盐酸氨溴索对急性呼吸窘迫综合征大鼠肺纤维化的影响

Effect of ambroxol hydrochloride on pulmonary fibrosis in rats with acute respiratory distress syndrome

目的研究盐酸氨溴索对急性呼吸窘迫综合征(ARDS)大鼠肺纤维化的影响及作用机制。方法将60只雄性Wistar大鼠随机分为对照组(n=20)、模型组(n=20)及实验组(n=20)。模型组及实验组大鼠均用气管滴注3 mg·kg^-1脂多糖(LPS)100μL,构建ARDS大鼠模型。造模成功后,实验组大鼠腹腔注射盐酸氨溴索注射液40.0 mg·kg^-1·d^-1,对照组与模型组则注射等量生理盐水,连续干预21 d。用酶联免疫吸附法检测肺组织肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平及羟脯氨酸(HYP)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物(GSH-Px)含量,用蛋白质印迹法检测大鼠肺组织中核因子-κB(NF-κB)P65蛋白表达水平。结果对照组、模型组及实验组大鼠肺组织炎性因子TNF-α分别为(10.87±1.22),(36.93±3.14),(17.86±2.38)pg·g^-1,IL-6分别为(14.73±1.56),(37.12±4.61),(20.96±2.03)pg·g^-1,HYP分别为(319.55±46.37),(743.22±91.31),(415.38±57.88)μg·g^-1;MDA分别为(12.78±1.96),(33.45±5.16),(18.02±3.34)μmol·g^-1;SOD分别为(69.76±10.32),(28.63±2.89),(56.12±5.33)kU·g^-1,GSH-Px分别为(46.23±4.81),(15.04±2.39),(37.73±3.68)kU·g^-1,大鼠肺组织中NF-κB P65蛋白相对表达量分别为0.09±0.04,0.81±0.15,0.34±0.11。模型组和实验组与对照组比,以上各指标差异均有统计学意义(均P<0.05);实验组与模型组比较,差异均有统计学意义(均P<0.05)。结论盐酸氨溴索可降低炎症因子水平、减少氧自由基生成,抑制肺组织NF-κB P65蛋白表达,进而减轻ARDS大鼠肺损伤及肺纤维化。

Objective To investigate the effect and mechanism of amb-roxol hydrochloride on pulmonary fibrosis in rats with acute respiratory distress syndrome(ARDS). Methods Sixty SPF and male Wistar rats were randomly divided into control group(n=20), model group(n=20) and test group(n=20). Rats in model group and test group were treated with tracheal instillation of 3 mg·kg^-1 lipopolysaccharide(LPS) 100 μL, to construct the ARDS rat model. After successful modeling, the rats in test group were intraperitoneally injected with ambroxol hydrochloride 40.0 mg·kg^-1·d^-1, control group and model group were injected with the same amount of normal saline for 21 d.The levels of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), hydroxyproline ( HYP),malondialdehyde( MDA),superoxide dismutase( SOD) and glutathione peroxide( GSH-Px) content in lung tissue were detected by enzyme-linked immunosorbent assay;the expression of nuclear factor-κB( NF-κB)P65 protein in rat lung tissue was detected by Western blotting. Results The expression levels of TNF-α in lung tissue of control group,model group and test group were( 10. 87 ± 1. 22),( 36. 93 ± 3. 14) and( 17. 86 ± 2. 38)pg·g^-1,IL-6 were( 14. 73 ± 1. 56),( 37. 12 ± 4. 61),( 20. 96 ± 2. 03) pg·g^-1,HYP were( 319. 55 ± 46. 37),( 743. 22 ± 91. 31),( 415. 38 ± 57. 88) μg·g^-1,MDA were( 12. 78 ± 1. 96),( 33. 45 ± 5. 16),( 18. 02 ± 3. 34)μmol·g^-1,SOD were( 69. 76 ± 10. 32),( 28. 63 ± 2. 89),and( 56. 12 ± 5. 33) k U · g^-1,GSH-Px were( 46. 23 ± 4. 81),( 15. 04 ± 2. 39),( 37. 73 ± 3. 68) kU · g^-1,NF-κB P65 protein in rat lung tissues were0. 09 ± 0. 04,0. 81 ± 0. 15 and 0. 34 ± 0. 11. Compared with control group,above indexes in model group and test group were all had significant difference( all P < 0. 05),and there was significant difference between test group and model group( all P < 0. 05). Conclusion Ambroxol hydrochloride can reduce the level of inflammatory factors,reduce the production of oxygen free radicals,inhibit the expression of NF-κB P65 protein in lung tissue,and further reduce lung injury and pulmonary fibrosis in ARDS rats.