Nuclear factor E2-related factor 2 attenuates endotoxin-induced acute lung injury by up-regulating cellular tight junction protein Claudin-18 expression
Objective To investigate the effect of nuclear factor E2-related factor 2(Nrf2)on the cellular tight junction protein Claudin-18 in endotoxin-induced acute lung injury(ALI).Methods Eighteen healthy male C57BL/6 mice were divided into control group,endotoxin-induced ALI model group(ALI group)and Nrf2 activator tert-butylhydroquinone(tBHQ)pretreatment group(tBHQ+ALI group)according to random number table method,with 6 mice in each group.Mice endotoxin-induced ALI model was reproduced by intraperitoneal injection of lipopolysaccharide(LPS,15 mg/kg),and the mice in the control group was injected with an equal amount of phosphate buffer solution(PBS).The mice in the tBHQ+ALI group received three intraperitoneal injections of tBHQ(a total of 50 mg/kg)at an interval of 1 hour before molding.The last injection of tBHQ was accompanied by LPS of 15 mg/kg.The mice in the control group and model group were given equal amounts of PBS,and PBS or LPS was given at the last injection.The mice were sacrificed at 12 hours after LPS injection to take lung tissues.After the lung tissue was stained with hematoxylin-eosin(HE)staining,the pathological changes were observed under light microscopy,and the lung injury score was calculated.The lung wet/dry ratio(W/D)was determined.Nrf2 protein expression in the lung tissue was detected by Western blotting.Positive expression of Claudin-18 in the lung tissue was determined by immunohistochemistry.Results The lung tissue showed normal structure,without significant pathological change in the control group.Compared with the control group,the alveolar septum widened accompanied by inflammatory cell infiltration,capillary hyperemia and tissue edema in the ALI group,the lung injury score and lung W/D ratio were significantly increased(lung injury score:6.50±1.05 vs.1.83±0.75,lung W/D ratio:3.79±0.22 vs.3.20±0.14,both P＜0.01),and the Nrf2 protein expression and Claudin-18 positive expression in the lung tissue were significantly lowered[Nrf2 protein(Nrf2/β-actin):0.41±0.33 vs.1.22±0.33,Claudin-18(A value):0.28±0.07 vs.0.44±0.10,both P＜0.05].After tBHQ pretreatment,the degree of lung histopathological injury was significantly reduced compared with the ALI group,the alveolar space slightly abnormal,inflammatory cell infiltration and tissue edema reduced,the lung injury score and lung W/D ratio were significantly decreased(lung injury score:3.00±0.89 vs.6.50±1.05,lung W/D ratio:3.28±0.19 vs.3.79±0.22,both P＜0.01),and Nrf2 protein expression and Claudin-18 positive expression in the lung tissue were significantly increased[Nrf2 protein(Nrf2/β-actin):1.26±0.09 vs.0.41±0.33,Claudin-18(A valure):0.45±0.04 vs.0.28±0.07,both P＜0.05].Conclusion Nrf2 alleviated pulmonary edema and improved endotoxin-induced ALI by up-regulating Claudin-18 expression.
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