氨溴索对百草枯中毒大鼠肺组织损伤NF-κB和细胞凋亡的影响

EFFECTS OF AMBROXOL ON NF-κB AND CELL APOPTOSIS IN LUNG INJURY ASSOCIATED WITH PARAQUAT POISONING OF RATS

目的探讨氨溴索(ambroxol,AT)对百草枯(paraquat,PQ)中毒大鼠急性肺损伤中性粒细胞(polymorphonuclearneutrophil,PMN)凋亡的影响。方法随机将108只大鼠分为4组,正常对照组(C组)、AT对照组(AT组)、急性肺损伤组(PQ组)和急性肺损伤+AT干预组(PQ+AT组)。取肺组织HE染色来评价肺组织损伤情况;ELISA检测血清中白细胞介素-8(interleukin-8,IL-8)水平;免疫细胞化学方法检测支气管肺泡灌洗液(bronchoalveolarlavage fluid,BALF)中PMN的核因子-κB(nuclear factor kappa B,NF-κB)p65表达;Western Blot分析定量检测PMN中胞浆蛋白IκBα的含量;流式细胞仪检测PMN凋亡率。结果 C组和AT组大鼠的肺组织结构基本完整;PQ组炎性细胞浸润明显增多;肺组织损伤程度加重。PQ+AT组肺组织损伤程度较轻。PQ组血清IL-8水平较C组显著增加(P<0.01),PQ+AT组较PQ组降低,较C、AT组高(P<0.01)。PQ中毒能够引起BALF中PMN的NF-κB亚基p65核易位和PMN胞浆蛋白IκBα的降解,AT对此具有抑制作用。PQ组3d时PMN的凋亡率最低,PQ+AT组相应时间点与PQ组相应时间点相比凋亡率均增加(P<0.05)。结论 PQ中毒导致了大鼠急性肺损伤,经AT治疗后,使延缓的PMN凋亡恢复正常,从而有效地减轻了PQ中毒所致的大鼠急性肺损伤。

Objective To investigate the effects of ambroxol （AT） on nuclear factor kappa B （NF- KB） and polymorphonuclear neutrophil （PMN） apoptosis in lung injury associated with paraquat （PQ） poisoning of rats. Methods A total of 108 rats were randomly divided into four groups： control group, group AT, group PQ and group PQ ＋ AT. Lung tissue was stained with hemotoxylin and eosin （HE） to evaluate the pathological change. The level of interleukin - 8 （ IL - 8 ） in serum was detected by ELISA. The expression of NF - KB p65 in PMN of bronchoalveolar lavage fluid （BALF） was determined by using immunohistochemiscal method. The content of IKBca of cytoplasm protein was determined by Western Blot analysis. The ratio of PMN apoptosis was assayed by flow cytometry. Results The structure of lung tissue was integrated in group C and group AT. Inflammatory cell infiltration increased obviously and the degree of lung injury was severe in group PQ ; The degree of lung injury was lighter in group PQ ＋ AT. The content of IL - 8 in group PQ was significantly higher than that of group C （ P 〈 0.01 ）. The content of IL - 8 in group PQ ＋ AT was lower than that of group PQ, and higher than that of group C andAT（ P 〈0. O1 ）. l）araquat poisoning caused NF- KB subunit p65 transh）cation and IKB（x degradation in PMN of BAI,F. Aml）roxol had inhibitoly effect on these changes. The ratio of PMN apoptosis in group PQ was the lowest on the third clay. The ratio of PMN apoptosis in group PQ ＋ AT was higher than that of group PQ （m corresponding time （ P 〈 O. 05） （ Conclusion Paraquat poisoning （：an lead to acute lung injury in rats. After using ambroxol （AT） ,the process of PMN apoptosis regain normal and lighten the lung injury associated with paraquat poisoning of rats.