课题背景：排斥反应是影响器官移植后人及供体长期存活的重要因素。研究表明，内皮细胞和组织细胞中血红素加氧酶1等保护基因的表达可以明显延长移植物存活时间。这些保护基因表达上调不仅阻滞前炎症因子的产生，而且能够抑制内皮细胞凋亡。

应用要点：实验以肾作为移植器官，以BN和Lewis大鼠作为供、受体，成功建立了双侧供肾大鼠同种异体肾移植急性排斥反应模型，通过氯高铁血红素诱导组织细胞中保护基因血红素加氧酶1的表达，分析血红素加氧酶1在肾脏移植中的抗免疫排斥和抗凋亡作用。

偏倚或不足：血红素加氧酶1在肾移植急性排斥反应的病理生理过程中以及移植免疫应答过程中起重要作用，但还不能完全替代常规的环孢素A等强力免疫抑制剂。

摘要

背景：研究表明，血红素加氧酶1系统及其代谢产物可通过多条途径发挥对器官移植物的保护作用，显著延长移植物生存时间。
目的：实验拟进一步评估氯高铁血红素诱导产生的内源性血红素加氧酶1对大鼠肾移植急性排斥反应的防护作用。
设计、时间及地点：随机对照动物实验，于2007-05/08在解放军第三军医大学动物实验中心完成。
材料：以27只BN大鼠为供体（54个供肾），54只雄性Lewis大鼠为受体，建立双侧供肾大鼠原位肾移植模型。
方法：以硬膜外导管为支架行供肾静脉与受体肾静脉端端吻合，供肾动脉带腹主脉瓣与受体腹主动脉行端侧吻合，供肾输尿管膀胱瓣与受者膀胱吻合。Lewis受鼠按随机数字表法分为3组（n=18）：肾移植模型组术后不予免疫抑制治疗；氯高铁血红素诱导组供鼠于术前1 d腹腔注射氯高铁血红素50 mg/kg，受体于术后腹腔注射氯高铁血红素 50 mg/（kg?d）至处死；环孢素A组术后予环孢素A灌胃5 mg/（kg?d）。
主要观察指标：各组分别于肾移植术后第1，5，7 天处死6只受鼠，观察肾组织病理改变、血红素加氧酶1蛋白表达及其对急性排斥反应的影响，同时采集血标本测血肌酐含量。
结果：①氯高铁血红素可促进大鼠肾脏血红素加氧酶1蛋白表达水平(P < 0.05)。②移植术后肾移植模型组、氯高铁血红素诱导组的血红素加氧酶1表达水平高于环孢素A组(P < 0.05~0.01)，且随着排斥反应的加重，表达逐渐增强；环孢素A组在移植术后血红素加氧酶1表达也有所增强，但明显低于氯高铁血红素诱导组(P < 0.01)，排斥反应相对较轻。③氯高铁血红素诱导组大鼠肾组织病理学表现较肾移植模型组明显减轻，但比环孢素A组重。④氯高铁血红素诱导组血肌酐水平低于肾移植模型组(P < 0.05)，高于环孢素A组(P < 0.05)。
结论：血红素加氧酶1在大鼠同种异体肾移植抗排斥反应、保护移植器官中起到了一定的作用，但其作用远远不及环孢素A等强力免疫抑制剂。
关键词：肾移植；氯化血红素；血红素氧化酶；疾病模型，动物；器官移植

Protective effect of hemin induced endogenous heme oxygenase-1 on acute rejection following renal transplantation in rats

Abstract

BACKGROUND: Heme oxygenase-1 and its metabolic product have a protective effect on organ graft and can significantly prolong live time of the graft.
OBJECTIVE: To investigate the protective effect of heme oxygenase-1 induced by hemin on acute rejection in rat models after renal transplantation.
DESIGN, TIME AND SETTING: A randomized control animal experiment was performed at the Animal Experimental Center of Third Military Area Command of Chinese PLA from May to August 2007.
MATERIALS: Totally 27 adult Brown-Norway rats (54 kidneys) as donors and 54 male Lewis rats as receptors were used to establish rat models of in situ renal transplantation.
METHODS: Epidural catheter was used as a stent to perform end-to-end anastomosis of renal vein between donors and receptors, end-to-side anastomosis of donor abdominal aorta valve and receptor abdominal aorta, and donor kidney tubal bladder valve and receptor bladder anastomosis. Lewis rats were equally and randomly divided into three groups. Rats in the renal transplantation group did not receive immunosuppressive therapy after transplantation. Hemin (50 mg/kg) was intraperitoneally injected into receptor rats in the hemin group one day before surgery. Hemin (50 mg/kg) was daily intraperitoneally injected into receptor rats after surgery. Ciclosporin A 5 mg/kg was given to rats in the ciclosporin A group by lavage after surgery.
MAIN OUTCOME MEASURES: Six rats were sacrificed at days 1, 5 and 7 after renal transplantation in each group. Pathological changes in renal tissues, heme oxygenase-1 protein expression and the heme oxygenase-1 effects on acute rejection were observed. Blood samples were collected to measure serum creatinine content.
RESULTS: Hemin could promote heme oxygenase-1 protein expression in rat kidney (P < 0.05). Heme oxygenase-1 levels were higher in the renal transplantation and hemin groups than in the ciclosporin A group (P < 0.05-0.01). With the severe of rejection, heme oxygenase-1 levels increased. Heme oxygenase-1 levels increased in the ciclosporin A group after transplantation, but significantly decreased in the hemin group (P < 0.01). Rejection was mild. Renal histopathological changes were fewer in the hemin group compared with the renal transplantation group, but more compared with the ciclosporin A group. Serum creatinine levels were lower in the hemin group than in the renal transplantation group (P < 0.05), but higher than in the ciclosporin A group (P < 0.05).
CONCLUSION: Heme oxygenase-1 has a protective effect on acute rejection in rat allograft renal transplantation, but its effect is significantly inferior to ciclosporin A.

Li SD, Jin FS, Li QS, Zhu FQ, Nie ZL, Huo WQ, Bi G, Ma Q. Protective effect of hemin induced endogenous heme oxygenase-1 on acute rejection following renal transplantation in rats.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(18):3427-3431 [www.zglckf.com/zglckf/ejournal/upfiles/08-18/18k-3427(ps).pdf]