血红素氧合酶-1在HO-1/Luc小鼠原位肝脏移植模型中的表达

Heme oxygenuse-1 expression in the orthotopic liver transplantation model of HO-1/Lue mice

目的 应用活体生物萤光成像技术（BLT）无创定量检测血红素氧合酶-1（HO-1）在活体动物肝脏移植模型中的时空表达.方法 建立小鼠原位肝脏移植模型（HO-1/Luc转基因小鼠8只,wildtype小鼠40只）,使用活体生物萤光成像技术连续检测HO-1在移植术后HO-1/Luc转基因小鼠中的转录.应用逆转录-聚合酶链反应（RT-PCR）检测HO-1 mRNA水平,免疫组织化学方法 （IHC）行HO-1的表达定位.结果 移植肝脏发出的萤光信号最早于移植后1 h被检测到（P〈0.05）,随后信号不断增强在6 h达到峰值.与移植术前比较,除0、48 h外信号差异均有统计学意义（P〈0.05）.移植后48 h信号衰减至基础水平.与移植术前比较,RT-PCR方法 测得HO-1 mRNA于0～9 h均显著升高（P〈0.05）,其中于3 h达到峰值,12 h降至基础水平.IHC证实肝脏细胞是移植后HO-1表达上调的主要部位.结论 活体生物萤光成像技术可实时定量检测肝脏移植后HO-1的表达.

Objective To investigate the feasibility of in vivo bioluminescence imaging （BLI） to non-invasively quantify the spatiotemporal expression of heine oxygenase-1 （HO-1） after fiver transplantation （LT） in living animals. Methods We established the mice LT model （Hol-luc n=8, wildtype n=40）.Luciferase activity was measured by BLI as an index of HO-1 transcription in transgenic reporter mice （Hol-lue）. HO-1 mRNA leveLs were measured in post-transplant livers of mice. Immunohistochemistry （IHC） was used to locate the HO-1 protein. Results Bioluminescent signals from liver were first detected at 1st h after transplantation （P〈0.05）, and reached the peak at 6th h, then bioluminescent activity declined and returned to the baseline value at 48th h after transplantation. Signals were higher than baseline at various time points but 0 and 48 h （P〈0.05）. HO-1 mRNA expression was increased from 0 to 9 h after LT （P〈 0.05）, reached the peak at 3rd h, and decreased to baseline at 12th h. IHC confirmed that HO-1 was de-rived from hepatocytes. Conclusion In vivo BLI allows a quantitative assessment of HO-1 expression after liver transplantation in living animals.