血红素氧合酶-1基因转染对移植小肠的保护作用

Protective effects of heme oxygenase-1 gene transfection on small bowel grafts

目的观察重组腺相关病毒血红素氧合酶-1（HO-1）基因转染对移植小肠的保护作用。方法建立大鼠同种异位小肠移植模型，实验分为2组：实验组（n=34）：供体术前腹腔注射载有HO-1基因的腺相关病毒1．0×10^12个／ml，对照组（n=34）：供体术前腹腔注射腺相关病毒1．0×10^12个／ml，在实验组和对照组中检测小肠移植术后小肠组织中HO-1的活性、HO．1mRNA含量及细胞凋亡，免疫组织化学检测HO-1蛋白的表达，观察小肠组织病理学变化和受体生存时间。结果实验组平均生存时间为（6．80±1．56）d，对照组平均生存时间为（5．80±1．28）d，两者之间差异无统计学意义（P〉0．05），小肠移植术后24、48h和5d移植小肠实验组HO-1活性分别为2．11±0．04、1．90±0．04和1．56±0．05，均强于对照组（1．71±0．07、1．56±0．06和1．38±0．08，P〈0．05）；各组移植肠缺血再灌注损伤程度以术后24h最重，实验组缺血再灌注损伤程度轻于对照组：术后24h和48h实验组HO-1mRNA表达水平分别为1．12±0．08和0．76±0．10，均高于对照组（0．69±0．05和0．38±0．08，P〈0．05），术后5d HO-1mRNA表达水平两组比较差异无统计学意义（P〉0．05）；术后各时段移植小肠细胞凋亡数实验组低于对照组，差异有统计学意义（P〈0．05），生存时间差异无统计学意义（P〉0．05）。结论HO-1基因转染可降低移植小肠缺血再灌注损伤程度。

Objective To evaluate the expression of heme oxygenase-1 （ HO-1 ） gene in small in- testines and its action on alleviating the ischemia-raperfusion injury in transplanted small bowel. Methods Ischemia-reperfulion injury model of smaU bowel transplantation was established. Wistar rats were divided into two groups： experimental group （n = 34） ： rAAV-HO-1 was delivered to doner rats via intraperitoneal injection before transplantation; control group （n = 34） ： rAAV was delivered to doner rats via intraperitoneal injection before transplantation. The activity of HO-1 in the tissue of transplanted intestine was detected, histological changes observed and apoptosis detected. The expression of HO-1 protein and mRNA in the transplanted small bowel tissue was detected by using immunohistochemistry and reverse transcription- polymerase chain reaction （RT-PCR） respectively. Results The average survival time in experimental group and control group was （6. 80 ± 1.56） and （5.80 ± 1.28 ） days respectively （ P 〉 0. 05 ）. The post- operative HO-1 actitivity in the allografts in experimental group at 24 h, 48 h and 5 day was 2. 11 ± 0.04, 1.90 ±0. 04 and 1.56 ± 0. 05, respectively, which was higher than that in control group （ 1.71 ± 0. 07, 1.56 ±0.06 and 1.38 ±0. 08 ,P 〈0.05）. All groups showed the most serious ischemia-reperfusion injury 24 h after reperfusion, and the damage was milder in experimental group than that in control group. The post-operative expression of HO-1 mRNA in the allografts in experimental group at 24 h and 48 h was 1.12 ± 0. 08 and 0. 76 ± 0. 10, respectively, which was higher than that in control group （0. 69 ± 0. 05 and 0. 38 ±0. 08）, and there was no significant difference in the expression of HO-1 mRNA between experimental group and control group 5 days after transplantation （ P 〉 0. 05 ）, The post-operative HO-1 actitivity in the allografts in experimental group at 24 h, 48 h and 5 day was 2. 11± 0.04, 1.90 ± 0.04 and 1.56 ± 0. 05, respectively, which was higher than that in control group （ 1.71 ±0. 07, 1.56 ± 0.06 and. 1.38 ± 0.08 ） （ P 〈 0. 05 ）. There were less apoptosis cells in experimental group than control group （ P 〈 0. 05 ）, and there was no significant difference in survival time. Conclusion HO-1 gene transfection can alleviste the ischamia-reperfusion injury of transplanted small bowel.