摘要
目的观察重组腺相关病毒（rAAV）介导大鼠血红素氧合酶-1（rHO-1）基因转染对心肌缺血再灌注（IR）过程中T011样受体4（TLR4）表达的影响。方法构建携带rHO-1基因的rAAV载体，并转染大鼠心肌细胞。基因转染后3个月，用逆转录-聚合酶链反应（RT—PCR）检测HO-1mRNA表达。采用结扎左冠状动脉前降支30min、再灌注120min建立心肌IR模型。成模后测定心肌梗死面积，光镜下观察心肌组织病理学改变，采用蛋白印迹法（Westernblot）检测TLR4、肿瘤坏死因子-a（TNF-a）和核转录因子-KB（NF—KB）的蛋白表达水平。结果rAAV—rHO-1组HO-1mRNA表达（1．67±0．32）较IR组（0．82±0．09）增加，差异有统计学意义（P〈0．05），同时心肌梗死面积较IR组减少（P〈0．05）、心肌组织病理学损伤较轻；rAAV—rHO-1组TLR4、TNF-a和NF—KB的蛋白表达量分别为（0．33±0．04、0．36±0．02、0．33±0．03），明显低于IR组（0．45±0．05、0．42±0．05、0．46±0．07），差异有统计学意义（P均〈0．05）。结论rAAV介导的rHO-1基因转染大鼠心肌细胞后，通过降低心肌TLRd表达水平，抑制炎症反应，从而减轻心肌IR损伤。
Objective To investigate the effect of rat hemeoxygenase-1 （ rHO-1 ） gene carried by recombinant adeno-associated virus （rAAV） on expression of Toll-like receptor 4 （TLR4） during myocar-dial ischemia-reperfusion （IR） injury in rats. Methods rHO-1 carry gene rAAV carrier was constructed, then transfected into rat myocardial cells. After 3 months, the expression of HO-1 mRNA in the injected myocardium was detected by using reverse transcription-polymerase chain reaction （RT-PCR）. After the successful reproduction of the myocardial IR model, the animals were killed and their hearts were harvested for determination of myocardial infarct size and pathological changes in myocardial tissue. The expression of TLR4, tumor necrosis factor-a （TNF-ct） and nuclear factor-KB （NF-KB） was detected by using Western blotting. Results The expression of HO-1 mRNA was significantly higher, and the infarct size was significantly smaller in rAAV-rHO-1 group than in group IR （both P 〈 0. 05）. The expression of TLR4, TNF-a and NF-KB was significantly increased in IR group （0. 45±0. 03, 0.42±0. 05, and 0.46±0. 07 respec-tively） and rAAV-rHO-1 group （0. 33±0. 04, 0. 36±0. 02, and 0. 33±0. 03 respectively） as compared with rAAV-rHO-1 group （ all P 〈 O. 05 ）. The degree of damage to myocardial tissue was significantly severer in IR group than in SO group and rAAV-rHO-1 group. Conclusion rAAV-mediated rHO-1 gene trans- fection may attenuate myocardial IR injury by reducing the TLR4 expression level and inhibiting the inflam- matory response in rats.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第5期918-920,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30471658）
湖北省自然科学基金资助项目（2004ABAl62）
