抗血管内皮生长因子抗体对转化生长因子β和多西环素诱导的兔胸膜粘连的影响

Effect of anti-vascular endothelial growth factor antibody on pleurodesis induced by transforming growth factor-β or doxycycline in rabbits

目的探讨抗血管内皮生长因子抗体(anti-VEGF antibody)对转化生长因子β(TGF-β)或多西环素(doxycyc line,Doxy)所诱导的胸腔积液增多和胸膜粘连作用的影响。方法28只新西兰大白兔随机分4组:TGF-β对照组、TGF-β抗体组、Doxy对照组和Doxy抗体组,每组7只。在放置胸导管后,TGF-β对照组经胸腔内注入TGF-β5.0μg;TGF-β抗体组在胸腔内注入TGF-β前24 h先予以静脉注射抗血管内皮生长因子抗体10 mg/kg。Doxy对照组胸腔内注射Doxy 10 mg/kg;Doxy抗体组在胸腔内注入Doxy前24 h先予以静脉注射抗血管内皮生长因子抗体10 mg/kg。在TGF-β或Doxy注射后的24、48、72 h分别收集胸腔积液并计量,测定胸腔积液中白细胞数、蛋白及乳酸脱氢酶(LDH)含量。2周后杀死实验兔并作胸膜粘连评分,通过免疫组化染色对胸膜组织中血管新生程度做定量分析。结果4组实验兔所产生的胸腔积液量,胸腔积液中白细胞数、蛋白量及LDH水平差异无统计学意义(P>0.05)。胸膜粘连评分TGF-β对照组为(7.7±0.8)分,TGF-β抗体组为(4.4±2.4)分;Doxy对照组为(6.0±1.7)分,Doxy抗体组为(2.0±0.9)分。胸膜组织中新生血管密度TGF-β对照组为(4.9±0.4)%,TGF-β抗体组为(2.9±0.7)%;Doxy对照组为(6.9±2.2)%,Doxy抗体组为(2.2±0.9)%。两抗体干预组的胸膜粘连评分和血管新生程度均显著低于对照组(P<0.05)。结论抗血管内皮生长因子抗体显著抑制了TGF-β或Doxy所诱导的胸膜粘连,提示VEGF和血管新生可能在胸膜固定术中起重要作用。

Objective The intrapleural injection of transforming growth factor-β （TGF-β） or doxycycline produces excellent pleurodesis in rabbit models. However, the intrapleural injection of these agents induces large pleural effusion which is possibly related to vascular endothelial growth factor （VEGF）. This study investigated whether anti-VEGF antibody has any effect on the fluid production or the pleurodesis induced by TGF-β or doxycycline in rabbits. Methods Two groups of New Zealand white rabbits （ 7 each） were given TGF-β 5.0 μg intrapleurally. The TGF-β treatment group received anti-VEGF antibody 10 mg/kg intravenously 24 h before TGF-β injection and the TGF-β control group received no antibody. Another two groups of New Zealand white rabbits （7 each） were given doxycycline 10 mg/kg intraplettrally after chest tube placement. The doxycycline treatment group received 10 mg/kg anti-VEGF antibody intravenously 24 h before doxycycline injection and the doxycycline control group received no Anti-VEGF antibody. The rabbits were sacrificed at 2 weeks and the pleurodesis score was graded macroscopically on a 1-8 scale. The degree of angiogenesis in pleural tissues was assessed by immunohistochemical staining for factor Ⅷ which was assessed by computer-assisted digital analysis. Results The administration of anti- VEGF antibodies had no effect on pleural fluid volume or the characteristics of the fluid. The mean pleurodesis score of TGF-β control group （ 7. 7 ± 0. 8） was significantly （ P 〈 0. 05 ） higher than that of the antibody pre-treatment TGF-β group （4. 4 ± 2. 4）. The mean pleurodesis score of the doxycycline control group （6. 0 ± 1.7） was significantly （ P 〈 0. 05 ） higher than that of the antibody pre-treatment doxycycline group （2. 0 ± 0. 9 ）. The administration of the anti-VEGF antibody also reduced the angiogenesis. The percentage of pleural tissue demonstrating angiogenesis in the TGF-β control group （4. 9 ± O. 4） % was significantly （P 〈 0. 05 ） higher than that of the antibody treatment TGF-β group （2.9 ± 0. 7 ） %. The percentage of pleural angiogenesis in the doxycycline control group （ 6. 9 ± 2.2 ） % was significantly （ P 〈 0. 05 ） higher than the antibody pre-treatment doxycycline group （2.2 ± 0. 9） %. Conclusions Anti-VEGF antibody significantly inhibits the pleurodesis induced by doxycycline or TGF-β. This observation suggests that VEGF and angiogenesis play a pivotal role in the production of pleurodesis.