摘要目的 探讨腺病毒重组血管生成素-1(adenovirus-delivered angiopoietin-1,Ad.Ang-1siRNA)对大鼠光气急性肺损伤基质金属蛋白酶-2,9(matrix metalloproteinase,MMP-2,9)及其组织抑制剂-1(tissue inhibitor of metalloproteinases,TIMP-1)表达的影响.方法 复制大鼠光气吸入性肺损伤动物模型.随机分为空气对照组(吸入与光气染毒组同等流量的空气)、空气空白Ad组(空气暴露后1h通过尾静脉注射1×108 pfu/ml Ad)、空气转染Ad/Ang1组(空气暴露后1h通过尾静脉注射1×108 pfu/ml Ad.Ang1 siRNA)、光气染毒组(吸入8.33 mg/L纯度为100％的光气5 min)、光气空白Ad组(给予同等剂量的光气吸入,光气染毒后1h通过尾静脉注射1×108 pfu/ml Ad)、光气转染Ad/Ang1组(给予同等剂量的光气吸入,光气染毒后1h通过尾静脉注射1×108 pfu/ml Ad.Ang-1 siRNA).染毒36 h后收集血清、支气管肺泡灌洗液(BALF)和肺组织.用双抗体夹心酶标免疫分析法(ELISA法)测定各组血清和BALF中Ang-1、MMP-2,9和TIMP-1蛋白含量;用反转录-聚合酶链式反应(RT-PCR)方法对肺组织Ang-1、MMP-2,9和TIMP-1mRNA表达量进行半定量研究;Western blot技术检测肺组织中MMP-2,9和TIMP-1蛋白表达量.结果 与空气对照组比较,光气染毒组和光气转染Ad/Ang1组血清、BALF中MMP-2,9蛋白含量及肺组织中MMP-2,9 mRNA和蛋白表达量明显升高,BALF中TIMP-1蛋白含量和肺组织中TIMP-1mRNA及蛋白表达量明显升高,差异有统计学意义(P＜0.01);血清中TIMP-1蛋白含量无明显变化,差异无统计学意义(P＞0.05).与光气染毒组比较,光气转染Ad/Ang1组血清、BALF中MMP-2,9蛋白含量和肺组织中MMP-2,9 mRNA及蛋白表达量明显减低,差异有统计学意义(P＜0.05),BALF中TIMP-1蛋白含量和肺组织中mRNA及蛋白表达量无明显变化.结论 Ad.Ang-1siRNA可能通过抑制MMP-2,9表达来有效地保护大鼠光气吸入性急性肺损伤,但对TIMP-1表达无明显影响.

abstractsObjective To investigate the effects of adenovirus-delivered angiopoietin-1 siRNA (Ad.Ang-1siRNA) on the expression of matrix metalloproteinase-2,9 (MMP-2,9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in rats with acute lung injury (ALI) induced by phosgene (Psg).Methods We first established a rat model of Psg-induced acute lung injury (ALl).The rats were randomly divided into 6 groups:air control group with exposure to air,air+adenovirus (air+Ad) group with caudal vein injection of 1×108 pfu/ml adenovirus 1 h after air exposure,air+Ad/Ang1 group with caudal vein injection of 1 ×108 pfu/ml Ad.Ang1siRNA 1 h after air exposure,Psg group with exposure to 8.33 mg/L Psg (purity 100％,of the same volume as the inhaled air in the air control group) for 5 min,Psg+Ad group with caudal vein injection of 1 ×108 pfu/ml adenovirus 1 h after exposure to the same dose of Psg,and Psg+Ad/Ang1 group with caudal vein injection of 1 × 108 pfu/ml Ad.Ang-1siRNA 1 h after exposure to the same dose of Psg.Serum,bronchoalveolar lavage fluid (BALF),and lung tissue were collected 36 h after exposure.The protein expression of Ang-1,MMP-2,9,and TIMP-1 in serum and BALF was determined by double-antibody sandwich ELISA.RT-PCR was used to determine the mRNA levels of Ang-1,MMP-2,9,and TIMP-1 in lung tissue.The protein expression of MMP-2,9 and TIMP-1 in lung tissue was determined by Western blot.Results A rat model of Psg-induced ALl was successfully established.The levels of MMP-2,9 in serum,BALF,and lung tissue were significantly increased in the Psg group and Psg+Ad/Ang1 group as compared with the control group (P＜0.01); no significant change was observed in serum TIMP-1 protein expression (P＞0.05); interestingly,TIMP-1 protein expression in BALF and lung tissue was significantly increased (P＜0.01).Compared with the Psg group,the Psg+Ad/Ang1 group showed a significant decrease in MMP-2,9 expression in BALF,serum,and lung tissue (P＜0.05),but no significant change in protein expression of TIMP-1 was discovered (P＞0.05).Conclusion Ad.Ang-1siRNA has a potential beneficial effect in rats with Psg-induced ALI through inhibition of MMP-2,9 expression,but has no significant effect on the expression of TIMP-1.