Objective To investigate the changes of differentially expressed genes before and after in the liver xenotransplantation, and they were screened and verified. Methods Four cases of xenograft splenic fossa heterotopic auxiliary liver transplantation were performed with a -1，3- galactoside transferase gene knockout（GTKO） mini-pig as donor and Tibetan macaque as recipient. Appropriate preoperative liver specimens were harvested from the donor liver at the back table procedure，and postoperative liver specimens were harvested by biopsy at 48 h afteropening the blood flow of the graft. High throughput chip analysis of miRNAs and mRNAs was performed on the transplanted liver specimens. The key differential expression of miRNAs and mRNAs were screened，and real-time quantitative PCR and Western Blot tests were performed. Results There were 165 miRNAs differentially expressed after surgery，and 112 of them were up-regulated and 53 were down-regulated. Twelve differentially expressed miRNAs，such as mi-23b，were screened and verified. In addition to miR-150 and miR-126-3p，the expression changes of other miRNAs were basically consistent with the results of miRNAs chip. Compared with preoperative data，2 035 mRNAs showed significant differential expression after operation，922 of them were up-regulated and 1 113 were down-regulated，5 differentially expressed genes which were closely related to liver transplantation, such as IGFBP6，CDK2AP1，TNFAIP6，FⅦ and NKG2D，were screened at mRNA and protein levels. Except for NKG2D，the changes in the protein level of the other 4 genes were basically consistent with the changes in the mRNA level（P ＜ 0.05），but the changes in the CDK2AP1 protein were small，and there was no statistical difference （P ＝ 0.073）. Conclusion The expression of miRNAs and mRNAs in transplanted liver after xenotransplantation has greatly changed. These differentially expressed genes may be related to immune injury and coagulation dysfunction.

目的 探讨异种移植手术前后移植肝内差异表达基因的变化，并且进行筛选及验证。 方法 以 α-1，3- 半乳糖苷转移酶基因敲除（GTKO）小型猪为供体、藏酋猴为受体，完成 4 例异种脾 窝辅助性肝移植手术。于供肝修整时切取适量术前标本，于移植肝开放血流后 48 小时以肝穿刺活检获得 术后标本。对手术前后的移植肝标本进行 miRNAs 和 mRNAs 的高通量芯片分析，筛选出关键的差异表达 miRNAs 和 mRNAs，并进行实时定量 PCR 及 Western Blot 实验验证。结果 与术前相比，有 165 个 miRNAs 在术后出现差异表达，其中上调分子 112 个，下调分子 53 个。筛选并验证 mi-23b 等 12 个出现显著差异 表达的 miRNAs，除 miR-150 和 miR-126-3p 外，其他 miRNAs 的表达变化情况与 miRNAs 芯片结果基本相 符。与术前相比，有 2 035 个 mRNAs 在术后出现显著差异表达，其中 922 个表达上调、1 113 个表达下调， 筛选 IGFBP6、CDK2AP1、TNFAIP6、FⅦ、NKG2D 等 5 个生物学功能与肝脏移植密切相关的差异表达基因 进行 mRNA 和蛋白水平验证。除 NKG2D 外，其他 4 个基因的蛋白水平表达变化与 mRNA 水平变化基本一致 （均 P ＜ 0.05），但 CDK2AP1 的蛋白变化幅度较小，差异无统计学差异（P ＝ 0.073）。结论 异种肝移植术 后移植肝的 miRNAs 和 mRNAs 表达出现较大变化，这些差异表达基因可能与免疫损伤及凝血功能障碍有关。