示了IL-17 G-197A与LT后环孢素代谢及移植排斥反应可能存在关联，这可能部分与依赖PXR的CYP3A4/5上调有关。
