中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
2期
184-185
,共2页
宿华威%李建国%孙铁为%孙世波%申宝忠
宿華威%李建國%孫鐵為%孫世波%申寶忠
숙화위%리건국%손철위%손세파%신보충
肝脏移植%血红素氧合酶-1%缺血%再灌注损伤
肝髒移植%血紅素氧閤酶-1%缺血%再灌註損傷
간장이식%혈홍소양합매-1%결혈%재관주손상
Liver transplantation%Heine oxygenase-1%Ischemia%Reperfusion injury
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.
目的 應用活體生物螢光成像技術(BLT)無創定量檢測血紅素氧閤酶-1(HO-1)在活體動物肝髒移植模型中的時空錶達.方法 建立小鼠原位肝髒移植模型(HO-1/Luc轉基因小鼠8隻,wildtype小鼠40隻),使用活體生物螢光成像技術連續檢測HO-1在移植術後HO-1/Luc轉基因小鼠中的轉錄.應用逆轉錄-聚閤酶鏈反應(RT-PCR)檢測HO-1 mRNA水平,免疫組織化學方法 (IHC)行HO-1的錶達定位.結果 移植肝髒髮齣的螢光信號最早于移植後1 h被檢測到(P<0.05),隨後信號不斷增彊在6 h達到峰值.與移植術前比較,除0、48 h外信號差異均有統計學意義(P<0.05).移植後48 h信號衰減至基礎水平.與移植術前比較,RT-PCR方法 測得HO-1 mRNA于0～9 h均顯著升高(P<0.05),其中于3 h達到峰值,12 h降至基礎水平.IHC證實肝髒細胞是移植後HO-1錶達上調的主要部位.結論 活體生物螢光成像技術可實時定量檢測肝髒移植後HO-1的錶達.
목적 응용활체생물형광성상기술(BLT)무창정량검측혈홍소양합매-1(HO-1)재활체동물간장이식모형중적시공표체.방법 건립소서원위간장이식모형(HO-1/Luc전기인소서8지,wildtype소서40지),사용활체생물형광성상기술련속검측HO-1재이식술후HO-1/Luc전기인소서중적전록.응용역전록-취합매련반응(RT-PCR)검측HO-1 mRNA수평,면역조직화학방법 (IHC)행HO-1적표체정위.결과 이식간장발출적형광신호최조우이식후1 h피검측도(P<0.05),수후신호불단증강재6 h체도봉치.여이식술전비교,제0、48 h외신호차이균유통계학의의(P<0.05).이식후48 h신호쇠감지기출수평.여이식술전비교,RT-PCR방법 측득HO-1 mRNA우0～9 h균현저승고(P<0.05),기중우3 h체도봉치,12 h강지기출수평.IHC증실간장세포시이식후HO-1표체상조적주요부위.결론 활체생물형광성상기술가실시정량검측간장이식후HO-1적표체.
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.
