乌司他丁对脓毒症小鼠肺损伤的改善作用及相关机制研究

Study on the effects of ulinastatin on lung injury in mice with sepsis and its related action mechanism

目的探讨乌司他丁对脓毒症小鼠肺损伤的改善作用及相关机制。方法选择成年SPF小鼠128只,随机分为对照组、假手术组、模型组和干预组。其中模型组和干预组使用CLP法建模。干预组尾椎静脉注射乌司他丁,其他2组注射等量0.9%氯化钠溶液。处理7 d后比较2组肺含水量、肺部组织血管通透性以及肺组织ROCK2及VE-cadherin表达情况。结果模型组肺组织W/D以及伊文思蓝含量显著高于假手术组和对照组(P<0.05),干预组W/D以及伊文思蓝含量显著低于模型组(P<0.05)。对照组和假手术组VE-cadherin主要表达于微血管内皮细胞,肺组织中ROCK2表达较少。模型组VE-cadherin表达阳性率显著低于假手术组和对照组,ROCK2显著高于假手术组和对照组(P<0.05)。干预组VE-cadherin表达阳性率显著高于模型组,ROCK2显著低于模型组(P<0.05)。模型组RhoA及ROCK1相对表达量均显著高于假手术组和对照组(P<0.05),干预组RhoA及ROCK1相对表达量显著低于模型组(P<0.05)。模型组血清NO显著低于假手术组和对照组(P<0.05),干预组血清NO显著高于模型组。结论乌司他丁具有改善脓毒症小鼠肺损伤的作用,可能是由于乌司他丁具有抑制Rho/ROCK信号通路,并通过VE-cadherin和NO调节肺毛细血管通透性的作用有关。

Objective To investigate the improvement effects of ulinastatin on lung injury in mice with sepsis,and to explore possible action mechanism.Methods A total of 128 adult SPF mice were randomly divided into control group, sham operation group, model group and intervention group. The animal models with sepsis wee established by CLP method in the mice of model group and the intervention group. The mice in intervention group were given ulinastatin by coccygeal vertebrae vein injection, and the mice in the other two groups were injected with equal volume of 0.9% sodium chloride solution. At 7 days after treatment, the lung water content, vascular permeability in lung tissue, and the expression levels of ROCK2 and VE-cadherin in lung tissue were observed and compared among groups.Results The W/D levels in lung tissues and Evans blue content in model group were significantly higher than those in sham operation group and control group(P<0.05). The W/D levels and Evans blue content in intervention group were significantly lower than those in model group(P<0.05). VE-cadherin was mainly expressed in microvascular endothelial cells in control group and sham operation group, and the ROCK2 expression was less in lung tissue. The positive rate of VE-cadherin expression in model group was significantly lower than that in sham operation group and control group, however,the ROCK2 expression was significantly higher than that in sham operation group and control group(P<0.05). The positive rate of VE-cadherin expression in intervention group was significantly higher than that in model group, but the ROCK2 expression was significantly lower than that in model group(P<0.05). The relative expression levels of RhoA and ROCK1 in model group were significantly higher than those in sham operation group and control group(P<0.05),moreover,the relative expression levels of RhoA and ROCK1 in intervention group were significantly lower than those in model group(P<0.05). In addition the serum levels of NO in model group were significantly lower than those in sham operation group and control group(P<0.05),which in intervention group were significantly higher than those in model group.Conclusion Ulinastatin has the effects of improving lung injury in mice with sepsis, which may be related with the inhibition effects of ulinastatin on Rho/ROCK signaling pathway, and the effects of VE-cadherin and NO in regulating pulmonary capillary permeability.