双 mTORC1/2 抑制剂 AZD2014 可抑制大鼠肝移植后的急性排斥反应

目的 在同种异基因大鼠肝移植模型中验证AZD2014是否具有抑制肝移植术后急性排斥反应的作用。方法 采用Kamada 提出的“二袖套”法建立Lewis→BN 同种异基因大鼠肝移植急性排斥反应模型，随机分成对照组和AZD2014组，各4只。AZD2014组腹腔内注射AZD2014药物，5 mg/kg，1次/d；对照组腹腔内注射药物溶剂2.5 mL/kg，1次/d。不同时间点取外周血检测肝功能（丙氨酸氨基转移酶、天门冬氨酸氨基转移酶和总胆红素）。记录生存时间，进行生存分析。移植肝脏行免疫组化检测CD3和Foxp3的表达水平，评估T淋巴细胞和Treg淋巴细胞浸润的程度；移植肝脏行HE染色，采用Banff方案评估肝移植术后排斥反应的严重程度。结果 对照组在术后14 d内有3/4 的大鼠死亡，而AZD2014组在术后14 d内无大鼠死亡，AZD2014组与对照组相比生存时间明显延长（χ2=4.213，P=0.040）。对照组血清中ALT、AST和TBIL进行性升高，上述指标均高于同时间AZD2014组（P<0.05）。病理检查显示对照组移植肝内排斥反应明显重于AZD2014组（排斥指数P<0.01），对照组中T淋巴细胞（CD3阳性）浸润相较于AZD2014组更为严重（P<0.01），而Treg细胞（Foxp3阳性）明显少于AZD2014组（P<0.01）。结论 双mTORC1/2抑制剂AZD2014可以有效地抑制同种异基因大鼠肝移植术后的急性排斥反应。

The dual mTORC1/2 inhibitor AZD2014 inhibits acute graft rejection in a rat liver transplantation model

Objective To investigate the inhibitory effect of AZD2014, a dual mTORC1/2 inhibitor, against acute graft rejection in a rat model of allogeneic liver transplantation. Methods Liver transplantation from Lewis rat to recipient BN rat (a donor-recipient combination that was prone to induce acute graft rejection) was performed using Kamada's two-cuff technique. The recipient BN rats were randomized into 2 groups for treatment with daily intraperitoneal injection of AZD2014 (5 mg/kg, n=4) or vehicle (2.5 mL/kg, n=4) for 14 consecutive days, starting from the first day after the transplantation. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBIL) levels of the rats were measured 3 days before and at 1, 3, 5, 7, 10, and 14 days after the transplantation, and the survival time of the rats within 14 days were recorded. Immunohistochemical staining was used to examine the expressions of CD3 and Foxp3 in the liver graft, and acute graft rejection was assessed using HE staining based on the Banff schema. Results Three rats in the control group died within 14 days after the surgery, while no death occurred in the AZD2014 group, demonstrating a significantly longer survival time of the rats in AZD2014 group (χ2=4.213, P=0.04). Serum ALT, AST and TBIL levels in the control group increased progressively after the surgery and were all significantly higher than those in AZD2014 group at the same time point (P<0.05). Pathological examination revealed significantly worse liver graft rejection in the control group than in AZD2014 group based on assessment of the rejection index (P<0.01); the rats in the control group showed more serious T lymphocyte infiltration and significantly fewer Treg cells in the liver graft than those in AZD2014 group (P<0.01). Conclusions AZD2014 can effectively inhibit acute graft rejection in rats with allogeneic liver transplantation.