中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
5期
918-920
,共3页
李娜%王焱林%王成天%张宗泽%何祥虎
李娜%王焱林%王成天%張宗澤%何祥虎
리나%왕염림%왕성천%장종택%하상호
血红素氧合酶%心肌再灌注损伤%基因转染%腺病毒科%Toll样受体4
血紅素氧閤酶%心肌再灌註損傷%基因轉染%腺病毒科%Toll樣受體4
혈홍소양합매%심기재관주손상%기인전염%선병독과%Toll양수체4
Heme oxygenase%Myocardial reperfusion injury%Gene transfection%Adenoviridae%Toll-like receptor 4
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Toll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30 min、再灌注120 min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( Western blot)检测TLR4、肿瘤坏死因子-α(TNF-α)和核转录因子-κB (NF-κB)的蛋白表达水平.结果 rAAV-rHO-1组HO-1 mRNA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-α和NF-κB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 ±0.03、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TLR4表达水平,抑制炎症反应,从而减轻心肌IR损伤.
目的 觀察重組腺相關病毒(rAAV)介導大鼠血紅素氧閤酶-1(rHO-1)基因轉染對心肌缺血再灌註(IR)過程中Toll樣受體4(TLR4)錶達的影響.方法 構建攜帶rHO-1基因的rAAV 載體,併轉染大鼠心肌細胞.基因轉染後3箇月,用逆轉錄-聚閤酶鏈反應( RT-PCR)檢測HO-1mRNA錶達.採用結扎左冠狀動脈前降支30 min、再灌註120 min建立心肌IR模型.成模後測定心肌梗死麵積,光鏡下觀察心肌組織病理學改變,採用蛋白印跡法( Western blot)檢測TLR4、腫瘤壞死因子-α(TNF-α)和覈轉錄因子-κB (NF-κB)的蛋白錶達水平.結果 rAAV-rHO-1組HO-1 mRNA錶達(1.67±0.32)較IR組(0.82±0.09)增加,差異有統計學意義(P＜0.05),同時心肌梗死麵積較IR組減少(P＜0.05)、心肌組織病理學損傷較輕；rAAV-rHO-1組TLR4、TNF-α和NF-κB的蛋白錶達量分彆為(0.33 ±0.04、0.36±0.02、0.33±0.03),明顯低于IR組(0.45 ±0.03、0.42±0.05、0.46±0.07),差異有統計學意義(P均＜0.05).結論 rAAV介導的rHO-1基因轉染大鼠心肌細胞後,通過降低心肌TLR4錶達水平,抑製炎癥反應,從而減輕心肌IR損傷.
목적 관찰중조선상관병독(rAAV)개도대서혈홍소양합매-1(rHO-1)기인전염대심기결혈재관주(IR)과정중Toll양수체4(TLR4)표체적영향.방법 구건휴대rHO-1기인적rAAV 재체,병전염대서심기세포.기인전염후3개월,용역전록-취합매련반응( RT-PCR)검측HO-1mRNA표체.채용결찰좌관상동맥전강지30 min、재관주120 min건립심기IR모형.성모후측정심기경사면적,광경하관찰심기조직병이학개변,채용단백인적법( Western blot)검측TLR4、종류배사인자-α(TNF-α)화핵전록인자-κB (NF-κB)적단백표체수평.결과 rAAV-rHO-1조HO-1 mRNA표체(1.67±0.32)교IR조(0.82±0.09)증가,차이유통계학의의(P＜0.05),동시심기경사면적교IR조감소(P＜0.05)、심기조직병이학손상교경；rAAV-rHO-1조TLR4、TNF-α화NF-κB적단백표체량분별위(0.33 ±0.04、0.36±0.02、0.33±0.03),명현저우IR조(0.45 ±0.03、0.42±0.05、0.46±0.07),차이유통계학의의(P균＜0.05).결론 rAAV개도적rHO-1기인전염대서심기세포후,통과강저심기TLR4표체수평,억제염증반응,종이감경심기IR손상.
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 myocardial 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-α (TNF-α) and nuclear factor-κB (NF-κB) 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-α and NF-κB was significantly increased in IR group (0.45 ±0.03,0.42 ±0.05,and 0.46 ±0.07 respectively) 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 ＜0.05 ).The degree of damage to myocardial tissue was significantly severer in IR group than in SO group and rAAV-rHO-I group.Conclusion rAAV-mediated rHO-1 gene trans-fection may attenuate myocardial IR injury by reducing the TLR4 expression level and inhibiting the inflammatory response in rats.
