乌司他丁对水下爆炸继发性急性肺损伤兔基质金属蛋白酶-9等的影响

Effect of Ulinastatin on Matrix Metalloproteinase-9 and other cytokines in secondary acute lung injury in rabbits by underwater explosion

目的探讨乌司他丁对水下爆炸继发性急性肺损伤(ALI)兔基质金属蛋白酶-9(MMP-9)等细胞因子水平影响,探讨其治疗作用及机制。方法选择新西兰兔30只,随机数字表法分为3组:对照组(N组)、爆炸组(I组)和乌司他丁组(B组),每组10只,建立新西兰兔ALI动物模型后,B组经右颈外静脉滴注乌司他丁注射液(10万单位/kg),N组、I组注射等量生理盐水。12h采集血液,24h采集支气管肺泡灌洗液(BALF)、血液及肺组织标本,测定肺湿干重比(W/D),行12、24h血气分析,ELISA法测定BALF中性粒细胞弹性蛋白酶(NE)、血清中MMP-9、肿瘤坏死因子-α(TNF-α)、Western blot法和反转录PCR法测定肺组织MMP-9蛋白和mRNA的表达水平,光镜下观察肺HE切片病理学变化。结果 I组W/D、12h TNF-α、24h TNF-α、12h MMP-9、24h MMP-9均较N组升高[(5.79±1.60 vs.4.38±0.75)、(573.80±178.24 vs.348.12±172.13)ng/L、(552.30±169.64 vs.382.56±147.52)ng/L、(38.30±14.43 vs.23.67±5.28)ng/L、(42.10±15.24vs.25.72±7.16)ng/L,P<0.05],B组的W/D、12h TNF-α、24h TNF-α、24h NE、12h MMP-9、24h MMP-9较I组降低[(4.63±1.21 vs.5.79±1.60)、(356.40±181.23 vs.573.80±178.24)ng/L、(356.10±130.99 vs.552.30±169.64)ng/L、(111.90±10.81 vs.136.70±13.61)ng/L、(28.20±7.38 vs.38.30±14.43)ng/L、(30.50±8.07 vs.42.10±15.24)ng/L,P<0.05];肺病理切片显示B组肺水肿、炎症反应等较I组减轻。结论乌司他丁可抑制NMMP-9等细胞因子的表达,改善肺毛细血管通透性、减少渗出,减轻水下爆炸致家兔ALI的程度,可有效治疗ALI。

Objective To explore the effect of Ulinastatin（ UTI） on matrix metalloproteinase-9（ MMP-9）and other cytokines in secondary acute lung ingury（ ALI） in rabbits by underwater explosion. Methods Thirty rabbits were randomly divided into three groups： control group（ group N）,explosion group（ group I） and UTI group（ group B）. After establishing the models of ALI,the rabbits in group B were injected with UTI（ 10×104/kg） through right carotid vein,the other rabbits were injected with normal saline of same doses.The blood was collected at 12 h,and the bronchial alveolar lavage fluid（ BALF）,blood and some lung tissues were saved at 24 h. The content of Neutrophil elastase（ NE） in BALF,and tumor necrosis factor-alpha（ TNF-α） and MMP-9 in serum were detected by ELISA. The expression of MMP-9 in lung tissue was evaluated by Western blot and the reverse transcription polymerase chain reaction.The arterial blood gas was observed,the wet to dry weight ratios（ W/D） were determined,and the pathological changes of lung tissues were observed under the microscope. Results The levels of W/D,the TNF-α at 12 h,the TNF-α and NE at 24 h,and the MMP-9 at 12 h and 24 h were significantly higher in group I than those in group N[（ 5. 79 ± 1. 60 vs. 4. 38 ± 0. 75）,（ 573. 80 ± 178. 24 vs. 348. 12 ± 172. 13） ng/L,（ 552. 30 ± 169. 64 vs.382. 56±147. 52） ng/L,（ 38. 30±14. 43 vs. 23. 67±5. 28） ng/L,（ 42. 10±15. 24 vs. 25. 72±7. 16） ng/L（ P〈0. 05） ].The levels of W/D,the TNF-α at 12 h,the TNF-α and NE at 24 h,and the MMP-9 at 12 h and 24 h were significantly higher in group B than those in group I[（ 4. 63±1. 21 vs. 5. 79±1. 60）,（ 356. 40±181. 23 vs. 573. 80±178. 24） ng/L,（ 356. 10±130. 99 vs. 552. 30±169. 64） ng/L,（ 111. 90±10. 81 vs. 136. 70± 13. 61） ng/L,（ 28. 20± 7. 38 vs.38. 30±14. 43） ng/L,（ 30. 50±8. 07 vs. 42. 10±15. 24） ng/L（ P〈0. 05） ]. Histopathology of lung tissue showed that pulmonary edema and inflammatory were better in group B than those in group I. Conclusion UTI could restrain the expression of MMP-9 and other cytokines,improve pulmonary vasopermeability,relieve acute lung injury in rabbits by underwater explosion,and can effectively treat ALI.