博舒替尼对内毒素血症小鼠急性肺损伤的影响

Effects of bosutinib on acute lung injury in mice with endotoxemia

目的:评价博舒替尼对内毒素血症小鼠急性肺损伤的影响。方法:清洁级健康雄性C57BL/6小鼠60只,8~12周龄,体重20~25 g,采用随机数字表法分为4组(n=15):对照组(C组)、博舒替尼组(B组)、内毒素血症组(LPS组)和博舒替尼+内毒素血症组(B+LPS组)。采用腹腔注射LPS 10 mg/kg的方法制备内毒素性急性肺损伤模型。B+LPS组于造模前0.5 h、B组于相应时点,尾静脉注射博舒替尼5 mg/kg。于造模后24 h时,处死小鼠,观察肺组织病理学结果,并行肺损伤评分,计算肺系数(LI)、肺湿重/干重(W/D)比值;行伊文思蓝染色,检测肺组织伊文思蓝含量;采用Western blot法检测血管内皮钙黏蛋白(VE-cadherin)、血管细胞粘附分子1(VCAM-1)、血管内皮钙黏蛋白(VE-cadherin)、磷酸化核转录因子κB p65(p-NF-κB p65)和磷酸化核因子κB抑制蛋白α(pIκB-α)的表达,采用RT-PCR法检测IL-1β、IL-6、TNF-α的mRNA表达。结果:与C组比较,LPS组LI、肺W/D比值、肺组织伊文思蓝含量和肺损伤评分升高,肺组织IL-1βmRNA、TNF-αmRNA、IL-6 mRNA、VCAM-1、p-NF-κB p65和pIKB-α表达上调,VE-cadherin表达下调(P<0.05),B组上述指标比较差异无统计学意义(P>0.05)。与LPS组比较,B+LPS组LI、肺W/D比值、肺组织伊文思蓝含量和肺损伤评分降低,肺组织IL-1βmRNA、TNF-αmRNA、IL-6 mRNA、VCAM-1、p-NF-κB p65和pIKB-α表达下调,VE-cadherin表达上调(P<0.05)。结论:博舒替尼可减轻内毒素血症小鼠急性肺损伤。

Objective To evaluate the effects of bosutinib on acute lung injury in mice with endotoxemia.Methods Sixty clean-grade healthy male C57BL/6 mice,aged 8-12 weeks,weighing 20-25 g,were divided into 4 groups(n=15 each)using a random number table method:control group(group C),bosutinib group(group B),endotoxemia group(group lipopolysaccharide[LPS])and bosutinib plus endotoxemia group(group B+LPS).Septic acute lung injury model was developed by intraperitoneal injection of LPS.Bosutinib 5 mg/kg was injected via the tail vein at 0.5 h before establishing the model in group B+LPS and at the corresponding time point in group B.At 24 h after developing the model,the mice were sacrificed for microscopic examination of the pathological results of lung tissues which were scored for calculation of the lung coefficient(LI)and wet/dry lung weight(W/D)ratio,and for determination of the content of Evans blue in lung tissues(by Evans blue staining),expression of vascular endothelial cadherin(VE-cadherin),vascular cell adhesion molecule 1(VCAM-1),phosphorylated nuclear transcription factorκB p65(p-NF-κB p65),phosphorylated nuclear factorκB inhibitory proteinα(pIκB-α)(by Western blot)and expression of interleukin-1beta(IL-1β),IL-6 and tumor necrosis factor-alpha(TNF-α)mRNA(using real-time polymerase chain reaction).Results Compared with group C,the LI,W/D ratio,Evans blue content in lung tissues and lung injury score were significantly increased,and the expression of IL-1βmRNA,TNF-αmRNA,IL-6 mRNA,VCAM-1,p-NF-κB p65 and pIKB-αwas up-regulated,and the expression of VE-cadherin was down-regulated in group LPS(P<0.05),and no significant change was found in the parameters mentioned above in group B(P>0.05).Compared with group B,the LI,W/D ratio,Evans blue content in lung tissues and lung injury score were significantly decreased,and the expression of IL-1βmRNA,TNF-αmRNA,IL-6 mRNA,VCAM-1,p-NF-κB p65 and pIKB-αwas down-regulated,and the expression of VE-cadherin was up-regulated in group LPS(P<0.05).Conclusions Bosutinib can ameliorate the acute lung injury in mice with endotoxemia.